Objective To observe the effect of fasudil on cytoskeleton reconstruction in mouse podocytes induced by angiotensin Ⅱ (Ang Ⅱ),as well as to study the protective mechanism of fasudil in the pathological changes of podocytes.Methods Conditionally immortalized mouse podocytes were treated with Ang Ⅱ (10-7 mol/L).The podocytes were pre-incubated for 30 min or 60 min with various concentrations of fasudil (10-8,10-7,10-5 mol/L),then Ang Ⅱ (10-7 mol/L) were added and furtherincubated for 24 hours.The cytoskeleton distribution of podocyte as indicated by F-actin and synaptopodin was observed by fluorescence microscopy and Western blotting.At the same time,the activity of Rho signal pathway that mediates actin filament polymerization was analyzed by measuring the extent of Rho-associated coiled-coil-containing protein kinase 1 (ROCK-1) and myosin phosphatase-1 (MYPT1).Results Compared to the control group,AngⅡ disrupted the podocyte actin cytoskeleton and significantly decreased the expression of synaptopodin (P ＜ 0.05),while fasudil stabilized the actin filaments,and improved the synaptopodin expression (P ＜ 0.05).The expression enhancement of ROCK-1 and MYPT1 by Ang Ⅱ were reduced significantly by fasudil (P＜0.05).Conclusion The cytoskeleton reconstruction of podocytes can be induced by Ang Ⅱ and inhibited by fasudil,which suggests that the protective effect of fasudil may be partially contributed to the Rho/ROCK signaling pathway.

Objective To research on a whole blood control material for lymphocyte subset counting by flow cytometry(FCM).Methods To detect lymphocyte subset in whole blood with different preservers by flow cytometric multi-color analysis.Results whole blood control material for lymphocyte subset counting by FCM was prepared.In 2-8℃ refrigerator, the light scatter and CD45 of leukocytes of whole blood control were stable in 72 days. The cluster of lymphocyte, monocyte, neutrophil in plot were separated easily from debris. The lymphocyte subset of whole blood control can be counted by FSC/SSC or CD45/SSC gating. The variation of lymphocyte subset count was less in different preserving day. The coefficient of variation (CV) of lymphocyte subset count was less than 6.5% in our laboratory and less than 13% in external quality assessment among 56 laboratories in China.Conclusion The whole blood control prepared by us can be used for internal quality control and external quality assessment in lymphocyte subset counting by FCM, it is very important signification to ensure the quality and accuracy of lymphocyte subset count.

Objective To explore the influence of honokiol on growth,cell cycle and radiosensitivity in nasopharyngeal carcinoma cells,CNE-1 and CNE-2.Methods MTT assay and clonogenic assay were used to detect cell growth and survival respectively.Flow cytometry was employed to analyze cell cycle progression.Annexin V-FITC kit was used to detect cell apoptosis.Western blot assay was applied to examine protein expression.Results Honokiol signficantly inhibited proliferation of CNE-1 and CNE-2 cells in a dose and time dependent manner,the IC50 value was 2.84 and 2.68 μmol/L(24 h)and 2.50 and 2.20 μmol/L (48 h),respectively.After being treated with 2.5 μmol/L honokiol for 24 h,the ratios of early apoptosis,late apoptosis and necrosis were 24.53％,23.05％ and 7.13％ in CNE-1 cells compared with the control group(t =-41.17,-8.18,-6.08,P ＜0.05).The expression levels of pro-apoptotic proteins Caspase-3 and Bax were significantly increased to 2.31 and 1.89 times (t =-15.92,-17.15,P ＜ 0.05),4.43 and 1.85 times (t =-29.39,-13.47,P ＜ 0.05).Simultaneously the expression level of anti-apoptotic protein Bcl-2 was reduced by 2.22 and 2.74 times(t =26.94,66.14,P ＜ 0.05) as compared with controls after being treated with 4 and 3 μmol/L honokiol.Additionally,honokiol at lower doses signicantly enhanced the senstivity of CNE-1 and CNE-2 cells to X-ray irradiation.The SER was 1.41 and 1.88 in CNE-1 and CNE-2 cells.3 Gy irradiation of X-rays increased the proportion at G2/M state in both cell lines (t =-14.96,-19.26,P ＜ 0.05).Honokiol reduced the G2/M cell cycle arrest induced by irradiation significantly (t =7.65,4.98,P ＜ 0.05).Simultaneously,cyclin B1 protein expression obviously elevated (t =-33.07,-73.49,P ＜ 0.05).Conclusions Honokiol is a potent inhibitor of nasopharyngeal carcinoma cell growth by inducing cell apoptosis and necrosis and works as a radiosensitizer by disrupting G2/M cell cycle checkpoint.

