Journal of Shandong University (Health Sciences)

Immunomodulatory effects of lipopolysaccharide-induced Treg imbalance on experimental autoimmune myasthenia gravis

LI Liangkang, SI Weiyue, WU Xingyuan, ZHOU Yang, WEI Shuli, DONG Jing, LI Xiaoli, DUAN Ruisheng

Journal of Shandong University (Health Sciences). 2023, 61(10): 1-8. doi:10.6040/j.issn.1671-7554.0.2023.0196

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Objective To investigate the changes and functional characteristics of immune cells in experimental autoimmune myasthenia gravis(EAMG)complicated with acute infection. Methods Female Lewis rats aged 6-8 weeks were immunized with R97-116 peptide of rat acetylcholine receptor alpha subunit. Lipopolysaccharide(LPS)was injected intraperitoneally to induce acute infection(LPS group), and PBS was injected intraperitoneally to set controls(PBS group). Clinical scores and body weight were recorded. Flow cytometry was used to detect the frequencies of Th1, Th17 and Treg cells from lymphoid organs and peripheral circulation, and follicular helper T(Tfh)cells, dendritic cells(DCs)and germinal center B(GCB)cells in the spleen. Peanut agglutinin(PNA), CD4 and Foxp3 levels in the spleen were detected with immunofluorescence. The levels of anti-R97-116 IgG antibody and its subclasses in serum were analyzed with ELISA. Results Compared with the PBS group, the LPS group had aggravated clinical symptoms and lower body weight. The frequency of Treg(CD4⁺CD25⁺)in the spleen was decreased(P=0.016), and the Foxp3 expression in the spleen was reduced. In addition, the PNA in the spleen of the LPS group was increased and clustered, and the expression of MHC-Ⅱ(P=0.002)and secretion of IL-6(P=0.017)of GCB cells were increased. The mean fluorescence intensity(MFI)of DC subclass expressing CD80 was increased but not statistically significant(P=0.057). The level of protective antibody IgG1 in peripheral blood was decreased in LPS group(P=0.005), the level of pathogenic antibody IgG2b did not change, and IgG1/IgG2b was down-regulated(P=0.018). Conclusion LPS may reduce the frequency of Treg cells and the expression of Foxp3 in the spleen, enhance the germinal center response, function of GCB cells and ability of DC co-stimulation, eventually leads to the aggravation of clinical symptoms of EAMG rats.

Effects of Luling Anshen Granule on the inflammatory response and cell apoptosis in rats with chronic renal failure

YANG Yuanfeng, XIONG Gaocai, LI Yuchuan, LUO Yuling, ZHANG Jingjie

Journal of Shandong University (Health Sciences). 2023, 61(10): 9-16. doi:10.6040/j.issn.1671-7554.0.2023.0050

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Objective To investigate the effects of Luling Anshen Granule on adenine-induced chronic renal failure(CRF)rats and its potential mechanism. Methods Male SD rats were divided into 6 groups: control group, model group, positive drug group(Niaoduqing Granule, 2.25 g/kg), low-, medium- and high-dose Luling Anshen Granule groups(2.5, 5, 10 g/kg), with 10 rats in each group. Except the control group, the other groups were given 2.5% adenine suspension by gavage to establish CRF models, and corresponding drugs were given by gavage once a day for a consecutive of 28 days after successful modeling. The general condition of rats was observed and recorded. Blood was sampled from the abdominal aorta to measure the levels of blood urea nitrogen(BUN)and serum creatinine(Scr), and the level of 24-hour urinary protein was also measured. The levels of interleukin-1β(IL-1β), interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)in serum were determined with ELISA. Histopathological changes of kidney tissues were observed with HE staining. The level of basic fibroblast growth factor(bFGF)in renal tissues was detected with immunohistochemistry. The mRNA expressions of Bax, Bcl-2 and Caspase-3, and the protein expressions of Bax, Bcl-2 and Cleaved-Caspase-3 in renal tissues were detected with qRT-PCR or Western blotting. Results Compared with the control group, the model group had more serious renal injury, and significantly increased levels of 24-hour urinary protein, BUN, Scr, IL-1β, IL-6, TNF-α, mRNA expressions of Bax/Bcl-2 and Caspase-3, protein expressions of Bax/Bcl-2 and Cleaved-Caspase-3, and bFGF(P<0.05). Compared with the model group, the Luling Anshen Granule intervention groups and positive drug group had significantly alleviated renal injury, and significantly decreased levels of the above indexes in a dose-dependent manner(P<0.05). Conclusion Luling Anshen Granule can inhibit the inflammatory response, cell apoptosis and fibrosis in kidney tissues of CRF rats and improve the renal function, thus having a protective effect on the kidney.

