Clinicopathological features and prognosis of breast cancer with human epidermal growth factor receptor 2 low expression / 北京大学学报(医学版)

OBJECTIVE@#There is an increasing interest in human epidermal growth factor receptor 2 (HER2) low expression breast cancer with the result of novel anti-HER2 antibody-drug conjugates for breast cancer. HER2 low expression breast cancer is expected to become a new type of breast cancer. This study analyzed and compared the clinicopathological features and survival data of breast cancer with HER2 low expression group [immunohistochemistry (IHC) 1+ or IHC 2+, and fluorescence in situ hybridization (FISH) negative] and HER2 zero expression group (IHC 0), in order to explore the difference in clinical biology of HER2 low expression breast cancers.@*METHODS@#Among 1 250 female patients with primary non-metastatic breast cancer admitted to the Breast Disease Center of Peking University First Hospital from January 2014 to December 2017, 969 cases were HER2 negative (IHC 0, 1+, 2+, and FISH was not amplified). The clinicopathologic features and prognosis of the patients with HER2 low expression (IHC 1+ or 2+, and unamplified by FISH) and HER2 zero expression (IHC 0) were analyzed. Disease free survival (DFS) and overall survival (OS) were evaluated, survival rates were calculated by Kaplan-Meier curve, and survival differences were compared by Log-rank test. Cox regression analysis of univariate and multivariate prognostic factors. Bilateral test was used, and P < 0.05 was considered statistically significant.@*RESULTS@#In the 969 patients with HER2 negative breast cancer, 606 had HER2 low expression (62.54%) and 363 had HER2 zero expression (37.46%). Compared with breast cancer with HER2 zero expression, those with HER2 low expression had higher N stage (P=0.001) and TNM stage (P=0.044), the proportion of non-specific histological types was higher (82.7% vs. 79.1%, P=0.009), the histological grade was higher (P=0.048), and the positive rate of hormone receptor was higher (83.2% vs. 75.2%, P=0.003). The percentage of Ki-67 value index >30% was lower (30.4% vs. 36.6%, P=0.044). There was no significant difference in DFS and OS between the two groups (P>0.05). In the 969 cases, 777 were hormone receptor positive and 192 were hormone receptor negative (triple negative cancer). Among the 777 cases with hormone receptor positive, 504 (64.9%) were HER2 low expression, and 273 (35.1%) were HER2 zero expression. Compared with breast cancer with HER2 zero expression group, the HER2 low expression group had a younger age (P=0.016), a higher proportion of premenopausal patients (P=0.029), more lymph node involvement (P=0.002), and a higher total TNM stage (P=0.031), and less frequent histological types of lobular and mucinous carcinoma (3.6% vs. 7.3%, 4.8% vs. 10.6%, P=0.001). There was no difference in DFS and OS between HER2 low expression and zero expression (P>0.05). Among 192 patients with hormone receptor negative, there were 102 cases (53.1%) with HER2 low expression and 90 cases (46.9%) with HER2 zero expression. Compared with the HER2 zero expression groups, HER2 low expression group was older (P=0.001), the proportion of premenopausal patients was low (P=0.029), the histological grade was lower (P < 0.001), the Ki-67 value index was lower (P < 0.001), and androgen receptor positive rate was higher (58.8% vs. 34.4%, P < 0.001). DFS was better than HER2 zero expression group (P=0.038), but there was no difference in OS between the two groups (P>0.05).@*CONCLUSION@#HER2 low expression breast cancer accounts for about half of all breast cancers, and the incidence is much higher than that of HER2 positive breast cancer. Its clinicopathologic features are heterogeneous, and the status of hormone receptor expression has an impact on the clinical biology of this group.

Establishment and preliminary application of quantitative real-time PCR assay for the detection of SARS-CoV-2 subgenomic nucleocapsid RNA / 中华预防医学杂志

