Objective:To observe effects of wrist-ankle acupuncture (WAA) onβ-endorphin (EP), nitric oxide (NO) in uterus tissue and prostaglandin F2α (PGF2α), substance P (SP) in serum of rats with primary dysmenorrhea. Methods:A total of 45 non-pregnant Wistar rats were randomly divided into a control group, a model group and a WAA group, 15 rats in each group. Rats in the model group and the WAA group received continuous abdominal subcutaneous injection of Diethylstilbestrol to establish dysmenorrhea rat models. On the first day after modeling, rats in the WAA group began to receive acupuncture on Point Lower 1 and Point Lower 2, once a day for 10 d. The control group and the model group didn’t receive any treatment. Writhing latencies and frequencies were recorded.β-EP and NO in uterus tissue homogenates and PGF2α, SP in serum were detected. Results:In the model group,β-EP and NO levels were the lowest among the groups, the serum PGF2α level was the highest, and serum SP level was the lowest. These measurements showed significantly difference between the model group and the control group (P<0.05). PGF2α in the WAA group was lower than that in the model group;β-EP, NO and SP levels were higher than those in the model group, with inter-group statistically significant differences (P<0.05). Conclusion: WAA may achieve analgesic effect through decreasing PGF2α, increasingβ-EP, NO and SP to relieve uterine cramps, increase blood flow and promote functional improvement.

ObjectiveTo observe the therapeutic efficacy of warm needling plus acupoint injection in treating post-stroke urinary retention.MethodSeventy eligible subjects with post-stroke urinary retention were randomized into a treatment group of 35 cases and a control group of 35 cases. The treatment group was intervened by warm needling plus acupoint injection, while the control group was by warm needling alone. The relevant symptoms of urinary retention were observed before and after treatment.Result After 1-week treatment, the retention condition was improved in both groups, and the treatment group was significantly superior to the control group (P<0.05); The 2-week and 1-month follow-up studies showed that the retention condition in the treatment group was significantly better than that in the control group (P<0.05). The adverse events in the treatment group were significantly less than that in thecontrol group (P<0.05).ConclusionWarm needling plus acupoint injection can produce a satisfactory efficacy in treating post-stroke urinary retention and is worth application in clinic. However, this method still needs improving as its effect on initialurination and urination duration is not significant.

Objective To compare the relationship between the enzyme‐linked immunosorbent assay(ELISA) reagent and West‐ern blot(WB) confirmation reagent for analyzing the quality lever of human T‐cell lymphotropic virus(HTLV) detection reagent . Methods The WB confirmation reagent was used to detect anti‐HTLV antibody in 156 human serum samples of ELISA prelimina‐ry screening positive .The ELISA cut‐off value(optimal value) was selected by using the two‐graph receiver operating characteristics (TG‐ROC) analytical method .The two‐by‐two table analysis was constructed to analyze the consistency of results detected by the two methods ,moreover the McNemar test was used to evaluate the consistency of detection results .The quality level of HTLV de‐tection reagent was comprehensively evaluated .Results Among 156 serum samples of ELISA preliminary screening positive ,only 40 samples were positive by the WB confirmation ,and other 116 samples were negative .The sensitivity and specificity of ELISA de‐tection reagent obtained by TG‐ROC analysis were 97 .5% and 45 .7% respectively ,the TG‐ROC test also indicated that the detec‐tion results had significant difference between ELISA and WB(P<0 .05) .By adjusting the cut‐off value ,the sensitivity and specific‐ity of ELISA were increased to 88 .8% (parametric method) .In the comparison of the parametric method and the non‐parametric method ,the obtained areas under the curve(AUC) was 0 .923 5(parametric method) ,their results were basically consistent .Conclu‐sion Although above results indicate that the detection results of ELISA reagent are different from those of WB ,but adjusting the cut off value can increase its sensitivity and specificity ,thus increases the reliability of diagnosis result .