Mechanism of umbilical cord mesenchymal stem cells treating bleomycin-induced pulmonary interstitial fibrosis in mice

WANG Cheng, ZHANG Shuhui, YAN Na, MENG Linlin, CHEN Liangzhi, LIU Qinghua

Journal of Shandong University (Health Sciences). 2023, 61(10): 17-22. doi:10.6040/j.issn.1671-7554.0.2023.0092

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Objective To explore the therapeutic effects of umbilical cord mesenchymal stem cells(MSCs)on advanced pulmonary fibrosis. Methods A total of 39 male Kunming mice were randomly divided into 3 groups: normal control group(n=12, NC group), bleomycin group(n=12, PC group)and MSCs treatment group(n=15, MSCs group). On day 1, bleomycin was injected into the trachea of mice in the PC and MSCs groups, and the same amount of saline was injected into mice in the NC group. On day 21, 3 mice in the MSCs group were dissected and the other mice were injected with MSCs into the tail vein. On day 42, all mice were sacrificed. The lung wet weight, hydroxyproline and the expression of transforming growth factor beta(TGF-β)and interferon-gamma(IFN-γ)were compared among the 3 groups. Results On day 21, the alveolar texture was disordered in the MSCs group, the alveolar interval was widened, and the pulmonary interstitial fibrosis was formed. On day 42, the alveolar texture was visible in the NC group and there was no alveolar interval widening; the alveolar texture was disordered in the PC group and the interval was significantly widened; the alveolar texture was clear in the MSCs group, and the alveolar interval was widened. The PC group had significantly higher lung wet weight, hydroxyproline content and TGF-β expression than the NC group(P<0.01)and MSCs group(P<0.05). The MSCs group had significantly higher IFN-γ expression than the PC group(P<0.01), followed by the NC group(P<0.01). Conclusion MSCs can protect lung tissue, and inhibit and alleviate the formation of intermediate and late pulmonary interstitial fibrosis in mice.

Constructing and validating a prognostic model for patients with stage IIIB non-small cell lung cancer based on SEER database

ZHAO Qidi, WANG Kai, ZHAO Xiaogang, YAN Tao, WANG Yadong, DU Jiajun

Journal of Shandong University (Health Sciences). 2023, 61(10): 23-37. doi:10.6040/j.issn.1671-7554.0.2022.0743

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Objective To investigate the prognosis of patients with stage IIIB non-small cell lung cancer(NSCLC)and to construct a prognostic tool that can be widely used in clinical practice. Methods Clinical data of patients with stage IIIB NSCLC were retrospectively collected from the Surveillance, Epidemiology and End Result(SEER)databases, and randomly grouped into the training set and validation set with a 7∶3 ratio. Information of patients with stage IIIB NSCLC treated in the Department of Thoracic Surgery of Shandong Provincial Hospital Affiliated to Shandong University during 2005 and 2019 was collected as a secondary validation set(SPH validation set). Univariate and multivariate Cox proportional hazards regression analyses were conducted in the training set to construct a nomogram and web tool. Receiver operating characteristic(ROC)curve, concordance index(C-index), and decision curve analyses were performed using R software to evaluate the nomogram and assess the clinical benefits. Results A total of 7,903 patients were involved, including 5,532 in the training set and 2,371 in the validation set. Altogether 10 variables were included in the nomogram, including gender, age, primary site of tumor, grade of tumor, histology of tumor, tumor-node-metastasis stage specific subgroups of ⅢB stage in the eighth edition, dissection of regional lymph node, extent of surgery, radiotherapy and chemotherapy. The area under the ROC curve(AUC)of 5-year overall survival(OS)was 0.724 and 0.708 in the training and validation sets, respectively. The C-index was 0.661, 0.649 and 0.685 in the training, validation and SPH validation sets, respectively. The calibration plot showed that the model had good predictive power. Surgery with chemotherapy was identified as the optimal treatment strategy compared to chemotherapy or radiotherapy alone(P<0.001). An interactive web tool was also constructed. Conclusion A prognostic nomogram was successfully constructed and an interactive web tool to predict patients’ survival was built.

