This study aims to explore the effects and action mechanisms of the active ingredients in Buyang Huanwu Decoction(BYHWD), namely tetramethylpyrazine(TMP) and hydroxy-safflor yellow A(HSYA), on oxygen-glucose deprivation/reglucose-reoxygenation(OGD/R)-induced inflammation and oxidative stress of microglia(MG). Network pharmacology was used to screen the effective monomer ingredients of BYHWD and determine the safe concentration range for each component. Inflammation and oxidative stress models were established to further screen the best ingredient combination and optimal concentration ratio with the most effective anti-inflammatory and antioxidant effects. OGD/R BV2 cell models were constructed, and BV2 cells in the logarithmic growth phase were divided into a normal group, a model group, an HSYA group, a TMP group, and an HSYA + TMP group. Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of inflammatory cytokines such as interleukin-1β(IL-1β), tumor necrosis factor-α(TNF-α), and interleukin-6(IL-6). Oxidative stress markers, including superoxide dismutase(SOD), nitric oxide(NO), and malondialdehyde(MDA), were also measured. Western blot was used to analyze the protein expression of both inflammation-related pathway [Toll-like receptor 4(TLR4)/nuclear factor-kappa B(NF-κB)] and oxidative stress-related pathway [nuclear factor erythroid 2-related factor 2(Nrf2)/heme oxygenase-1(HO-1)]. Immunofluorescence was used to assess the expression of proteins such as inducible nitric oxide synthase(iNOS) and arginase-1(Arg-1). The most effective ingredients for anti-inflammatory and antioxidant effects in BYHWD were TMP and HSYA. Compared to the normal group, the model group showed significantly increased levels of IL-1β, TNF-α, IL-6, NO, and MDA, along with significantly higher protein expression of NF-κB, TLR4, Nrf2, and HO-1 and significantly lower SOD levels. The differences between the two groups were statistically significant. Compared to the model group, both the HSYA group and the TMP group showed significantly reduced levels of IL-1β, TNF-α, IL-6, NO, and MDA, lower expression of NF-κB and TLR4 proteins, higher levels of SOD, and significantly increased protein expression of Nrf2 and HO-1. Additionally, the expression of the M1-type MG marker iNOS was significantly reduced, while the expression of the M2-type MG marker Arg-1 was significantly increased. The results of the HSYA group and the TMP group had statistically significant differences from those of the model group. Compared to the HSYA group and the TMP group, the HSYA + TMP group showed further significant reductions in IL-1β, TNF-α, IL-6, NO, and MDA levels, along with significant reductions in NF-κB and TLR4 protein expression, an increase in SOD levels, and elevated Nrf2 and HO-1 protein expression. Additionally, the expression of the M1-type MG marker iNOS was reduced, while the M2-type MG marker Arg-1 expression increased significantly in the HSYA + TMP group compared to the TMP or HSYA group. The differences in the results were statistically significant between the HSYA + TMP group and the TMP or HSYA group. The findings indicated that the combined use of HSYA and TMP, the active ingredients of BYHWD, can effectively inhibit OGD/R-induced inflammation and oxidative stress of MG, showing superior effects compared to the individual use of either component.
Oxidative Stress/drug effects* ; Drugs, Chinese Herbal/pharmacology* ; Animals ; Mice ; Glucose/metabolism* ; Cell Line ; Inflammation/genetics* ; Oxygen/metabolism* ; Pyrazines/pharmacology* ; Microglia/metabolism* ; NF-E2-Related Factor 2/immunology* ; NF-kappa B/immunology* ; Toll-Like Receptor 4/immunology* ; Anti-Inflammatory Agents/pharmacology* ; Humans

Nonobstructive azoospermia (NOA), one of the most severe types of male infertility, etiology often remains unclear in most cases. Therefore, this study aimed to detect four biallelic detrimental variants (0.5%) in the minichromosome maintenance domain containing 2 ( MCMDC2 ) genes in 768 NOA patients by whole-exome sequencing (WES). Hematoxylin and eosin (H&E) demonstrated that MCMDC2 deleterious variants caused meiotic arrest in three patients (c.1360G>T, c.1956G>T, and c.685C>T) and hypospermatogenesis in one patient (c.94G>T), as further confirmed through immunofluorescence (IF) staining. The single-cell RNA sequencing data indicated that MCMDC2 was substantially expressed during spermatogenesis. The variants were confirmed as deleterious and responsible for patient infertility through bioinformatics and in vitro experimental analyses. The results revealed four MCMDC2 variants related to NOA, which contributes to the current perception of the function of MCMDC2 in male fertility and presents new perspectives on the genetic etiology of NOA.
Humans ; Male ; Azoospermia/genetics* ; Meiosis/genetics* ; Spermatogenesis/genetics* ; Adult ; Exome Sequencing ; Microtubule-Associated Proteins/genetics* ; Alleles ; Infertility, Male/genetics*