Objective To explore the relationship between cerebral micro-bleeds (CMBs) and recurrent stroke after transient ischemic attack (TIA),and to guide early diagnosis of prognoses.Methods A total of 147 patients with TIA,admitted to our hospital from January 2015 to June 2017,were enrolled;29 patients (19.73％) were given anti-platelet therapy,double-antibody treatment was performed in 103 patients (70.07％),15 patients (10.20％) were given anticoagulation,and 108 patients (73.47％) took statins.Intracranial imaging was performed on these patients on admission and one week after different treatments.The general data and biochemical markers were collected;the recurrent stroke after TIA within 90 d was followed up.Univariate analysis and multivariate Logistic regression were used to screen the risk factors for stroke after TIA.Results CMBs were noted in 30 patients (20.41％) at first intracranial imaging;another 2 patients with CMBs were noted at one week after treatments,and there were no significant differences in newly-detected CMBs among different treatment methods (P＞0.05).Nine patients (6.12％) had stroke recurrence.Univariate analysis indicated that the percentages of patients with atrial fibrillation,symptomatic carotid artery stenosis≥ 50％,CMBs and white matter high signal (WMH) and high ABCD2 scores between TIA patients with/without stroke were significantly different (P＜0.05).Multivariate Logistic regression showed that CMBs were the independent risk factors of recurrent stroke after TIA (OR=4.126,P=0.003,95％CI:0.320-2.390).Conclusion CMBs canpredict the risk of stroke recurrence after TIA.

Objective: To evaluate whether simultaneous vaccination with live attenuated polio vaccine affects the immunogenicity of live attenuated rotavirus (RV) vaccine. Methods: Rotarix produced by GlaxoSmithKline was used as the research object. Two doses of Rotarix were orally administered on day 0 and month 1, and oral live attenuated polio vaccine (OPV) was administered on day 0, month 1 and month 2 according to the national vaccination plan. Healthy infants aged 6 to 16 weeks were randomly divided into two groups: interval vaccination group (Rotarix and OPV were vaccinated on different days) and simultaneous vaccination group (Rotarix and OPV were vaccinated on the same day). Serum samples were collected on day 0, month 2 and month 12, and serum RV-IgA was measured by enzyme linked immunosorbent assay. Statistical analysis was performed to evaluate whether there were statistical differences in the seroconversion rate and level distribution of RV-IgA between the two groups. Results: The seroconversion rate of serum RV-IgA in month 2 was 73.84% in the interval vaccination and 63.95% in the simultaneous vaccination group, and the difference between them was statistically significant (P<0.05). The geometric mean concentrations (GMC) of RV-IgA were 97 EU/ml and 90 EU/ml, respectively (P>0.05). Compared with the simultaneous vaccination group, the seroconversion rate and GMC of serum RV-IgA in month 12 were higher in the interval vaccination group, and the differences were statistically significant (P<0.05). Conclusions Simultaneous vaccination with live attenuated polio vaccine would affect the immune response of live attenuated rotavirus vaccine, especially the maintenance of RV-IgA antibody level.

Objective:To screen the neutralizing epitope of enterovirus 71 (EV71) and determine the specific minimum amino acid sequence that triggers immunity for providing a theoretical basis for the development of synthetic peptide vaccines.Methods:EV71 neutralizing antibody-specific binding clones were panned and sequenced using a phage display random 12-peptide library to obtain the key sequences of neutralizing epitopes. A series of peptides containing the key sequences with N-terminal acetylation (AC) and C-terminal linking to Keyhole limpet hemocyanin (KLH) were synthesized. Serum samples were collected after immunizing mice with the modified peptides. Then the immunogenicity of the peptides and the neutralizing activity of serum samples were analyzed by Western blot, ELISA and neutralization test.Results:After three rounds of panning, cloning and sequencing, KQEKDL was identified as the key motif. The serum samples collected from the mice immunized with the modified series of peptides containing key motifs had different degrees of binding ability to EV71 and VP1 protein. The serum samples of mice immunized the synthetic peptide containing only the minimum key motif (AC-KQEKDL-KLH) had the strongest response to the other three peptides and EV71 and the highest neutralizing titer.Conclusions:The EV71 neutralizing epitope was successfully screened using the phage display random peptide library. The key motif of KQEKDL might be the specific minimum amino acid sequence that triggered the immune system. This study provides a theoretical basis for better understanding the immune response mechanism, evaluating the immunogenicity of the antigens and further research and development of polypeptide vaccines.

