Comprehensive assessment of mismatch repair and microsatellite instability status in molecular classification of endometrial carcinoma / 中华妇产科杂志

Objective: To explore the concordance and causes of different mismatch repair (MMR) and microsatellite instability (MSI) detection results in endometrial carcinoma (EC) molecular typing. Methods: A total of 214 EC patients diagnosed from January 2021 to April 2023 were selected at the Department of Pathology, Peking University Third Hospital. The immunohistochemistry (IHC) results of MMR protein were reviewed. Tumor specific somatic mutations, MMR germline mutations, microsatellite scores and tumor mutation burden (TMB) were detected by next-generation sequencing (NGS) with multi-gene panel. Methylation-specific PCR was used to detect the methylation status of MLH1 gene promoter in cases with deficient MLH1 protein expression. In cases with discrepant results between MMR-IHC and MSI-NGS, the MSI status was detected again by PCR (MSI-PCR), and the molecular typing was determined by combining the results of TMB and MLH1 gene promoter methylation. Results: (1) In this study, there were 22 cases of POLE gene mutation subtype, 55 cases of mismatch repair deficient (MMR-d) subtype, 29 cases of p53 abnormal subtype, and 108 cases of no specific molecular profile (NSMP). The median age at diagnosis of MMR-d subtype (54 years old) and the proportion of aggressive histological types (40.0%, 22/55) were higher than those of NSMP subtype [50 years old and 12.0% (13/108) respectively; all P<0.05]. (2) Among 214 patients, MMR-IHC test showed that 153 patients were mismatch repair proficient (MMR-p), 49 patients were MMR-d, and 12 patients were difficult to evaluate directly. MSI-NGS showed that 164 patients were microsatellite stable (MSS; equal to MMR-p), 48 patients were high microsatellite instability (MSI-H; equal to MMR-d), and 2 patients had no MSI-NGS results because the effective sequencing depth did not meet the quality control. The overall concordance between MMR-IHC and MSI-NGS was 94.3% (200/212). All the 12 discrepant cases were MMR-d or subclonal loss of MMR protein by IHC, but MSS by NGS. Among them, 10 cases were loss or subclonal loss of MLH1 and (or) PMS2 protein. Three discrepant cases were classified as POLE gene mutation subtype. In the remaining 9 cases, 5 cases and 3 cases were confirmed as MSI-H and low microsatellite instability (MSI-L) respectively by MSI-PCR, 6 cases were detected as MLH1 gene promoter methylation and 7 cases demonstrated high TMB (>10 mutations/Mb). These 9 cases were classified as MMR-d EC. (3) Lynch syndrome was diagnosed in 27.3% (15/55) of all 55 MMR-d EC cases, and the TMB of EC with MSH2 and (or) MSH6 protein loss or associated with Lynch syndrome [(71.0±26.2) and (71.5±20.1) mutations/Mb respectively] were significantly higher than those of EC with MLH1 and (or) PMS2 loss or sporadic MMR-d EC [(38.2±19.1) and (41.9±24.3) mutations/Mb respectively, all P<0.01]. The top 10 most frequently mutated genes in MMR-d EC were PTEN (85.5%, 47/55), ARID1A (80.0%, 44/55), PIK3CA (69.1%, 38/55), KMT2B (60.0%, 33/55), CTCF (45.5%, 25/55), RNF43 (40.0%, 22/55), KRAS (36.4%, 20/55), CREBBP (34.5%, 19/55), LRP1B (32.7%, 18/55) and BRCA2 (32.7%, 18/55). Concurrent PTEN, ARID1A and PIK3CA gene mutations were found in 50.9% (28/55) of MMR-d EC patients. Conclusions: The concordance of MMR-IHC and MSI-NGS in EC is relatively high.The discordance in a few MMR-d EC are mostly found in cases with MLH1 and (or) PMS2 protein loss or MMR protein subclonal staining caused by MLH1 gene promoter hypermethylation. In order to provide accurate molecular typing for EC patients, MLH1 gene methylation, MSI-PCR, MMR gene germline mutation and TMB should be combined to comprehensively evaluate MMR and MSI status.