Objective: To establish a rapid and specific quantitative real-time PCR (qPCR) method for the detection of SARS-CoV-2 subgenomic nucleocapsid RNA (SgN) in patients with COVID-19 or environmental samples. Methods: The qPCR assay was established by designing specific primers and TaqMan probe based on the SARS-CoV-2 genomic sequence in Global Initiative of Sharing All Influenza Data (GISAID) database. The reaction conditions were optimized by using different annealing temperature, different primers and probe concentrations and the standard curve was established. Further, the specificity, sensitivity and repeatability were also assessed. The established SgN and genomic RNA (gRNA) qPCR assays were both applied to detect 21 environmental samples and 351 clinical samples containing 48 recovered patients. In the specimens with both positive gRNA and positive SgN, 25 specimens were inoculated on cells. Results: The primers and probes of SgN had good specificity for SARS-CoV-2. The minimum detection limit of the preliminarily established qPCR detection method for SgN was 1.5×102 copies/ml, with a coefficient of variation less than 1%. The positive rate of gRNA in 372 samples was 97.04% (361/372). The positive rates of SgN in positive environmental samples and positive clinical samples were 36.84% (7/19) and 49.42% (169/342), respectively. The positive rate and copy number of SgN in Wild strain were lower than those of SARS-CoV-2 Delta strain. Among the 25 SgN positive samples, 12 samples within 5 days of sampling time were all isolated with virus; 13 samples sampled for more than 12 days had no cytopathic effect. Conclusion: A qPCR method for the detection of SARS-CoV-2 SgN has been successfully established. The sensitivity, specificity and repeatability of this method are good.

Schisandrin B Improves the Hypothermic Preservation of Celsior Solution in Human Umbilical Cord Mesenchymal Stem Cells

BACKGROUND@#Human umbilical cord mesenchymal stem cells (hUCMSCs) have emerged as promising therapy for immune and inflammatory diseases. However, how to maintain the activity and unique properties during cold storage and transportation is one of the key factors affecting the therapeutic efficiency of hUCMSCs. Schisandrin B (SchB) has many functions in cell protection as a natural medicine. In this study, we investigated the protective effects of SchB on the hypothermic preservation of hUCMSCs. @*METHODS@#hUCMSCs were isolated from Wharton’s jelly. Subsequently, hUCMSCs were exposed to cold storage (4 °C) and 24-h re-warming. After that, cells viability, surface markers, immunomodulatory effects, reactive oxygen species (ROS), mitochondrial integrity, apoptosis-related and antioxidant proteins expression level were evaluated. @*RESULTS@#SchB significantly alleviated the cells injury and maintained unique properties such as differentiation potential, level of surface markers and immunomodulatory effects of hUCMSCs. The protective effects of SchB on hUCMSCs after hypothermic storage seemed associated with its inhibition of apoptosis and the anti-oxidative stress effect mediated by nuclear factor erythroid 2–related factor 2 signaling. @*CONCLUSION@#These results demonstrate SchB could be used as an agent for hypothermic preservation of hUCMSCs.

A study on the single -tube multiplex RT -real time PCR assay in detecting rotavirus,astrovirus and hepatitis A virus / 预防医学

Objective To develop and evaluate a single -tube multiplex RT -real time PCR assay for the simultaneous detection of rotavirus,astrovirus and hepatitis A virus.Methods Gen -bank sequences of rotavirus,astrovirus and hepatitis A virus were included as reference sequences.Primers and probes were designed based on the reference sequences.A multiplex real -time RT -PCR assay was developed,and the reproducibility,specificity and sensitivity of the assay were evaluated.Fecal samples from 1 28 patients with viral diarrhea were detected and verified by gene sequencing.Results There was high reproducibility and specificity of the single -tube multiplex real -time RT -PCR assay for detecting rotavirus,astrovirus and hepatitis A virus.Detection limits of 1 01 copies were achieved in tests of rotavirus,1 02 copies for astrovirus and hepatitis A virus in one reaction.3.1 3% and 1 7.97% of 1 28 clinical specimens were detected positive for astrovirus RNA and rotavirus RNA respectively.And the positive samples were verified by gene sequencing.Conclusion Rotavirus,astrovirus and hepatitis A virus can be detected and identified by the single -tube multiplex RT -real time PCR assay with high specificity and sensitivity.The assay developed in this study can be applied to the clinical diagnosis and epidemiological investigation.