OBJECTIVE: To examine the anti-inflammatory effect of grape seed extract (GSE) in animal and cellular models and explore its mechanism of action. METHODS: This study determined the inhibitory effect of GSE on macrophage inflammation and Th1 and Th17 polarization in vitro. Based on the in vitro results, the effects and mechanisms of GSE on multiple sclerosis (MS)-experimental autoimmune encephalomyelitis (EAE) mice model were further explored. The C57BL/6 mice were intragastrically administered with 50 mg/kg of GSE once a day from the 3rd day to the 27th day after immunization. The activation of microglia, the polarization of Th1 and Th17 and the inflammatory factors such as tumor necrosis factor- α (TNF- α), interleukin-1 β (IL-1 β), IL-6, IL-12, IL-17 and interferon-γ (IFN-γ) secreted by them were detected in vitro and in vivo by flow cytometry, enzyme linked immunosorbent assay (ELISA), immunofluorescence staining and Western blot, respectively. RESULTS: GSE reduced the secretion of TNF-α, IL-1 β and IL-6 in bone marrow-derived macrophages stimulated by lipopolysaccharide (P<0.01), inhibited the secretion of TNF-α, IL-1 β, IL-6, IL-12, IL-17 and IFN-γ in spleen cells of EAE mice immunized for 9 days (P<0.05 or P<0.01), and reduced the differentiation of Th1 and Th17 mediated by CD3 and CD28 factors (P<0.01). GSE significantly improved the clinical symptoms of EAE mice, and inhibited spinal cord demyelination and inflammatory cell infiltration. Peripherally, GSE downregulated the expression of toll-like-receptor 4 (TLR4) and Rho-associated kinase (ROCKII, P<0.05 or P<0.01), and inhibited the secretion of inflammatory factors (P<0.01 or P<0.05). In the central nervous system, GSE inhibited the infiltration of CD45⁺CD11b⁺ and CD45⁺CD4⁺ cells, and weakened the differentiation of Th1 and Th17 (P<0.05). Moreover, it reduced the secretion of inflammatory factors (P<0.01), and prevented the activation of microglia (P<0.05). CONCLUSION GSE had a beneficial effect on the pathogenesis and progression of EAE by inhibiting inflammatory response as a potential drug and strategy for the treatment of MS.
Mice ; Animals ; Encephalomyelitis, Autoimmune, Experimental/pathology* ; Grape Seed Extract/therapeutic use* ; Interleukin-17 ; Interleukin-1beta ; Tumor Necrosis Factor-alpha/metabolism* ; Interleukin-6/metabolism* ; Th1 Cells ; Mice, Inbred C57BL ; Interferon-gamma/therapeutic use* ; Th17 Cells/metabolism* ; Interleukin-12/therapeutic use* ; Cytokines/metabolism*

Objective - To analyze the prevalence and influencing factors of work related musculoskeletal disorders (WMSDs) Methods among painters in the manufacturing industry. A total of 639 painters from one shipbuilding enterprise, one automobile manufacturing enterprise and three wooden furniture manufacturing enterprises in Guangdong Province were selected as the research subjects using typical sampling method. The Chinese version of Musculoskeletal Disorders Questionnaire was Results used to investigate the prevalence of WMSDs in the past one year, and the influencing factors were analyzed. The total prevalence rate of WMSDs among painters in the manufacturing industry was 37.4%. The prevalence of WMSDs in different vs vs P industries from high to low was shipbuilding, automobile and furniture manufacturing (50.0% 38.7% 29.0%, <0.01). The prevalence of WMSDs in different parts of the body from high to low was neck, ankle/foot, shoulder, low back, upper back, knee, vs vs vs vs vs vs vs vs P hand/wrist, hip/leg and elbow (20.7% 19.2% 17.4% 15.8% 14.1% 13.8% 13.5% 9.5% 6.6%, <0.01). Multivariate logistic regression analysis results showed that working in uncomfortable postures was a risk factor for neck, ankle/ P P foot and shoulder WMSDs (all <0.01); long time head turning was a risk factor for neck and shoulder WMSDs (both <0.05); P overweight and obesity, and bending and turning frequently at the same time were risk factors for ankle/foot WMSDs (all <0.05); P adequate rest time was a protective factor for neck and ankle/foot WMSDs (both <0.01); participated in physical exercise more P than once a week was a protective factor of neck and shoulder WMSDs in painters (all <0.05), after excluding the influence of Conclusion confounding factors. The prevalence of WMSDs in manufacturing painters was high, and the main body parts E mail 4813545@qq.com E mail wangzhongxu2003@163.com· · 中国职业医学 年 月第 卷第 期 ， ， ， 482 2022 10 49 5 Chin Occup Med October 2022 Vol.49 No.5 involved were neck, ankle/foot and shoulder. The influencing factors include individual factors, poor ergonomics factors and unreasonable work organization.