Effects of long-chain noncoding RNA Linc00673 overexpression on proliferation and apoptosis of gastric cancer cell line MGC-803 / 中国应用生理学杂志

Objective: To investigate the effects of long-chain noncoding RNA Linc00673 overexpression on proliferation and apoptosis of gastric cancer cells and its mechanisms. Methods: The recombinant lentivirus expressing plasmid pLVX-Linc00673 and the control empty plasmid pLVX-NC were packaged and amplified in 293T cells, and the recombinant lentivirus was transfected into gastric cancer cell line MGC-803 to establish a cell line stably overexpressing Linc00673. The expression of Linc00673 gene was detected by real-time fluorescence quantitative PCR. The growth and proliferation of cells were observed by MTT assay and clone formation assay. Cell cycle and apoptosis were detected by flow cytometry. The expressions of cell cycle related regulatory genes were detected by qPCR. The expressions of key molecules in the PI3K/Akt signaling pathway and tumor proliferation related proteins were detected by Western blot. Results: The expressions of Linc00673 in gastric cancer cell line MGC-803, BGC-823 and AGS were significantly higher than that in normal gastric mucosa cell line GES-1 (P＜0.05). MGC-803 cell line with stable overexpression of LINC00673 was established, and the expression level of LincC00673 was 200 times higher than that of the control empty carrier group. Overexpression of Linc00673 promoted proliferation of MGC-803 cells (P＜0.05) and clone formation (P＜0.05), inhibited cell apoptosis and affected the G1→S phase progression of cell cycle (P＜0.01). Overexpression of Linc00673 could affect the expressions of cell cycle regulatory gene CCNG2, P19 and CDK1 in MGC-803. Western blot showed that Linc00673 overexpression not only promoted the expressions of the key molecule pAkt in PI3K / Akt signaling pathway and its downstream target NF-κ B and Bcl-2 protein, but also up regulated the expressions of tumor related factors β-catenin and EZH2 proteins. Conclusion: Overexpression of Linc00673 may promote proliferation and inhibit apoptosis of MGC-803 cells through PI3K/Akt signaling pathway.

Overexpression of LncRNA SLC25A25-AS1 Inhibits Proliferation‚ Migration‚ and Invasion of HeLa Cells / 中国生物化学与分子生物学报

Long non-coding RNA SLC25A25-AS1 has a tumor inhibition effect in the development of colorectal cancer. However‚ the mechanism of SLC25A25-AS1 in cervical cancer needs further study. We studied the abnormal expression of SLC25A25-AS1 in the serum of the patients with cervical cancer and the patients with cervical intraepithelial neoplasia (CIN) and explored the mechanism of SLC25A25-AS1 in the development of cervical cancer. The expression levels of SLC25A25-AS1 in the serum of normal controls‚ patients with cervical cancer‚ and patients with CIN were detected by RT-qPCR. The important role of SLC25A25-AS1 in HeLa cells was analyzed in vitro and in vivo experiments. Compared with the normal group‚ the expression level of serum SLC25A25-AS1 was decreased in patients with cervical cancer. In vitro‚ overexpression of SLC25A25-AS1 inhibited cell proliferation‚ migration‚ and invasion of HeLa cells. Tumor formation in nude mice assay showed that the subcutaneous tumor weight and volume of nude mice injected with SLC25A25-AS1-overexpressed HeLa cells were significantly smaller than that of nude mice injected with control cells (P<0. 05). SLC25A25-AS1 may play an important role in the development of cervical cancer and may serve as a new therapeutic target for cervical cancer.

The Gut Microbiota Changed by Short-term Low-dose Ethanol Withdrawal and Related to the Anxiety like Behavior in Male Rats / 中国生物化学与分子生物学报

