Gene mutations of esophageal squamous cell carcinoma based on next-generation sequencing / 中华医学杂志(英文版)

BACKGROUND@#Esophageal squamous cell carcinoma (ESCC) is one of the most aggressive cancers without effective therapy. To explore potential molecular targets in ESCC, we quantified the mutation spectrum and explored the relationship between gene mutation and clinicopathological characteristics and programmed death-ligand 1 (PD-L1) expression.@*METHODS@#Between 2015 and 2019, 29 surgically resected ESCC tissues and adjacent normal tissues from the Fourth Hospital of Hebei Medical University were subjected to targeted next-generation sequencing. The expression levels of PD-L1 were detected by immunohistochemistry. Mutational signatures were extracted from the mutation count matrix by using non-negative matrix factorization. The relationship between detected genomic alterations and clinicopathological characteristics and PD-L1 expression was estimated by Spearman rank correlation analysis.@*RESULTS@#The most frequently mutated gene was TP53 (96.6%, 28/29), followed by NOTCH1 (27.6%, 8/29), EP300 (17.2%, 5/29), and KMT2C (17.2%, 5/29). The most frequently copy number amplified and deleted genes were CCND1/FGF3/FGF4/FGF19 (41.4%, 12/29) and CDKN2A/2B (10.3%, 3/29). By quantifying the contribution of the mutational signatures to the mutation spectrum, we found that the contribution of signature 1, signature 2, signature 10, signature 12, signature 13, and signature 17 was relatively high. Further analysis revealed genetic variants associated with cell cycle, chromatin modification, Notch, and Janus kinase-signal transducer and activator of transcription signaling pathways, which may be key pathways in the development and progression of ESCC. Evaluation of PD-L1 expression in samples showed that 13.8% (4/29) of samples had tumor proportion score ≥1%. 17.2% (5/29) of patients had tumor mutation burden (TMB) above 10 mut/Mb. All samples exhibited microsatellite stability. TMB was significantly associated with lymph node metastasis (r = 0.468, P = 0.010), but not significantly associated with PD-L1 expression (r = 0.246, P = 0.198). There was no significant correlation between PD-L1 expression and detected gene mutations (all P > 0.05).@*CONCLUSION@#Our research initially constructed gene mutation profile related to surgically resected ESCC in high-incidence areas to explore the mechanism underlying ESCC development and potential therapeutic targets.

Association of Overlapped and Un-overlapped Comorbidities with COVID-19 Severity and Treatment Outcomes: A Retrospective Cohort Study from Nine Provinces in China / 生物医学与环境科学（英文）

Objective@#Several COVID-19 patients have overlapping comorbidities. The independent role of each component contributing to the risk of COVID-19 is unknown, and how some non-cardiometabolic comorbidities affect the risk of COVID-19 remains unclear.@*Methods@#A retrospective follow-up design was adopted. A total of 1,160 laboratory-confirmed patients were enrolled from nine provinces in China. Data on comorbidities were obtained from the patients' medical records. Multivariable logistic regression models were used to estimate the odds ratio ( @*Results@#Overall, 158 (13.6%) patients were diagnosed with severe illness and 32 (2.7%) had unfavorable outcomes. Hypertension (2.87, 1.30-6.32), type 2 diabetes (T2DM) (3.57, 2.32-5.49), cardiovascular disease (CVD) (3.78, 1.81-7.89), fatty liver disease (7.53, 1.96-28.96), hyperlipidemia (2.15, 1.26-3.67), other lung diseases (6.00, 3.01-11.96), and electrolyte imbalance (10.40, 3.00-26.10) were independently linked to increased odds of being severely ill. T2DM (6.07, 2.89-12.75), CVD (8.47, 6.03-11.89), and electrolyte imbalance (19.44, 11.47-32.96) were also strong predictors of unfavorable outcomes. Women with comorbidities were more likely to have severe disease on admission (5.46, 3.25-9.19), while men with comorbidities were more likely to have unfavorable treatment outcomes (6.58, 1.46-29.64) within two weeks.@*Conclusion@#Besides hypertension, diabetes, and CVD, fatty liver disease, hyperlipidemia, other lung diseases, and electrolyte imbalance were independent risk factors for COVID-19 severity and poor treatment outcome. Women with comorbidities were more likely to have severe disease, while men with comorbidities were more likely to have unfavorable treatment outcomes.

Current situation and reflection of preserving reproductive function in cervical cancer patients / 中国实用妇科与产科杂志

With the standardization and popularization of cervical cancer screening techniques,more and more young patients with cervical cancer are being detected at an early stage,and most patients with cervical cancer at reproductive age have a strong intention to preserve fertility.This article will summarize the operation indication,operation methods,safety,oncologic outcomes and related controversies of fertility preservation surgery in patients with cervical cancer.