Objective To identify new genes that correlate with prognosis of clear-cell renal cell carcinoma (ccRCC)

Objective: To investigate the safety and efficacy of regional transport to percutaneous coronary intervention(PCI) hospitals from non-PCI hospitals after thrombolysis in patients with acute ST-segment elevation myocardial infarction(STEMI) in northwest China. Methods: In this retrospective study, 1 062 STEMI patients who were transferred from non-PCI hospitals within 24 hours from symptom onset, during January 2015 and January 2019 in the First Hospital of Lanzhou University, were included. According to the treatment strategy, they were divided into two groups, namely intravenous thrombolysis combined with PCI group(n=240), and primary PCI group(n=822). Observation endpoint were in-hospital adverse cardiovascular and cerebrovascular events and bleeding events, Including all-cause death, ischemic stroke, malignant arrhythmia, intracranial hemorrhage and hemorrhage with hemoglobin decrease≥50 g/L. Results: A total of 1 062 STEMI patients were included(age was (61±12) years old), with 905 males (85.2%). The proportion of grade 0 TIMI blood flow in the primary PCI group before operation was significantly higher than that in the thrombolysis combined with PCI group(63.0%(518/822) vs. 36.3%(87/240)， P<0.001). Compared with primary PCI group, the time from symptom onset to first medical contact(2.11(1.00, 4.00)hours vs.3.00(1.13, 7.07)hours, P<0.001) and reperfusion in thrombolysis combined with PCI group(3.07(1.83, 4.87)hours vs. 6.92(4.07, 11.15) hours, P<0.001) were significantly shorter. The proportion of all-cause death was significantly higher in the primary PCI group than that in the thrombolysis combined with PCI group (1.8%(15/822) vs. 0, P=0.03). There was no significant difference in hemorrhage, ischemic stroke and malignant arrhythmia between the two groups(all P>0.05). Conclusions: For STEMI patients initially hospitalized in non-PCI hospitals, regional transport combined with PCI is feasible and effective. It does not significantly increase the risk of bleeding and cardiovascular and cerebrovascular events, with shorter time from symptom onset to myocardial reperfusion.
Aged ; Angioplasty, Balloon, Coronary ; China ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; Percutaneous Coronary Intervention ; Retrospective Studies ; ST Elevation Myocardial Infarction/therapy* ; Thrombolytic Therapy ; Treatment Outcome

Kidney stones are a common urinary system condition that can progress to kidney disease. Previous studies on the association between tea consumption and kidney stones are inconsistent. A cross-sectional study to investigate the association between tea consumption and kidney stones was conducted from 2013 to 2014 and recruited 9,078 northern Chinese adults. A total of 8,807 participants were included in the final analysis. Participants' prevalence of kidney stones was 1.07%, 1.73%, and 2.25% based on their tea consumption frequency of never, occasionally, and often groups, respectively. Compared with the 'never' group, the odds ratios (95% confidence intervals) for the occurrence of kidney stones were 1.57 (1.00-2.46) and 1.65 (1.06-2.57) in the 'occasionally' and 'often' groups, respectively. After adjusting for sex, age, and other potential confounding factors, tea consumption still significantly increased the risk of kidney stones. Tea consumption is independently associated with an increased risk of kidney stones in the investigated population, suggesting that a decrease in the consumption of tea may be a preventive strategy for kidney stones.

BACKGROUND: Studies have shown that bone marrow mesenchymal stem cells have the potential of differentiation into alveolar epithelial cells in vitro, but so far no study has indicated that adipose-derived mesenchymal stem cells (ADSCs) can be differentiated into alveolar epithelial cells through long-term Transwell co-culture. OBJECTIVE: To observe whether rat lung epithelial-T-antigen negative cell lines (RLE-6TN) can induce rat ADSCs to differentiate into type II alveolar epithelial cells by long-term Transwell co-culture. METHODS: Three SPF health female Sprague-Dawley rats were used as donors to separate, extract, culture and identity ADSCs. The experimental group was subjected to the Transwell co-culture of ADSCs and RLE-6TN, while the control group was subjected to the culture of ADSCs alone. The morphological changes of ADSCs were observed by the inverted phase contrast microscope at 21 days after co-culture. Immunofluorescence staining using surfactant protein C (SP-C) was performed on the co-cultured ADSCs. The fluorescence staining was observed using the inverted fluorescence microscope. Integral optical density (IOD) analysis was conducted by Image pro plus 6.0 software. RESULTS AND CONCLUSION: RLE-6TN cells were identified by fluorescence staining with stable expression of SP-C protein (red fluorescence) in the experimental group, and there was no red fluorescence in the control group. After 21-day co-culture, the cell shape in the experimental group was transformed from the long spindle shape into oval or polygon shape gradually, while the cell shape in the control group remained fibroblast-like. These results show that RLE-6TN can induce ADSCs to differentiate into type II alveolar epithelial cells after a long-term (21 days) co-culture.