Effects of sodium bicarbonate on the visual clarity and disease detection rate of gastroscopy

LI Ran, GUO Anbing, ZHANG Xiuyun, LU Lin, DU Chao

Journal of Shandong University (Health Sciences). 2023, 61(10): 38-45. doi:10.6040/j.issn.1671-7554.0.2023.0206

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Objective To verify that the administration of sodium bicarbonate solution before gastroscopy can have comparable defoaming effects as the commonly used defoaming medications in clinical practice without affecting the detection rate of endoscopic lesions. Methods In this double-blind, single-center, randomized controlled trial, 950 patients undergoing gastroscopy were involved, who were randomly assigned to 6 groups. A, B, and C were experimental groups, while D, E and F were control groups. In group A, 1 g of sodium bicarbonate was dissolved in 50 mL of warm water at 40 ℃; in group B, 2 g of sodium bicarbonate was dissolved in 50 mL of warm water at 40 ℃; in group C, 1 g of sodium bicarbonate+3 mL of simethicone was dissolved in 50 mL of warm water at 40 ℃; in group D, 20 000 U of pronase(containing 1g of sodium bicarbonate)+3 mL of simethicone was dissolved in 50 mL of warm water at 40 ℃(the fixed gastroscopy premedication in our center); in group E, 1.2 g of N-acetylcysteine(NAC)+3 mL of simethicone was dissolved in 50 mL of warm water at 40 ℃; in group F, 50 mL of warm water at 40 ℃ was used. All patients routinely drank 10 mL of lidocaine gastroscopic lubricant to anesthetize the pharynx 5 min before the procedure. After completion of the examination, the operating physicians filled in a rating form for the mucosal visibility score(MVS)and operational satisfaction score(OSS), recorded the endoscopic diagnosis, and calculated the total mucosal visibility score(TMVS). Results Group F was the least effective in removing blisters; only 2% of the patients had TMVS of 5 and only 71% achieved physician satisfaction, significantly lower than those of the other 5 groups; the differences in scores for each site and physician satisfaction were statistically significant compared to the other 5 groups(P<0.003). Group D was the most effective in removing blisters; 70% of the patients had TMVS of 5, and 98% achieved physician satisfaction. Groups A and D had comparable effects in removing blisters; there was only difference in TMVS(P<0.003). There was only significant difference in MVS of gastric fundus between group A and group C(P<0.003). There were differences in the MVS of gastric fundus, gastric body and TMVS between group C and group D(P<0.003). The overall efficacy of defoaming in Group C was average. Between group A and group B, the differences in all scores were statistically significant(P<0.003), and the efficacy of foam removal was significantly better in group A. No statistically significant differences in all scores were observed among groups E, A and C, and the efficacy of foam removal was approximately the same. There was no significant difference in the detection rate of endoscopic lesions among the 6 groups(P>0.003). Conclusion Administration of 1 g of sodium bicarbonate solution before gastroscopy can achieve approximately the same efficacy as the commonly used defoaming medications without affecting the detection rate of endoscopic lesions. The administration of 1 g is more effective than 2 g, and the addition of simethicone may not yield better results. Sodium bicarbonate is inexpensive and worth promotion in primary hospitals.