Prevention of chemotherapy-induced ovarian damage in patients with gynecological malignant tumors / 中国实用妇科与产科杂志

Improvement in diagnosis has made it possible to have the early detection of gynecologic malignancies.Among premenopausal women,the loss of future fertility or ovarian function is considered among the most dreadful long-term side effects of treatments.Therefore,the preservation of fertility and ovarian function during chemotherapy treatment is of great important.This article will summarize the effect of chemotherapy on ovarian function and protective measures for chemotherapy-induced ovarian damage.

Limited Role of Promoter Methylation of and in Triage of Low-Grade Squamous Intraepithelial Lesion / 中华医学杂志(英文版)

<p><b>Background</b>Promoter methylation of MGMT and C13ORF18 has been confirmed as a potential biomarker for early diagnosis of cervical cancer. The aim of this study was to evaluate the performance of MGMT and C13ORF18 promoter methylation for triage of cytology screening samples and explore the potential mechanism.</p><p><b>Methods</b>Methylation-sensitive high-resolution melting was used to detect promoter methylation of MGMT and C13ORF18 in 124 cervical samples. High-risk human papillomavirus (HR-HPV) was detected by the Digene Hybrid Capture 2. Gene methylation frequencies in relation to cervical intraepithelial neoplasia (CIN) were analyzed. Frequencies were compared by Chi-square tests. The expression of gene biomarkers and methylation regulators was analyzed by immunohistochemical staining, real-time fluorescence quantitative polymerase chain reaction, and Western blot.</p><p><b>Results</b>For triage of low-grade squamous intraepithelial lesion (LSIL), gene methylation increased specificity from 4.0% of HR-HPV detection to 30.8% of MGMT (χ = 9.873, P = 0.002) and to 50.0% of C13ORF18 (χ = 21.814, P = 0.001). For triage of atypical squamous cells of undetermined significance, HR-HPV detection had higher positive predictive value of 54.8%. Either MGMT or C13ORF18 methylation combined with HR-HPV increased the negative predictive value to 100.0% (χ = 9.757, P = 0.002). There was no relationship between MGMT and C13ORF18 expression and DNA methylation (χ = 0.776, P = 0.379 and χ = 1.411, P = 0.235, respectively). MBD2 protein level in cervical cancer was relatively lower than normal cervical tissue (t = 4.11, P = 0.006).</p><p><b>Conclusions</b>HR-HPV detection is the cornerstone for triage setting of CIN. Promoter methylation of MGMT and C13ORF18 plays a limited role in triage of LSIL. Promoter methylation of both genes may not be the causes of gene silence.</p>

A comparative clinical study of postoperative intensity-modulated radiotherapy combined with concurrent and sequential chemotherapy for high-risk early-stage cervical cancer / 中华放射肿瘤学杂志

Objective To evaluate the efficacy and acute toxieities of postoperative intensity-modulated radiotherapy (IMRT) combined with concurrent (C-IMRCT) or sequential chemotherapy (S-IMRCT) in the treatment of high-risk early-stage cervical cancer.Methods A retrospective study was performed on the clinical data of 105 patients with high-risk early-stage (Ⅰ B1-Ⅱ A2) cervical cancer from 2009 to 2017.Those patients were divided into C-IMRCT group (n=73) and S-IMRCT group (n=32).The 5-year disease-free survival (DFS) and overall survival (OS) rates,recurrence rate,metastasis rate,and acute toxicities were compared between the two groups.The survival rates were calculated by the Kaplan-Meier method and analyzed by the log-rank test.Univariate prognostic analysis was performed by the log-rank test.Recurrence,metastasis,and adverse reactions were compared using continuous correction chi-square test.Results The median follow-up time was 20 and 23 months in the C-IMRCT group and the S-IMRCT group,respectively (P=0.813).There were no significant differences in the 5-year DFS and OS rates between the two groups (72.6％ vs.72.5％,P=0.918;82.8％ vs.78.5％,P=0.504).There were no significant differences in the recurrence and metastasis rates between the two groups (P=0.598;P=1.000).The univariate prognostic analysis showed that no pathological factor affected prognosis.There were no significant differences in the incidence rates of grade 1-2 hematological toxicity,diarrhea,and urinary tract infection between the two groups (46.6％ vs.41.9％,P=0.884;P=0.854;P=0.271).Conclusions Further clinical studies are needed in terms of the survival rate in patients with high-risk early-stage cervical cancer receiving C-IMRCT.