Salidroside via ERK1/2 and PI3K/AKT/mTOR signal pathway induces mouse bone marrow mesenchymal stem cells differentiation into neural cells / 药学学报

To investigate the role of the extracellular signal-regulated kinase (ERK1/2) and PI3K/AKT/ mTOR signal pathway inducing bone marrow mesenchymal stem cells (BMSCs) differentiation into neural cells, mouse bone marrow-derived mesenchymal stem cell lines D1 cells were used as research object. And they were divided into control groups and salidroside (SD) groups. Different concentrations (5, 25, 50, 100 and 200 microg x mL(-1) of SD were used and SD (100 microg x mL(-1)) was used to induce at different time (0.5, 1, 3, 6, 9, 12, 24, 48 and 72 h). The immunofluorescence staining chemical technology, real-time PCR and Western blotting were used to detect the positive rates of NSE, MAP2, beta-Tubulin III, NES, GFAP and the expression levels of beta-Tubulin III, NSE, ERK1/2, AKT. The expression of ERK1/2 and NSE was detected when the ERK1/2 and PI3K/AKT/ mTOR signal pathway was blocked by PD98059 and LY294002. It indicated that the positive rates of NSE, MAP2, beta-Tubulin III, NES and GFAP were gradually enhanced with time increased. The expression level of NSE and beta-Tubulin III protein were significantly higher than those in control groups (P < 0.01). The expression of ERK1/2, AKT mRNA and protein were higher with concentration and time increased. When the ERK1/2 and PI3K/AKT/mTOR signal pathway were blocked, the expression levels of NSE, NES and beta-Tubulin III mRNA and NSE protein were inhibited significantly. It points out that SD can stimulate the ERK1/2 and PI3K/AKT/mTOR signal pathway to promote BMSCs differentiation into neural cells.

Effect of high-dose dexamethasone on BAFF and Tregs in patients with immune thrombocytopenic purpura / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate the change of B-cell activating factor of the TNF family (BAFF) and regulatory T-cells (Tregs) before and after high-dose dexamethasone(HD-DXM) therapy and assess the effect of BAFF on Treg cells in immune thrombocytopenic purpura (ITP).</p><p><b>METHODS</b>The plasma BAFF concentration was measured by ELISA, and Treg cell numbers by flow cytometry.</p><p><b>RESULTS</b>The plasma BAFF level \[(599.70 +/- 199.40) pg/ml\] was significantly increased (P < 0.05), and the percentage of Treg cells \[(1.56 +/- 0.73)%\] was significantly decreased (P < 0.01) in ITP patients before treatment as compared with that in controls \[(454.5 +/- 132.5) pg/ml and (4.08 +/- 1.08)%, respectively\]. After treatment with HD-DXM, the plasma BAFF level \[(296.9 +/- 119.7) pg/ml\] was significantly decreased (P < 0.01), and the percentage of Treg cells \[(5.94 +/- 2.22)%\] was significantly increased (P < 0.01). The BAFF level and Treg proportion had no significant correlation with platelets count (P > 0.05). In in vitro assays, no difference was found in the number of Treg cells between rhBAFF0 group and rhBAFF20 group \[(1.53 +/- 0.69)%, (1.49 +/- 0.67)%, P = 0.89)\].</p><p><b>CONCLUSION</b>BAFF level was increased and Treg cells decreased in ITP patients. HD-DXM might play a role in ITP treatment by down-regulating BAFF expression and up-regulating Treg cells number. BAFF had no influence on the number of Treg cells.</p>

Laparoscopic transhiatal extended gastrectomy for type II, III esophagogastric junction cancer: a preliminary report of 55 cases / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore the feasibility and safety of laparoscopic extended gastrectomy through the transhiatal approach in patients with esophagogastric junction cancer.</p><p><b>METHODS</b>From Feb 2008 to May 2010, 55 cases with Siewert type II or III esophagogastric junction cancer underwent laparoscopic transhiatal extended gastrectomy at the West China hospital. Clinical data were analyzed retrospectively.</p><p><b>RESULTS</b>Esophagogastric junction cancer was Siewert type II in 36 patients and Siewert type III in 19. Thirty-five cases underwent proximal gastrectomy, 20 total gastrectomy. There were 53 D2 lymph node excisions and 2 palliative resections. Fifty patients underwent laparoscopic extended gastrectomy successfully, with 5 converted to open operations. A safe anastomosis between inferior pulmonary vein and pulmonary hilum was achieved in the majority of patients. The mean operative time was(236.2±35.5) min and the mean estimated blood loss was(60.6±33.9) ml. There were no postoperative mortalities or anastomotic leakage/stenosis. No reoperations were required. Pleural laceration occurred in 11 cases during operation, of whom 10 were repaired intraoperatively and one was managed with drainage postoperatively. There were 3 patients developed pulmonary infection and one wound infection. Postoperative recovery was uneventful in other patients.</p><p><b>CONCLUSION</b>Laparoscopic transhiatal extended gastrectomy is feasible and safe for patients with esophagogastric junction cancer.</p>