Efficacy of nonsynchronous CT-assisted localization combined with ultrasound localized minimally invasive percutaneous nephrolithotomy under local anesthesia for upper urinary calculi

ZHANG Yingchen, ZHAO Shuai, SUN Hao, BI Wenhao, WU Zhiqiang, CHENG Xiaojian, JIANG Licheng

Journal of Shandong University (Health Sciences). 2023, 61(10): 46-50. doi:10.6040/j.issn.1671-7554.0.2023.0827

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Objective To investigate the value of nonsynchronous CT-assisted localization combined with ultrasound localized minimally invasive percutaneous nephrolithotomy(MPCNL)under local anesthesia in the treatment of upper urinary calculi. Methods A total of 259 sides in 244 cases of upper urinary calculi underwent CT scan localization to simulate intraoperative posture before operation, and MPCNL was then successfully performed under local anesthesia. Visual analog scale(VAS), stone-free rate and complications were recorded. Results All cases were operated under satisfactory local anesthesia. The VAS was 1-3 in 112 cases(45.9%), 4-6 in 117 cases(48%), and 7-9 in 15 cases(6.1%). Percutaneous puncture channels were successfully established in all patients. The operation time was 40-160 min, with an average of 92 min. The stone-free rate reached 88.8%(230/259)in all cases, 98.1%(102/104)in the upper ureteral calculi group, 93.9%(62/66)in the single renal calculi group, and 74.2%(66/89)in the complex renal calculi group. After operation, active bleeding occurred in 2 cases. No severe complications such as injury of liver, spleen, thorax or intestines were observed. The length of hospital stay was 5-15 d, with an average of 8.6 d. Conclusion Nonsynchronous CT-assisted localization combined with ultrasound localized MPCNL under local anesthesia is simple, safe, economical and effective, which justifies promotion in clinical practice.

Clinical features of dentatorubral pallidoluysian atrophy

KONG Liangbo, XIA Ruihong, ZHAO Yuying, WANG Shuzhen, WANG Shengjun

Journal of Shandong University (Health Sciences). 2023, 61(10): 51-57. doi:10.6040/j.issn.1671-7554.0.2023.0177

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Objective To analyze the clinical manifestations, gene changes and brain MRI in patients with dentatorubral pallidoluysian atrophy(DRPLA). Methods The clinical characteristics of 3 DRPLA patients treated in our hospital were collected, and the clinical data of DRPLA patients reported in our country were summarized. Results Clinical data from a total of 35 DRPLA patients were collected. The average age of onset was(25.5±15.4)years, and the range of cytosine-adenine-guanine(CAG)repetitions at 12p13.31 of ATN1 gene was 53-79 times. The age of onset was negatively correlated with the number of CAG repetitions(r=-0.846, P<0.001). The main clinical manifestations of different subtypes, including juvenile type, early-onset adult type, and late-onset adult type, were quite different. Juvenile patients usually had seizures, cognitive impairment and walking instability, complicated with involuntary movements or mental disorder; Early-onset adult patients typically had walking instability, involuntary movement and cognitive impairment, with approximately half of the patients exhibiting seizures; Late-onset adult type patients had signs of walking instability, but seizures were rare. Brain MRI of 29 patients showed that cerebellar atrophy(93.1%, 27/29)and brainstem atrophy(69.0%, 20/29)were the most frequently observed abnormalities, followed by cortical atrophy(51.7%, 15/29)and white matter lesions(51.7%, 15/29). Conclusion High number of repeats of CAG sequence of ATN1 gene in DRPLA patients is associated with young age of onset. The clinical manifestations are diverse. For patients with epilepsy, ataxia, cognitive impairment or involuntary movement, along with a positive autosomal dominant family history, both brain MRI and ATN1 gene detection are recommended.

A Mendelian randomization study of aspirin use and the risk of endometrial cancer

CHANG Xin, LIU Shijia, HAN Lu

Journal of Shandong University (Health Sciences). 2023, 61(10): 58-62. doi:10.6040/j.issn.1671-7554.0.2023.0451

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Objective To investigate the causal relationship between aspirin use and the risk of endometrial cancer using the two-sample Mendelian randomization method. Methods A two-sample Mendelian randomization analysis was performed to estimate the potential causal relationship between aspirin use and endometrial endometrioid carcinoma(EEC)and non-endometrioid carcinoma(NEC)using GWAS genome-wide data. The single nucleotide polymorphism(SNP)data associated with aspirin use, EEC and NEC were obtained from the GWAS catalog database. SNPs with strong association with aspirin use were selected as instrumental variables and analyzed with inverse variance weighted(IVW), weighted median, MR Egger, simple mode and weighted mode. The causal relationship between aspirin use and risk of EEC and NEC was evaluated with odds ratio(OR). Results A total of 6 SNPs were included as instrumental variables, and the IVW showed that aspirin administration reduced the incidence of EEC(OR=0.02, 95%CI: 0-0.28, P=0.005), while no causal relationship was found between aspirin use and the risk of NEC(OR=0.00, 95%CI: 0.00-16.26, P=0.199). The results of other methods were consistent with those shown by IVW. Sensitivity analysis indicated that heterogeneity, horizontal pleiotropy and leave-one-out tests showed reliability of instrumental variables in different tissue types of endometrial cancer with no significant bias in the results. Conclusion Aspirin can reduce the risk of EEC. Exploring the biological mechanism of aspirin on EEC at a deeper level has important implications for the early prevention and treatment of endometrial cancer.