Mesoporous nano-bioactive glass microspheres as a drug delivery system of minocycline / 北京大学学报(医学版)

OBJECTIVE@#To construct mesoporous nano-bioactive glass (MNBG) microspheres load-release minocycline as an antibacterial drug delivery system.@*METHODS@#Sol-gel method was used to synthesze MNBG microspheres as drug carrier. The MNBG consisted of SiO2, CaO, and P2O5. According to the content of silicon, MNBG microspheres were divided into four groups (60S, 70S, 80S and 90S). Scanning electron microscopy (SEM) was used to observe the surface characteristic and particle size of MNBG; Nitrogen adsorption-desorption experiment was performed to calculate the MNBG's specific surface area and the pore sizes; The Fourier transform infrared spectrum (FT-IR) and the thermogravimetric analysis were conducted to calculate the loading efficiencies of minocycline hydrochloride; UV spectrophotometric was used to determine the cumulative release of minocycline from drug-loaded particles in PBS solution within 21 d. Agar diffusion test (ADT) was performed to evaluate the antibacterial properties on Enterococcus faecalis. The inhibition zone was observed and the diameter was measured.@*RESULTS@#The MNBG microspheres had good dispersion, large surface area, and even particle size. The pore sizes ranged from 4.77 nm to 7.33 nm. The loading experiment results showed that the minocycline hydrochloride loading efficiency of MNBG was related to the pore size of the microspheres. Among 60S, 70S, 80S and 90S, 60S MNBG had the highest loading efficiency of 16.33% due to its high calcium content and large pore sizes. A slow minocycline release rate from MNBG particles in PBS solution until d 21 was observed. It was showed that a burst release of 28% of the total drug for the first 24 h. A cumulative release of 35% was found, and the final concentration of minocycline maintained at about 47 mg/L. ADT showed that mino-MNBG had inhibitory effect on the growth of Enterococcus faecalis. 1 g/L minocycline, 1 g/L mino-MNBG, and 0.1 g/L minocycline presented inhibition zone, however, PBS and 1 g/L MNBG didn't. The diameter of the inhibition zone of minocycline groups was significant larger than that of mino-MNBG group (P<0.05), which was also significant larger than those of PBS and MNBG groups (P<0.05). It showed that mino-MNBG drug delivery system had antibacterial properties on Enterococcus faecalis.@*CONCLUSION@#The 60S MNBG that can effectively load and release minocycline may be an ideal drug carrier.

Down-regulation of HDAC6 Expression Promotes Apoptosis of Human Leukemia K562 Cells / 中国实验血液学杂志

OBJECTIVE@#To study the promoting-apoptosis effect of HDAC6 on the human leukemia cells and its mechanism.@*METHODS@#The siRNA interference technology was used to inhibit the expression of HDAC6 gene, the RT-PCR and Western blot were used to detect the expression of HDAC6 and related signal pathway proteins respectively, the flow cytometry and Hoechest staining were used to detect the apoptosis and morphology changes of K562 cells.@*RESULTS@#Compared with the periphal blood monocyte and bone marrow stromal cells of healthy volunteers, the expression level of HDAC6 in leukemia cell lines was up-regulated significantly(P<0.05); the flow cytometry and Hoechest staining showed that after interference of HDAC6 gene, the apoptosis of K562 cells increased, moreover the cell morphology was changed; the Western blot detection showed that the interfering HDAC6 increased BAX/BCL-2 ratio and cleaved caspase 3 expression, and activated the MAPK, ATK, ERK signaling pathway.@*CONCLUSION@#The interferance of HDAC6 can promote the K562 cell apoptosis, its mechanism may relate with activation of MAPK signaling pathway.

Dehydroepiandrosterone indirectly inhibits human osteoclastic resorption via activating osteoblastic viability by the MAPK pathway / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Dehydroepiandrosterone (DHEA) is widely known for its beneficial effect on postmenopausal osteoporosis, although the underlying mechanisms remain mainly unclear. In this study, we tried to determine the activation of mitogen-activated protein kinase signal pathways during DHEA treatment and the indirect role of osteoblasts (OBs) on osteoclasts under the DHEA treatment of postmenopausal osteoporosis.</p><p><b>METHODS</b>Primary human OBs and osteoclast-like cells were cultured and, we pretreated OBs with or without U0126 (a highly selective inhibitor of both MEK1 and MEK2). The OBs were treated with DHEA. We then tested the effects of DHEA on human osteoblastic viability, osteoprotegerin production and the expression of phosphor-ERK1/2 (extracellular signal-regulated kinase). In the presence or absence of OBs, the function of osteoclastic resorption upon DHEA treatment was calculated.</p><p><b>RESULTS</b>DHEA promoted the human osteoblastic proliferation and inhibited the osteoblastic apoptosis within the concentration range of 10(-8) - 10(-6) mol/L (P < 0.05, P < 0.01, respectively). Within the effective concentration range, the expression of phosphor-ERK1/2 and osteoprotegerin was increased by DHEA and blocked by U0126. In the presence of OBs, DHEA could significantly decrease the number and the area of bone resorption lacuna (P < 0.05 and P < 0.01, respectively). Without OBs, however, the effects of DHEA on the bone resorption lacuna were almost completely abolished.</p><p><b>CONCLUSIONS</b>DHEA could indirectly inhibit the human osteoclastic resorption through promoting the osteoblastic viability and osteoprotegerin production, which is mediated by mitogen-activated protein kinases signal pathway involving the phosphor-ERK1/2.</p>