The role of interleukin-18 and interleukin-18 receptor in predominant Th1 immune response of patients with immune thrombocytopenia / 中华血液学杂志

<p><b>OBJECTIVE</b>To evaluate the role of interleukin (IL)-18 and IL-18 receptor (IL-18R) in the predominant Th1 type cytokine response in patients with immune thrombocytopenia (ITP).</p><p><b>METHODS</b>Fifteen patients with active phase ITP, eighteen in remission and thirteen healthy controls were enrolled in this study. T-bet and GATA-3 mRNA levels in peripheral blood mononucleated cells (PBMNC) were measured by reverse transcriptase polymerase chain reaction (RT-PCR); the plasma IL-18 level by enzyme linked immunosorbent assay (ELISA), the expression of IL-18R on CD3(+) lymphocytes and total lymphocytes by flow cytometry(FCM).</p><p><b>RESULTS</b>The T-bet mRNA levels in patients with active phase ITP was 3.572 fold as much as that in the controls (P < 0.05), while the GATA-3 mRNA levels were 0.378 fold of that in controls (P < 0.05). The levels of plasma IL-18 and IL-18R on CD3(+) lymphocytes were significantly increased in active phase ITP than in remission phase and controls. There was no difference in ratio of T-bet/GATA-3 between remitted ITP and controls and so was for T-bet mRNA, GATA-3 mRNA, plasma IL-18 and IL-18R on CD3(+) lymphocytes.</p><p><b>CONCLUSION</b>ITP as a disease of Th1-dominant response there is an unbalance between T-bet and GATA-3 in its active phase; IL-18 and IL-18R being upregulated.</p>

Screening and identification of human anti-c-Met Fab from a phage antibody library / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To screen anti-c-Met Fab from a phage antibody library and identify its binding activity.</p><p><b>METHODS</b>The expression of c-Met of HCC lines was identified by Western blot and immunofluorescence. Antibodies against c-Met were screened with immobilized antigen. After five rounds of panning, 30 randomly selected clones were identified by phage ELISA to select specific clones with high affinity. The positive clones were selected for Fab soluble expression in TOP10F and the binding activities were analysed in HCC lines.</p><p><b>RESULTS</b>c-Met expressed in HCC membrane was confirmed by Western blot and immunofluorescence. A Fab fragment named AM2-26 with fine activity to c-Met was selected. AM2-26 binding specificity was confirmed by IP, FACS and immunofluorescence.</p><p><b>CONCLUSION</b>The anti-c-Met Fab binding to c-Met in HCC provides a promising candidate for the biotherapy of hepatoma.</p>

Effects of different dialysates on apoptosis and expression of PKC? of U937 cell line / 上海第二医科大学学报

Objective To investigate the effects of different dialysates on expression of protein kinase C-? (PKC?) and apoptosis of U937 cell line. Methods Different dialysates were added into culture fluid with U937 cell line at exponential phase of growth, and groups were divided: fluid A+fluid B group (dialysate A+dialysate B), fluid A+fluid B+rottlerin (PKC? specific inhibitor)group, fluid A+powder B group (dialysate A+powder B) and fluid A+powder B + rottlerin group. Besides, blank control group and normal control group were established. Cells were harvested 24 h and 48 h after treatment, morphological changes were observed by Hoechst33258 fluorescence staining, cell apoptosis was measured by Annexin-V-FITC/PI double staining, and expression of PKC? mRNA and protein was detected by RT-PCR and Western blotting, respectively. Results Cell apoptosis significantly increased in fluid A+powder B group, with typical morphology of apoptosis. After treatment for 24 h and 48 h, cell apoptosis rates in fluid A+powder B group were significantly higher than those at corresponding time points in blank control group, normal control group and fluid A+powder B+rottlerin group (P0.05). Conclusion Fluid A+powder B can significantly increase apoptosis of U937 cell line, the mechanism of which may be associated with the up-regulation of expression of PKC?. Compared with fluid A+powder B, fluid A+fluid B is superior in reducing apoptosis of peripheral blood monouclear cells.