OBJECTIVETo observe the influence of different concentrations of bisphenol A (BPA) on glucose metabolism and lactate dehydrogenase (LDH) expression in rat Sertoli cells in vitro and investigate the mechanisms of BPA inducing male infertility.

METHODSUsing two-step enzyme digestion, we isolated Sertoli cells from male Wistar rats and constructed a primary Sertoli cell system, followed by immunohistochemical FasL staining. We randomly divided the Sertoli cells into a control group to be cultured in the serum-free minimal essential medium (MEM) plus dimethyl sulfoxide (DMSO) and three experimental groups to be treated with 100 nmol/L, 10 μmol/L, and 1 mmol/L BPA, respectively, in the MEM plus DMSO. After 48 hours of treatment, we measured the proliferation of the cells by CCK-8 assay, determined the concentrations of metabolites by NMR spectroscopy, and detected the expression of LDH in the Sertoli cells by RT-PCR and Western blot.

RESULTSThe purity of the isolated Sertoli cells was (96.05 ± 1.28)% (n = 10). Compared with the control group, the 100 nmol/L, 10 μmol/L, and 1 mmol/L BPA groups showed no remarkable changes in the proliferation of Sertoli cells ([98 ± 8]%, [96 ± 3]%, and [95 ± 3]%, P >0.05), but the 10 μmol/L and 1 mmol/L of BPA groups exhibited significantly decreased concentrations of intracellular glucose ([3.89 ± 0.07] vs [3.36 ± 0.24] and [3.04 ± 0.21] pmol/cell, P <0.05) and lactate ([0.43 ± 0.06] vs [0.29 ± 0.05] and [0.20 ± 0.03] pmol/cell, P <0.05). The expression of LDH mRNA was decreased with the increased concentration of BPA, while that of LDH protein reduced only in the 1 mmol/L BPA group (P <0.05).

CONCLUSIONHigh-concentration BPA decreases the expression of LDH and alters glucose metabolism in Sertoli cells, and therefore may reduce the provision of lactate for germ cells and impair spermatogenesis.

Animals ; Benzhydryl Compounds ; administration & dosage ; pharmacology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media, Serum-Free ; Dimethyl Sulfoxide ; pharmacology ; Glucose ; metabolism ; In Vitro Techniques ; Infertility, Male ; chemically induced ; L-Lactate Dehydrogenase ; metabolism ; Male ; Phenols ; administration & dosage ; pharmacology ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Wistar ; Sertoli Cells ; drug effects ; metabolism ; Spermatogenesis ; drug effects

Objective To observe the clinical efficacy of transcatheter arterial chemoembolization (TACE) combined with radiofrequency ablation (RFA) and hepatic artery infusion of autologous cytokineinduced killer (CIK) cells in treating clinical stage I hepatocellular carcinoma (HCC).Methods A total of 80 patients with confirmed HCC,who were treated with comprehensive interventional therapy during the period from January 2009 to May 2010,were enrolled in this follow-up study.The patients were divided into the study group (n=38),receiving TACE,RFA and autologous CIK cells therapy,and the control group (n=42),receiving TACE and RFA only.The quality of life (QOL),changes in immune function indexes,progression free survival (PFS) and survival rate were calculated,and the results were compared between the two groups.Results (1) QOL score:after the treatment the QOL score of the study group was significantly higher than that of the control group (P＜0.05).(2) Immune function:the post-treatment immune function values were different from the pre-treatment ones in both groups,the differences were statistically significant (P＜0.05);and the differences between the two groups were also statistically significant (P＜0.05),with the changes of the study group being more obvious.(3) PFS and survival rate:the median PFS of the study group and the control group was 48.0 and 40.1 months respectively,while the one-,2-,3-,5-year survival rates of the study group and the control group were 100％,89.5％,71.1％,55.3％ and 95.2％,88.1％,64.3％,28.6％ respectively.Both the median PFS and survival rate in the study group were higher than those in the control group.Conclusion In treating clinical stage I HCC,TACE combined with RFA and hepatic artery infusion of autologous CIK ceils can improve QOL of patients,strengthen patient's immune function,prolong the median PFS,and increase the overall survival rate.

OBJECTIVETo evaluate the expression and clinical significance of urinary nuclear matrix protein (NMP22) and cytokeratin 18 (CK18) for transitional cell carcinoma of the bladder.

METHODSUrinary NMP22 and CK18 levels of 293 patients with transitional cell carcinoma of the bladder, 400 patients with non-transitional cell carcinoma of the bladder, and 105 bladder benign disease were analysed by enzyme-linked-immunosorbent assay (ELISA).