Long-term regular ethanol intake could change the gut microbiota and affect anxiety and depression-like behaviors․ It is not clear that whether ethanol withdrawal after short-term low-dose drinking has an effect on the gut microbiota or whether it is related to anxiety-like behaviors․ In this study, 30 male Sprague￣Dawley rats were randomly divided into three groups: Ethanol-C: ethanol treatment group, treated with (5 g / kg, 25% V / V) ethanol for 14 days; Ethanol-2: ethanol withdrawal group, treated with (5 g / kg, 25% V / V) ethanol for 14 days and then withdrawal for one day; Ethanol-0: Control group, the rats were given the same amount of distilled water for 14 days․ Feces were collected from all rats, and high-throughput sequencing methods were used to analyze the effect of ethanol withdrawal after short-term low-dose drinking on the gut microbiota․ The open-field test and elevated plus-maze test were used to determine anxiety-like behaviors, and analyze the correlation between gut microbes and anxiety-like behaviors caused by alcohol withdrawal․ Taxonomic analysis of gut microbiota found that the composition and abundance in the ethanol withdrawal group were significantly different from those in the control group and the alcohol-treated group․ The Alpha diversity of gut microbiota in the ethanol withdrawal group was not significantly different from the control group and the ethanol treatment group, whereas the microbial community structure was significantly different․ The percentage of time spent in the open arms and total distance of rats in the ethanol withdrawal group were significantly reduced (P < 0․ 05) , and behavioral parameters were significantly positive correlated with Bacteroides, Fusobacterium, and Escherichia-Shigella (P < 0․ 05) , but significantly negative correlated with Rumenococcus, Trichospirillum and other bacteria (P < 0․ 05) ․ This study suggests that short-term low-dose ethanol withdrawal does not affect the Alpha diversity, but can change the abundance and community structure of the gut microbiota in rats, and the gut microbiota are correlated with anxiety-like behaviors in rats․ This study clarified the changes of gut microbiota after short-term low-dose ethanol withdrawal, and provided a new direction for the study of anxiety-like behaviors caused by short-term low-dose ethanol withdrawal․

Clinical anatomical study on the treatment of carpal tunnel syndrome with classic Acupotomy / 中国骨伤

OBJECTIVE@#To explore the safety of classic Acupotomy in the treatment of carpal tunnel syndrome.@*METHODS@#Twenty six adult specimens (15 males and 11 females), aged 60 to 95(82.54±6.94) years old, were selected from 10% formalin antiseptic fixation. There were 52 sides(two of them could not be tested). The study period was from November 2017 to May 2018. The specimens were collected from the body donation center of the school of basic medicine, Peking University. The operation of releasing the transverse carpal ligament on the human body specimen was simulated by the classic acupotomy, and the distance from the four points to the surrounding anatomical structure was measured to calculate the direct injury rate to the nerve and blood vessels, and the shortest distance between the acupotomy and the nerve and blood vessels was defined as ≥2 mm as safety.@*RESULTS@#In the experimental operation, the direct injury rate of nerve and blood vessel was 14% and 12% respectively. There was significant difference in the rate of direct nerve injury between the four injection points (0.05). Among the four points, there was a statistically significant difference in the safety of nerves(<0.05), and the safety of point 1 and point 3 of radial injection was higher than that of point 2 and point 4 of ulnar injection(<0.05). There was significant difference in the safety of blood vessels between the four points(<0.05), and the safety of radial point 1 was higher than that of ulnar point 2 and point 4 (<0.05).@*CONCLUSION@#The safety of the classic Acupotomy for carpal tunnel syndrome is related to the location of the needle entry point, and the safety of theradial proximal end of the needle is the highest.

Effects of estrogen receptor antagonist on the expressions of Erα, Erβ and p57kip2 in JEC cells of human endometrial carcinoma / 临床与实验病理学杂志

Purpose To explore the effects of estrogen receptor antagonist on the expression of estrogen receptor subtype (ERα, ERβ), and p57kip2 protein in human endometrioid carcinoma cells named JEC. Methods The JEC cells (moderately differentiated EC cells) cultured in vitro were treated with β-Estradiol (E2) (10～6 mol/L) and two types of estrogen receptor antagonists, tamoxifen (TAM) and fulvestrant (ICI182780) (10-6 mol/L). After 24, 48, 72 h, MTT was used to detect the growth condition of JEC cells, and the light microscopy and electron microscopy were used to observe the growth condition and morphological changes of cells, Western blot was used to detect the expression of ERα, ERβ, PR-A, PR-B and P57kip2 protein in JEC cells. Results MTT results: Compared with the control group, E2 could promote the proliferation of JEC cells significantly (P＜0.05), and ICI182780 could inhibit the proliferation of JEC cells obviously (P＜0.05). Compared with the E2 group, the proliferation ability of JEC cells in E2 + ICI182780 group were lower(P＜0.05). Morphological change: Compared with the control group, the cells density of E2 group increased obviously, and the pathologic mitosis was easy to seen in some cells. The cells density decreased obviously in ICI182780 group. Compared with E2 group, the cells density of E2 + TAM group and E2 + ICI182780 group were decreased, and pathological mitotic figures were difficult to seen. Western blot results: Compared with the control group, the expression of ERβ protein increased, and the expression of p57kip2 protein decreased in E2 group (P＜0.05). The expression of ERβ protein decreased, and the expression of p57kip2 protein increased in ICI182780 group and TAM group, and the difference was statistically significant between ICI182780 group and control group (P＜0.05). Compared with the E2 group, the expression of ERβ protein decreased, and the expression of p57kip2 protein increased in E2 + ICI182780 group and E2 + TAM group, and the difference was statistically significant between E2 + ICI182780 group and E2 group (P＜0.05). ERa protein of JEC cells did not expressed in experimental group or control group. Conclusion ERa protein are not expressed in JEC cells. ICI182780 have a stronger role in antagonizing estrogen, and may induce the expression of p57kip2 protein by down-regulating the expression of ERβ protein in JEC cells, block the cell cycle progression and inhibit the growth of tumor cells. TAM has a weaker estrogen like effect on the growth of JEC cells. It is possible that combined detection of the expression of ERa and p57kip2 protein in EC has an important reference value for individualized selection of endocrine therapy for EC patients.