This paper reports a case of a male patient with diabetes for more than 20 years who came to see doctor due to weakness, poor appetite and significantly elevated blood glucose. At first, it was considered that the poor blood glucose control of diabetes was the reason, and the possibility of diabetes related complications already existed. Hospital routine examination indicated a globulin increase and immunologic examination indicated that IgM was mainly increased. Subsequent imageological diagnosis revealed a small amount of bilateral pleural effusion, and after 2 weeks of hypoglycemic treatment, the blood glucose was significantly improved, but the symptoms still existed. In combination with the patient′s laboratory examination, considering the combination of blood system diseases, the hematology department was invited for consultation. Immunoelectrophoresis, bone marrow cytology, and flow cytometry were further conducted, and the patient was diagnosed with Waldenstrom macroglobulinebia. Bortezomib + dexamethasone regimen was given for chemotherapy. Currently, the patient has received 1 course of chemotherapy, and the symptoms of poor appetite were significantly improved. Coexistence of diabetes mellitus and Waldenstrom macroglobulinebia is very rare in clinical practice, and it is easy to be misdiagnosed as a complication of diabetes mellitus, both of which can be manifested as poor appetite, weakness, peripheral neuropathy and other symptoms. When diabetic patients have abnormal immunoglobulin increases and other abnormal test results that cannot be well explained by diabetes, other potential disorders, such as hematological diseases, should be considered to avoid missed diagnosis.

Objective:To analyze the relationship between plasma fibrinogen(FIB) within normal range and microalbuminuria in elderly patients with type 2 diabetes mellitus.Methods:A total of 869 elderly subjects with type 2 diabetes mellitus admitted to the Department of Endocrinology of Shanghai Fifth People′s Hospital from October 2012 to October 2014 were included in the study. The patients were divided into four groups based on the quartile level of FIB: Q1 group(<2.42 g/L), Q2 group(2.42-2.89 g/L), Q3 group(2.90-3.61 g/L), and Q4 group(≥3.62 g/L). The relationship between FIB and urinary albumin/creatinine ratio(UACR) was analyzed.Results:With the increasing of FIB, the level of UACR was significantly elevated( P<0.05). Pearson correlation analysis showed that FIB was positively associated with age, duration of diabetes, creatinine(Cr) and UACR in men and women( P<0.01). Multiple regression analysis showed that FIB was an independent factor of UACR( P<0.01). Logistic regression analysis showed that the risks of microalbuminuria and macroalbuminuria were respectively 4.536 folds(95% CI 2.516-8.175, P<0.01) and 13.314 folds(95% CI 2.925-60.612, P<0.01) in Q4 group, and 2.177 folds(95% CI 1.273-3.724, P<0.01) and 4.098 folds(95% CI 1.101-19.226, P<0.05) in Q3 group as compared with Q1 group after adjused by following factors: gender, age, duration of diabetes, body mass index(BMI), systolic blood pressure(SBP), diastolic blood pressure(DBP), fasting plasma glucose(FPG), HbA 1C, total cholesterol(TC), triglyceride(TG), low density lipoprotein-cholesterol(LDL-C), Cr, alanine aminotransferase(ALT), as well as smoking and drinking behavior. Based on the cut off values to UACR 30 mg/g and 300 mg/g, the receiver operating characteristic curve(ROC) was used to evaluate the value of FIB for UACR. The optimal cut-off value of FIB was 3.18 g/L and 3.22 g/L respectively. Conclusions:Plasma FIB was closely associated with microalbuminuria in elderly patients with type 2 diabetes mellitus, which may be considered as one of the predictors for diabetic nephropathy.