RESULTSThe levels of urinary NMP22 and CK18 in the patients with transitional cell carcinoma of the bladder (M = 17.3 U/ml, M(CK18) = 484.2 U/L) were significantly higher than those in the non-transitional cell carcinoma of the bladder (M = 6.8 U/ml, M(CK18) = 156.0 U/L) and the benign disease group (M(NMP22) = 2.3 U/ml, M(CK18) = 66.6 U/L) (P < 0.001). The sensitivity and specificity of urinary NMP22 and CK18 were 79.2%, 88.6% and 78.2%, 82.9%, respectively, for transitional cell carcinoma of the bladder before any treatment. The joint sensitivity of the two markers was 91.7%. The NMP22 and CK18 levels were significantly lower in the recovered patients after surgical operation (P < 0.01), while in patients with recurrence or metastasis the levels of the markers were significantly higher (P < 0.01). There was a significant relationship between NMP22 and CK18, (r = 0.689, P < 0.01). The levels of urinary nmp22 and CK18 were significantly different among pathological grade G1, G2, G3, and stage Ta, T1, T2, T3 (P < 0.01).

CONCLUSIONNMP22 and CK18 are useful tumor marker for diagnosis of transitional cell carcinoma of the bladder and for monitoring the state of illness. The joint use of the two markers can improve the sensitivity of cancer detection. NMP22 and CK18 may become a new class of tumor markers, and to be the basis for development of a new assay with an increased efficacy for the detection and treatment of bladder cancer.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; urine ; Carcinoma, Renal Cell ; urine ; Carcinoma, Transitional Cell ; diagnosis ; pathology ; surgery ; urine ; Child ; Female ; Follow-Up Studies ; Humans ; Keratin-18 ; urine ; Male ; Middle Aged ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; urine ; Neoplasm Staging ; Nuclear Proteins ; urine ; Prognosis ; Sensitivity and Specificity ; Urinary Bladder Neoplasms ; diagnosis ; pathology ; surgery ; urine ; Young Adult

Objective To investigate the Gram-positive coccus resistance in nationwide's tertiary hospitals and understand the trend of antimicrobial resistance.Methods All the clinical isolates were collected from 19 hospitals and the minimal inhibitory concentrations(MICs)were tested using agar/broth dilution method recommended.Results A total of 1 974 pathogenic Gram-positive coccus from 19 tertiary hospitals in 19 cities nationwide over the period from July 2021 to June 2022 were studied.Based on the MIC results,the prevalence of methicillin resistant Stapylococcus aureus(MRSA)and methicillin resistant Stapylococcus epidermidis(MRSE)were 36.4％and 79.9％respectively.No vancomycin insensitivity Staphylococcus was detected.Staphylococcus aureus were 100％susceptibility to linezolid and teicoplanin.Antibiotic resistance rate of Enterococcus faecalis and Enterococcus faecium to ampicillin were 3.1％and 92.9％.The detectation rate of vancomycin resistant Enterococcus(VRE)was 1.6％.Nonsusceptibility rate of Enterococcus faecalis to linezolid was 32.2％,two consecutive monitoring rises and nonsusceptibility rate of Enterococcus faecium(12.5％)was also significantly increased.The prevalence of penicillin non-susceptible Streptococcus pneumoniae(PNSSP)was 0.8％based on non-meningitis and parenteral administration criterion,decrease of nearly 30 percentage points from the previous surveillance.While for cases of oral penicillin,the rate was 71.8％,showing similar to last time.The results indicated that the number of strains with higher MIC value of penicillin(MIC ≥4 mg·L-1)decreased significantly.There were no significant differences of resistance rates of Stapylococcus aureus,Stapylococcus epidermidis,Enterococcus faecalis,Enterococcus faecium and Streptococcus pneumoniae among various groups such as different department,age,or specimen source.Conclusion VRE detection ratio stablized at a relatively low level.The number of Streptococcus pneumoniae with higher MIC value of penicillin decreased significantly compared with the previous monitoring.The increase of linezolidin-insensitive Enterococcus was noteworthy.