Effects of GnRH-a pretreatment on the pregnancy outcomes of frozen embryo transplantation in patients with endometriosis

ZHANG Lixia, TIAN Yizheng, LYU Shangge, ZHANG Yanan, YAN Lei

Journal of Shandong University (Health Sciences). 2023, 61(10): 63-68. doi:10.6040/j.issn.1671-7554.0.2023.0500

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Objective To investigate the effects of pretreatment of gonadotrophin releasing hormone agonist(GnRH-a)for hormone replacement treatment(HRT)on the pregnancy outcomes in patients with endometriosis(EMs)after frozen embryo transplantation(FET). Methods Clinical data of 643 EMs patients who received FET in the Affiliated Reproductive Hospital of Shandong University during Jan. 2016 and Dec. 2019 were retrospectively analyzed. According to the endometrial preparation regimen, the patients were divided into experimental group(GnRH-a+HRT group, 261 frozen embryo transfer cycles)and control group(HRT group, 382 frozen embryo transfer cycles). The pregnancy outcomes were compared between the two groups, including live birth rate and abortion rate. Binary Logistic regression was used to correct confounding factors affecting pregnancy outcomes. Results There were no significant differences in the number of days of basic luteinizing hormone(LH), basic estrogen(E2), basic prolectin(PRL), basic thyrotropin(TSH), gonadotropin(Gn)and infertility types between the two groups(P>0.05). The age, basic follicle stimulating hormone(FSH), total amount of Gn and daily intimal thickness of human chorionic gonadotropin(HCG)were higher in the GnRH-a+HRT group, while anti-Mullerian hormone(AMH), antral follicle count(AFC), body mass index(BMI)and basic androgen(T0)were lower in the HRT group(P<0.05). After adjusting for confounders, the GnRH-a+HRT group was associated with a lower risk of miscarriage(OR=0.607, 95%CI=0.378-0.973, P=0.038)but not with live birth rate. Old age(OR=0.931, 95%CI=0.891-0.973, P=0.002)and high FSH level(OR=0.924, 95%CI=0.855-0.998, P=0.043)were negatively associated with live birth rate. Conclusion GnRH-a pretreatment for endometriosis patients undergoing FET before HRT can reduce the risk of abortion, but cannot significantly improve the live birth rate. Further study is needed to confirm the conclusion.

Relationship between preoperative cartilage condition and postoperative VAS in patients undergoing high tibial osteotomy

LI Lei, LU Qunshan, ZHANG Baoqing, HAN Kaifei, ZHENG Tong, LI Binglong, LIU Peilai

Journal of Shandong University (Health Sciences). 2023, 61(10): 69-73. doi:10.6040/j.issn.1671-7554.0.2023.0144

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Objective To evaluate the relationship between preoperative cartilage injury and clinical efficacy of high tibial osteotomy(HTO)by analyzing the postoperative visual analogue scale(VAS)of knee pain. Methods Clinical data of 42 patients with knee osteoarthritis treated with surgery and followed up during Oct. 2018 and May 2021 were retrospectively analyzed. The patients were divided into Outbridge Ⅲ group and Outbridge Ⅳ group. The operation time, intraoperative blood loss, intraoperative cartilage injury under knee arthroscopy, postoperative drainage volume, and follow-up results at 6 weeks, 3 months, 6 months, and 1 year were statistically analyzed. The degree of knee pain was evaluated with VAS scoring system. Results All 42 patients were followed up for an average of 7.39 months(6-12 months). All osteotomy fracture lines were clinically healed. There were no significant differences in operation time, postoperative drainage volume, intraoperative blood loss and VAS between the two groups(P>0.05). Conclusion Patients with Outbridge Ⅲ and Outbridge Ⅳ knee osteoarthritis can obtain satisfactory clinical results after HTO.