Sodium nitrite induces epithelial-mesenchymal transition of SMMC-7721 cells / 药学学报

This study is to find out the induction by sodium nitrite of epithelial-mesenchymal transition (EMT) in human hepatocellular carcinoma cells, SMMC-7721. After treatment of SMMC-7721 with 0.25 - 25 mmol.L-1 sodium nitrite for 48 h, the assays used include enzyme-linked immunosorbent assay (ELISA) for evaluation of TGF-beta1, IL-6 and IL-8 level in the conditioned medium, phase-contrast microscopy for morphology observation, and scratch wound healing as well as transwell migration assays for measurement of migration and metastatic potential. Additionally, the hallmarks of EMT, p-AKT and its downstream signaling molecules were examined by Western blotting. The results showed that TGF-beta1 secreted by SMMC-7721 elevated significantly in a dose-dependent fashion, whereas the increased IL-8 and IL-6 did not show dose-dependent response. The EMT was induced by exposure of SMMC-7721 with 0.25 mmol.L-1 of sodium nitrite, which was characterized by increased level of Vimentin, decreased E-cadherin and elevated activity of migration and metastatic potential. The results suggest that sodium nitrite could induce SMMC-7721 EMT by increased secretion of TGF-beta1 and IL-8.

Predominant effect of bushen ningxin decoction on postmenopausal osteoporosis in mice / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the preventive and therapeutic effect of Bushen Ningxin decoction (BSNX) on postmenopausal osteoporosis.</p><p><b>METHODS</b>The BALB/c female mice postmenopausal osteoporosis model was established. The model mice were treated by BSNX, with 17beta-estradiol (E2) and normal saline as positive and negative control, respectively. All mice were sacrificed after 12 weeks' treatment, the serum cytokines Th1/Th2, bone mineral density (BMD) of vertebrae (L3 - 4) and left femur were determined, and morphological quantitative analysis of bone tissue of right femurs was performed and osteoprotegerin (OPG) mRNA expression in tibia was detected by semi-quantitative RT-PCR. The ratio in weight of uterus to body was calculated, and uterine slice was gotten for histological observation.</p><p><b>RESULTS</b>As compared with the negative control group, the level of interferon-gamma (IFN-gamma) was significantly increased (P < 0.01) and interleukin-4 (IL-4) decreased (P < 0.05) in the BSNX treated group. The BMD of mice were improved, area of bone trabecula and OPG mRNA expression were increased in the BSNX treated and E2 treated group (P < 0.01). But the uterus in the former was significantly smaller than that in the latter (P < 0.01), while it was not significantly different to that in the negative control group.</p><p><b>CONCLUSION</b>BSNX can selectively prevent and cure the postmenopausal osteoporosis, it has no or slight stimulation on uterus. The mechanism may relate with its effects in regulating the deviation of Th1/Th2, enhancing the OPG expression and inhibiting the activity of osteoclasts.</p>

Effect of bushen ningxin decoction on proliferation and apoptosis of murine osteoblast / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To investigate the effect of Bushen Ningxin decoction (BSNXD) on postmenopausal osteoporosis.</p><p><b>METHODS</b>Postmenopausal osteoporosis model of female BALB/c mice were founded and afforded with BSNXD. The experiment was ceased, and the skeleton and blood of mice in each group were collected following 12 weeks of the treatment. Change of osteoblastic organelle in right femur was observed by transmission electron microscope (TEM). Osteoblasts from calvaria of different groups were isolated and cultured by the tissue fragment migrating growth method and the proliferation ratio of osteoblasts in the second passage was examined with thiazolyl blue reagents when cultured for 1 day and 5 days, respectively.</p><p><b>RESULTS</b>TEM showed that the osteoblasts of the animal treated by BSNXD had more cell organelles and developed Golgi complex, whose ability of proliferation(P < 0.05) and anti-apoptosis is stronger than that of OVX group.</p><p><b>CONCLUSION</b>BSNXD can accelerate the proliferation and inhibit the apoptosis of osteoblasts, the effective concentration of pharmacological serum varies from 10% to 20%.</p>