Objective To investigate the Gram-negative bacteria resistance in nationwide's tertiary hospitals and understand the trend of antimicrobial resistance.Method All the clinical isolates were collected from 19 hospitals and the minimal inhibitory concentrations(MICs)were tested using agar/broth dilution method recommended.Results A total of 4 066 pathogenic isolates from 19 tertiary hospitals in 19 cities nationwide over the period from July 2021 to June 2022 were studied.Based on the MIC results,Escherichia coli and Klebsiella pneumoniae showed extended spectrum β-lactamase(ESBLs)phenotype rates of 55.0％and 21.0％,respectively,ESBLs phenotype rate of Klebsiella pneumoniae keep going down.The ratios of carbapenems resistance Klebsiella pneumoniae increased by 5 percentage points compared with the previous monitoring.Carbapenems,moxalactam,sitafloxacin,β-lactam combination agents,fosfomycin trometamol,and amikacin displayed desirable antibacterial activity against Enterbacterales,susceptibal rates were above 75％.In addition,tigacycline,omacycline,colistin and fluoxefin maintained good antibacterial activity against their respective effective bacteria/species,and the bacterial sensitivity rates by more than 80％.Resistance rates of Pseudomonas aeruginosa and Acinetobacter baumannnii to imipenem were 26.3％and 72.1％and multidrug-resistant(MDR)detection rates were 41.1％and 77.3％,extensively drug-resistant(XDR)were 12.0％and 71.8％,respectively.Comparison of drug resistance rates from different wards,ages and specimen sources indicated that the proportion of resistance in Klebsiella pneumoniae and Acinetobacter baumannii isolated from intensive care unit(ICU)were significantly higher than non-ICU.Carbapenem resistance rates of Klebsiella pneumoniae isolated from ICU were more than 35％.Resistance rates of Haemophilus influenzae isolated in children to β-lactam,macrolide,clindamycin and ESBLs detection rate in Klebsiella pneumoniae isolated from children were more than those from adults and the old people,so bacterial resistance in children is an important problem in China.Conclusion ESBLs detection rate of Escherichia coli increased slightly after years of continuous decline.The proportion of carbapenem resistant Pseudomonas aeruginosa was stable,but the resistance rate of Klebsiella pneumoniae and Acinetobacter baumannii to carbapenems was still increased,which should be paid more attention.

OBJECTIVETo explore the mechanism of hyperthermia inducing infertility by observing the expression of glial cell line-derived neurotrophic factor (GDNF) in rat Sertoli cells cultured in vitro at different temperatures.

METHODSUsing combination enzyme digestion and selective adhesion, we isolated Sertoli cells from male Wistar rats and cultured them in vitro at different temperatures, followed by observation of the changes in their adhesion and morphology and identification by FasL immunohistochemical staining. We divided the Sertoli cells into a control group (35 degrees C) and four experimental groups (36 degrees C, 37 degrees C, 38 degrees C, and 39 degrees C), measured their proliferation by CCK-8, observed their morphology and structure by HE staining, and determined the expression of GDNF by RT-PCR, immunofluorescence and Western blot.

RESULTSSertoli cells were successfully isolated and in vitro-cultured, with a purity of (95.30 +/- 2.15)% (n = 10). The CCK-8 assay showed that the proliferation of the Sertoli cells was the highest at 36 degrees C, gradually decreasing with the temperature above 36 degrees C, and significantly inhibited at 39 degrees C (P < 0.01). Immunofluorescence revealed the expression of GDNF in the cytoplasm, with the highest fluorescence intensity at 36 degrees C. RT-PCR and Western blot exhibited a decreasing trend of the GDNF expression with the increasing temperature above 36 degrees C. There were statistically significant differences in the expression of GDNF between the control group and the four experimental groups (P < 0.01).

CONCLUSIONThe proliferation and GDNF expression of in vitro-cultured Sertoli cells differ significantly at different temperatures. At > 36 degrees C, the higher the temperature is, the lower the Sertoli cell proliferation and GDNF expression are. Our findings suggest that high temperature above 36 degrees C suppresses the function of Sertoli cells and may also damage spermatogenesis.

Animals ; Cells, Cultured ; Glial Cell Line-Derived Neurotrophic Factor ; metabolism ; Male ; Rats ; Rats, Wistar ; Sertoli Cells ; cytology ; metabolism ; Temperature ; Testis ; cytology

OBJECTIVE: To analyze the main factors affecting the positive results of acupuncture for chronic pain in English literature of randomized controlled trial (RCT), in order to provide reference for the design of acupuncture clinical research. METHODS: The RCTs of acupuncture for chronic pain published before March 26, 2020 were searched in PubMed, EMbase and Cochrane Library by computer. A total of 21 factors were analyzed by single-factor analysis, and the factors with statistically significant difference were selected for multivariate Logistic regression analysis. RESULTS: A total of 69 RCTs were included, including 47 RCTs (68.12%) with positive results and 22 RCTs (31.88%) with non-positive results. The multivariate Logistic regression analysis was performed with the three screened factors (publication year, treatment frequency and intervention form) selected by single-factor analysis, and the results showed that the positive results were related to the frequency of acupuncture treatment. The positive rate of RCT with frequency≥2 times a week was 3.24 times of that with frequency<2 times a week (OR=3.24, 95%CI =[1.07,9.83], P<0.05). CONCLUSION Acupuncture frequency may be the main factor affecting the positive results of RCT in English literature of acupuncture for chronic pain. More researches are needed in the future to explore the influence of acupuncture frequency on the curative effect.
Acupuncture ; Acupuncture Therapy ; Chronic Pain/therapy* ; Humans ; PubMed