A review on health inequ ality and related facot rs of the globa l elderly / 中国卫生政策研究

The rapidly aging population has attracted extensive attention to the health of elderly people .The elderly are a vulnerable group in society .Their adverse physical and mental health not only affects their own well-be-ing but also brings a heavy burden on families and society .Numerous studies have found that health inequality of the elderly is a problem throughout the world, and differences in gender, age, marital status, socioeconomic status, and social capital among the elderly are all contributing factors .This paper summarizes these factors through a literature review.We suggest that cross-sectional and longitudinal data should be combined and suitable indicators and mathe -matical models to Chinese conditions should be established in order to analyze the mechanisms of various factors and their effects on health inequality and serve as a reference for promoting the overall health of the elderly .

Effects of endogenous carbon monoxide on gene expression profiles associated with the apoptosis of pulmonary arterial smooth muscle cells / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To identify the gene expression profiles associated with the apoptosis of pulmonary arterial smooth muscle cells stimulated by carbon monoxide (CO).</p><p><b>METHODS</b>Primary cultured Sprague-Dawley rat pulmonary arterial smooth muscle cells (PASMC) were stimulated by platelet-derived growth factor (PDGF, 20 ng/mL) and hemin (20 μmol/L). Cells were harvested after 2 hrs and Affymetrix microarrays were used to detect the gene expression profile.</p><p><b>RESULTS</b>Some genes associated with Map2k3 (P38) signal pathway, such as CyclinD1, CyclinH, CyclinL1, MAP2K3, Kras and Nras, were upregulated, but P27 expression was downregulated after PDGF treatment. After endogenous CO treatment, some genes associated with P53 pathway, such as Gadd45α, P21 and Trp53inp1, were upregulated.</p><p><b>CONCLUSIONS</b>P53 pathway probably plays an important role in apoptosis of pulmonary arterial smooth muscle cells treated with endogenous CO.</p>

Study on GATA-6 Gene on Inhibition of Endogenous Carbon Monoxide on Pulmonary Vascular Smooth Muscle Cells Proliferation / 实用儿科临床杂志

Objective To investigate the effect of GATA-6 on endogenous carbon monoxide(CO) inhibited pulmonary vascular smooth muscle cells(PVSMCs) proliferation induced by platelet-derived growth factor(PDGF).Methods Tissue mass culture was done to get PVSMCs artery in SD rats.The PVSMCs were stimulated to proliferation by PDGF(20 ?g/L),and the 3 different concentrations of hemin[a substrate and inducer of heme oxygenase-1(HO-1)] add into the cultures to induce CO production.The PVSMC cell proli-feration was detected by 3-(4,5-dimethyl-2-thiazoly1)-2,5-diphenyl-2H-tetrazolium bromide(MTT) method,DNA synthesis was detected by thymidine incorporation,and GATA-6 mRNA expression was detected by reverse transcription polymerase chain reaction(RT-PCR).Results CO inhibited the PVSMCs proliferation induced by PDGF in a dose-dependent and time-dependent manner.Hemin with high concentration markedly inhibited the proliferation and DNA synthesis of PVSMCs.After 2 hours with PDGF,the expression of(GATA-6) mRNA markedly down-regulated,and began returned after 6 hours.However,CO could reversed this down-regulation.Conclusions CO can inhibit the proliferation of PVSMCs induced by PDGF.PDGF can result in the expression of GATA-6 mRNA down-regulated;the down-regulation is reversed by CO.This study suggested that CO maybe inhibit PVSMCs proliferation by regulating expression of GATA-6.