OBJECTIVE: To explore the clinical features, lysosomal enzymatic [acid α-glucosidase (GAA)] activities and genetic variants in a child with late-onset Pompe disease (LOPD). METHODS: Clinical data of a child who had presented at the Genetic Counseling Clinic of West China Second University Hospital in August 2020 was retrospectively analyzed. Blood samples were collected from the patient and her parents for the isolation of leukocytes and lymphocytes as well as DNA extraction. The activity of lysosomal enzyme GAA in leukocytes and lymphocytes was analyzed with or without addition of inhibitor of GAA isozyme. Potential variants in genes associated with neuromuscular disorders were analyzed, in addition with conservation of the variant sites and protein structure. The remaining samples from 20 individuals undergoing peripheral blood lymphocyte chromosomal karyotyping were mixed and used as the normal reference for the enzymatic activities. RESULTS: The child, a 9-year-old female, had featured delayed language and motor development from 2 years and 11 months. Physical examination revealed unstable walking, difficulty in going upstairs and obvious scoliosis. Her serum creatine kinase was significantly increased, along with abnormal electromyography, whilst no abnormality was found by cardiac ultrasound. Genetic testing revealed that she has harbored compound heterozygous variants of the GAA gene, namely c.1996dupG (p.A666Gfs*71) (maternal) and c.701C>T (p.T234M) (paternal). Based on the guidelines from the American College of Medical Genetics and Genomics, the c.1996dupG (p.A666Gfs*71) was rated as pathogenic (PVS1+PM2_Supporting+PM3), whilst the c.701C>T (p.T234M) was rated as likely pathogenic (PM1+PM2_Supporting+PM3+PM5+PP3). The GAA in the leukocytes from the patient, her father and mother were respectively 76.1%, 91.3% and 95.6% of the normal value without the inhibitor, and 70.8%, 112.9% and 128.2% of the normal value with the inhibitor, whilst the activity of GAA in their leukocytes had decreased by 6 ~ 9 times after adding the inhibitor. GAA in lymphocytes of the patient, her father and mother were 68.3%, 59.0% and 59.5% of the normal value without the inhibitor, and 41.0%, 89.5% and 57.7% of the normal value with the inhibitor, the activity of GAA in lymphocytes has decreased by 2 ~ 5 times after adding the inhibitor. CONCLUSION The child was diagnosed with LOPD due to the c.1996dupG and c.701C>T compound heterozygous variants of the GAA gene. The residual activity of GAA among LOPD patients can range widely and the changes may be atypical. The diagnosis of LOPD should not be based solely on the results of enzymatic activity but combined clinical manifestation, genetic testing and measurement of enzymatic activity.
Humans ; Child ; Male ; Female ; Glycogen Storage Disease Type II/pathology* ; Retrospective Studies ; alpha-Glucosidases/genetics* ; Mothers ; Lysosomes/pathology* ; Mutation

Objective To explore the relationship between hemoglobin (Hb) level during the first trimester of pregnancy and gestational diabetes mellitus (GDM). Methods A total of 1 276 participants, who underwent scheduled prenatal examination and normal singleton delivery at the Fifth People′s Hospital of Shanghai and Hospital of Intergrated Chinese and Western Medicine in Minhang District, from January 2016 to May 2018 were included. There were 99 cases of GDM (GDM group) and 1 177 cases of normal (control group) pregnant women.Based on the serum Hb level during the first trimester of pregnancy, participants were divided into three groups, 236 cases of low Hb level group (Hb<110 g/L), 868 cases of normal Hb level group (110 g/L≤Hb<130 g/L), and 172 cases of high Hb level group (Hb≥130 g/L). Maternal clinical data were collected, including Hb level during the first trimester of pregnancy, three-point blood glucose (BG) of oral glucose tolerance test (OGTT) and fasting insulin during the second trimester of pregnancy. Homeostasis model assessment of insulin resistance index (HOMA-IR) and homeostasis model assessment of pancreatic β cell function index (HOMA-β) were used to evaluate insulin resistance and pancreatic β cell function. Results (1) Hb level during the first trimester of pregnancy in GDM group was significantly higher than that in control group [(123±10),(119±11) g/L, P<0.05]. There were no significant difference in gravidity, parity, index of liver and renal function (all P>0.05). (2) Pre-pregnancy body mass index (BMI), 1-hour BG and 2-hour BG of OGTT were significantly increased in the high Hb level group during the first trimester of pregnancy, which were (23±4) kg/m2, (7.3±2.0) mmol/L, and (6.5±1.4) mmol/L (P<0.05), respectively. The pre-pregnancy BMI, 1-hour BG and 2-hour BG of the normal or low Hb level group were (22±3) kg/m2, (6.7±1.6) mmol/L, (6.1± 1.2) mmol/L; (22±3) kg/m2, (6.5±1.5) mmol/L, (5.9±1.1) mmol/L, respectively. There were no statistically significant difference in levels of fasting blood glucose, fasting insulin, HOMA-IR and HOMA-β within 3 groups (all P>0.05). (3) In the high Hb level group, prevalence of pregnancy overweight or obesity and GDM were the highest, which were 37.2%(64/172) and 15.1%(26/172), respectively; the differences were statistically significant (all P<0.05). (4) The serum Hb level in the first trimester was positively related with pre-pregnancy BMI (r=0.130, P<0.05), 1-hour BG (r=0.129, P<0.05), 2-hour BG (r=0.134, P<0.05), fasting insulin (r=0.096, P<0.05), and HOMA-IR (r=0.101, P<0.05).Logistic regression indicated that Hb≥130 g/L during the first trimester of pregnancy was an independent risk factor for GDM ( OR=2.799, 95% CI :1.186-6.604; P<0.05). Conclusion The high level of Hb (Hb≥130 g/L) during the first trimester of pregnancy is associated with GDM.