Establishment and evaluation of a recombinase-aided amplification for the detection of human adenovirus type 4

LIU Qingqing, WANG Ningning, CHENG Jun, ZHOU Huajun, CHE Feihu, SUN Qingyang, WANG Yue, ZHANG Yingjie, DAI Yuzhu

Journal of Shandong University (Health Sciences). 2023, 61(10): 74-82. doi:10.6040/j.issn.1671-7554.0.2023.0640

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Objective To establish a simple and rapid method for human adenovirus type 4(HAdV-4)detection with recombinase-aided amplification(RAA). Methods The specific primers and probes were designed based on the Hexon gene of HAdV-4. The constructed positive plasmid standard was used for RAA under different temperatures(39 ℃, 40 ℃, 42 ℃)to select the best reaction temperature, and the sensitivity, repeatability, and specificity of the method were tested. A total of 100 clinical samples were detected with real-time RAA, and the results were compared with those of real-time PCR to evaluate the detection performance of real-time RAA. Results The optimal temperature for real-time RAA was 42 ℃, and the detection limit was 10¹ copies/μL. When the concentration of the plasmid standard was 10¹-10⁴ copies/μL, the intra-assay coefficient of variation was less than 5%, showing good repeatability. The results of specificity showed that the method accurately detected HAdV-4 without cross-reactivity with other subtypes or other pathogens. Real-time PCR and real-time RAA of the 100 samples showed the coincidence rate was 100%. Conclusion A specific, simple and rapid assay for HAdV-4 detection is developed.

Glove perforation and its influencing factors in 141 cases of subgingival scaling

JIANG Lili, CHEN Peng, XU Xinyang, ZHANG Xiaorong, MENG Qiuyu, QI Dong, YANG Chuanjing, WU Qian

Journal of Shandong University (Health Sciences). 2023, 61(10): 83-87. doi:10.6040/j.issn.1671-7554.0.2023.0278

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Objective To investigate incidence of perforation of gloves occurred in subgingival scaling and to explore the influencing factors. Methods Gloves used in subgingival scaling were collected in the Department of Periodontology of a stomatological hospital from July 2021 to February 2022, and glove perforation was tested with the impermeability test. A general information questionnaire was used to collect information of glove users. SPSS 25.0 statistical software was used for statistical description. Chi-square test, independent-sample t-test or Fisher exact probability method and Mann-Whiney U test were used to analyze glove perforation and the influencing factors. Results A total of 25 cases of perforation occurred in 141 pairs of gloves, and the incidence of perforations was 14.9%(21/141). Glove perforation occurred mostly in the right-hand, accounting for 64.0%(16/25), especially on the right index finger(6 cases)and right palm(4 cases). There was significnat difference in the operation time(U=767.500, P=0.004). Conclusion The incidence of glove perforation in subgingival scaling is high, and most perforation sites are the right index finger and palm. Operation time is a significant influencing factor of glove perforation.

Outcome and influencing factors of 314 outpatient smoking cessation cases

QIU Yifan, WANG Zihan, GAO Tingting, WANG Xiyan, SUN Yan, LIU Fen, QI Qian, DONG Liang

Journal of Shandong University (Health Sciences). 2023, 61(10): 88-94. doi:10.6040/j.issn.1671-7554.0.2023.0202