ObjectiveTo explore the application value of real-time contrast-enhanced ultrasound (RTCEU) in improving the detection rate of transrectal ultrasound-guided prostate biopsy.

METHODSThis prospective study included 91 male patients with abnormally high PSA (4－20 μg/L) or abnormalities in DRE or MRI, who underwent 12+X prostate biopsy following conventional transrectal ultrasonography (TRUS) and RTCEU examination. We compared the numbers of suspected prostatic nodules before and after RTCEU as well as the detection rates of prostate cancer between conventional TRUS-guided 12PBx and 12PBx plus lesion-targeted biopsy procedures.

RESULTSTotally, 57 of the 86 suspected lesions on TRUS (66.3%), and 108 of the 118 abnormal nodules on RTCEU (91.5%) were confirmed to be prostate cancer. RTCEU achieved a significantly higher detection rate than TRUS (P<0.01). A total of 39 cases of prostate cancer (42.8%) were detected by RTCEU, while only 28 (30.7%) by TRUS, with statistically significant difference in the detection rate between the two procedures (P=0.033).

CONCLUSIONSReal-time contrast-enhanced ultrasound can significantly improve the detection rate of prostate cancer and provide a valuable guide to targeted prostate biopsy.

Contrast Media ; Endoscopic Ultrasound-Guided Fine Needle Aspiration ; Humans ; Magnetic Resonance Imaging ; Male ; Prospective Studies ; Prostate ; diagnostic imaging ; pathology ; Prostate-Specific Antigen ; blood ; Prostatic Neoplasms ; blood ; diagnostic imaging ; pathology ; Ultrasonography, Interventional

OBJECTIVETo investigate the clinical, pathological and immunohistochemical features of vulvar intraepithelial neoplasia (VIN).

METHODSAccording to the 2004 modified terminology of International Society for the Study of Vulvovaginal Diseases (ISSVD), the cases were diagnosed as VIN from patients who had performed vulvar biopsy in Beijing Wuzhou Women's Hospital from February 2009 to December 2011, which were reclassified as usual VIN and differentiated VIN. The clinical and pathological studies were conducted respectively. MaxVision immunohistochemical staining was used to detect the expression of Ki-67, p16 and p53 proteins.

RESULTSThere were 20 cases of VIN in 237 patients, and the incidence of VIN was 8.4% in all of contemporary vulvar biopsy. In 17 cases of usual VIN, mean age was 29.6 years, the lesion typically presented with atypical cells involving almost all layers of the epithelium, which was equivalent to the high-grade squamous intraepithelial neoplasia of cervix. Immunohistochemistry for Ki-67 and p16 was strongly positive in usual VIN. High risk human papillomavirus (HPV) detection was also positive. The incidence of differentiated VIN was less than usual VIN, and there were only 3 cases in this study. In differentiated VIN, patients aged over 50 years, with mean of 53.7 years, and the lesion most commonly presented with lichen sclerosis background. There were epithelial thickening and extending, and parakeratosis, and atypia was strictly confined to the basal and parabasal layers of the epithelium where the cells enlarged with abundant eosinophilic cytoplasm, presented with prominent nucleoli, increased cellularity and abnormal keratinization. In differentiated VIN, p53 was strongly positive, Ki-67 and p16 immunohistochemical expression was confined to the basal layer only.

CONCLUSIONSVIN is a precursor of invasive squamous cell carcinoma of the vulva. The modified terminology of ISSVD classifies VIN as high-grade lesions. Definitive pathological diagnosis of VIN plays an important role in its timely treatment and the prevention of vulvar carcinoma.

Adult ; Carcinoma in Situ ; metabolism ; pathology ; virology ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; Female ; Humans ; Immunohistochemistry ; Ki-67 Antigen ; metabolism ; Middle Aged ; Papillomavirus Infections ; pathology ; Tumor Suppressor Protein p53 ; metabolism ; Vulvar Neoplasms ; metabolism ; pathology ; virology ; Young Adult