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Objective To compare the effects of drug quitting and psychological behavior intervention on smoking cessation, and to compare the success rate of smoking cessation after 1, 3 and 6 months of follow-up in the real world. Methods During Jun. 1, 2022 and Dec. 31, 2022, 314 smoking cessation patients registered in the Smoking Cessation Clinic of Shandong Provincial Qianfoshan Hospital were divided into two groups according to different ways of intervention. The medication-assisted group included 178 smokers who chose to use drugs(113 used Bupropion, 38 used Varenicline and 27 used Nicotine patches), while the control group included 136 patients who accepted the psychological behavioral intervention. The smoking cessation rate of the two groups after 1 month, 3 months and 6 months were analyzed with SPSS 26.0 software to identify the differences in the effects of interventions and the influencing factors. Results The success rate of drug-assisted quitters was 48.3%, the success rate of psychological behavioral quitters was 13.2%, with statistically significant difference between the two groups(χ2=51.714, P<0.05). Smokers with a bachelors degree or above and a daily smoking limit of less than 1 box(20 cigarettes)were more likely to quit smoking, which were protective factors for smoking cessation success. Conclusion Drug-assisted smoking cessation and psychological behavior intervention are both good ways to assist smoking cessation. The former is more helpful, especially in reducing the rate of relapse.

Molecular genetic mechanism of Duchenne muscular dystrophy in a female case

WU Wenjing, SUN Yuan, WANG Guangyu, LYU Xiaoqing, YAN Chuanzhu, LIN Pengfei

Journal of Shandong University (Health Sciences). 2023, 61(10): 95-100. doi:10.6040/j.issn.1671-7554.0.2023.0199

Abstract ( 201 )

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Objective To explore the clinical phenotypes and gene variation characteristics of a female child with Duchenne muscular dystrophy, and to explore its molecular genetic pathogenesis. Methods Clinical data and muscle biopsy of the child were analyzed. Multiplex ligation-dependent probe amplification(MLPA)and Ion Torrent amplicon sequencing were performed to detect variants in the DMD gene of the child and her parents. Karyotype analysis and array-based comparative genomic hybridization(a-CGH)were carried out for the detection of chromosome variations. In addition, human androgen receptor gene methylation-specific PCR(HUMARA-MSP)was conducted to identify the X chromosome inactivation skewing. Results The patient was three years old at the first visit. The main clinical symptoms were increased creatine kinase(CK)level and reduced athletic capacity. Muscle pathology showed myodystrophic changes and loss of dystrophin protein. MLPA did not detect deletion and duplication mutations in the DMD gene. Ion Torrent amplicon sequencing revealed heterozygous c.10223 + 1G>A in the DMD gene, a spontaneous mutation. Chromosome karyotypes were normal, and a-CGH did not detect chromosomal abnormalities. Significant X chromosome inactivation skewing in this patient was confirmed by HUMARA-MSP. Conclusion The child carried a heterozygous c.10223+1G>A in the DMD gene, and the X chromosome inactivation skewing was the cause of DMD.

Causal mediation analysis with multiple-mediator of hypertension and depression between type 2 diabetes mellitus and cognitive function based on G-computation

HAN Mei, MENG Weijing, TAO Zikun, YANG Xi, XU Yaqi, MU Huaxia, BU Weixiao, WANG Suzhen, SHI Fuyan

Journal of Shandong University (Health Sciences). 2023, 61(10): 101-108. doi:10.6040/j.issn.1671-7554.0.2023.0647

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Objective To explore the direct causal effect between type 2 diabetes mellitus(T2DM)and cognitive function, as well as the mediating effect of hypertension and depression in the causal relationship. Methods A total of 1 306 participants aged 60 to 80 years were selected from the National Health and Nutrition Examination Survey(NHANES)database for the years 2013-2014 and divided into the control group and T2DM group according to whether they had T2DM. The mediating effects of binary mediator hypertension and continuous mediator depression on the relationship between T2DM and cognitive function was analyzed with G-computation causal multi-mediation, and subgroup analysis was performed according to genders. Results There was a statistically significant direct effect between T2DM and Animal Verbal Fluency Test and Digit Symbol Substitution Test, with b and 95%CI of -0.75(95%CI: -1.45 - 0.04)and -4.10(95%CI: -6.02 - 2.18), respectively. Hypertension and depression played a partial mediating role in the relationship between T2DM and cognitive function, and b and 95%CI were -0.11(95%CI: -0.21-0.00)and -0.50(95%CI: -0.94 - 0.05), respectively. Subgroup analysis showed that the direct and indirect effects were statistically different between genders. Conclusion The cognitive decline induced by T2DM may be mediated by hypertension and depression, and gender may be a moderating variable. Prevention, management and treatment of the complications play an important role in reducing the risk of cognitive dysfunction.

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