Objective To investigate the effect of EGCG on the radiosensitivity of human nasopharyngeal carcinoma CNE-1 cells.Methods CNE-1 cells were divided into four groups:control,EGCG treatment,UVC or X-ray exposure,and EGCG combined with UVC or X-rays.After treatment with different concentrations of EGCG for 24,48 and 72 h and UVC or X-rays,cell growth was determined with MTT assay,cell survival was measured with clonogenic assay,cell cycle was deteeted with flow cytometry,cell apoptosis was detected by the annexin V-FITC cell apoptosis kit,and protein expression was assayed by Westem blot.Results EGCG inhibited cell growth in a dose-and time-dependent manner(r =0.817and 0.364).Compared with UVC or X-ray irradiation alone,the radiosensitivity of CNE-1 cells was enhanced by 2 h pre-treatment of 50 μmol/L EGCG,which disrupted S phase arrest caused by UVC( t =18.68,P ＜ 0.05 ) and increased the population of S and G2/M arrest caused by X-rays ( t =7.11 and 6.99,P ＜0.05 ).UVC could cause a significant increase of sub-G1 population( t =6.67,P ＜ 0.05 ) and Annexin V-FITC assay indicated apoptosis was further elevated by EGCG ( t =10.28,P ＜ 0.05 ).However,no significant induction of apoptosis was observed in the cells either irradiated with X-rays alone or combinationly treated with EGCG and X-rays.The combination treatment of EGCG and UVC significantly increased the expression of Bax and Caspase-3 proteins,but failed to affect Bcl-2 protein expression.Conclusions EGCG enhances the growth inhibition of CNE-1 cells caused by UVC or X-rays,which is relevant to apoptosis induction or cell cycle arrest.

Objective To investigate the effect of omeprazole combined with FOLFOX scheme as an adjuvant therapy for stage Ⅱ or Ⅲ colon cancer patients after a radical resection.Methods 98 stage Ⅱ or Ⅲ colon cancer patients in our hospital from January 2008 to December 2009 were randomly divided into study group (48 cases) receiving regimen of omeprazole combined with FOLFOX and control group (50 cases) treated with FOLFOX chemotherapy after radical colectomy.Surgical specimens were examined for expression of V-ATPase protein.Chemotherapy period was 6 months,8-12 courses.We observed results of follow-up curative effect,comparing the side effects and postoperative 2 year,3 year and 5 year disease-free survival rate (DFS) difference using statistical analysis.Results Study was completed in all 93 cases,5 cases were lost to follow-up.The baseline data distribution in the two groups were balanced basically.In study group the gastrointestinal side effects of chemotherapy was lower than the control group (x2 =4.924 6,P =0.026).In the two groups,the 2-year,3-year and 5-year DFS were 73％ vs 60％ (x2 =1.743 7,P =0.187),62％ vs50％ (x2 =1.4075,P=0.235),49％ vs40％ (x2 =0.8159,P=0.366) (P＞0.05).V-ATPase protein expression was 71％ (70/98) in all samples.The 2-year and 3-year DFS of patients for V-ATPase protein positive expression in the two groups were 75％ vs 51％ (x2 =3.970 8,P =0.046),66％ vs 40％ (x2 =4.399 5,P =0.036).Compared with the control group,the 2-year,3-year DFS increased in the study group (P ＜ 0.05).In stage Ⅲ colon cancer patients,the 2-year DFS was 73％ vs 47％ (x2 =4.504 5,P =0.034).Conclusions PPI combined with FOLFOX in V-ATPase protein positive expression or Ⅲ stage colon cancer patients after radical colectemy improves long-term survival,as well as reduces the gastrointestinal side effects of chemotherapy.

COVID-19 has emerged around the world since December 2019 and rapidly evolves as a major global public health challenge. SARS-CoV-2 is a positive single-stranded RNA virus with a relatively high natural mutation rate. The emergence of SARS-CoV-2 variants is generally considered as the outcome of a “mutation-selection-adaption” process. The enzyme APOBECs that is activated in the inflammatory microenvironment may facilitate viral mutagenesis. Currently， the major SARS-CoV-2 variants that impose public health significance are B.1.1.7， B.1.351， P.1， B.1.6.7.2， B.1.1.529 and so on， with probably increasing transmissibility， decreasing reactivity to the neutralizing antibodies， immune escape， and enhancing disease severity and mortality. Consequently， it warrants up-to-date countermeasures against evolving SARS-CoV-2， including tracing viral mutagenesis， developing vaccines against major variants， and enhancing social distancing.

Sarcopenia is a systemic syndrome characterized by decreased muscle mass and muscle strength and decline of motor function.Sarcopenia affects quality of life and disease prognosis of patients seriously, because of the low awareness rate, low treatment rate and high cardiovascular risk. Patients with uremia undergoing peritoneal dialysis are likely to suffer from sarcopenia due to dietary restriction, loss of protein and high catabolism. This article summarizes the diagnostic methods, risk factors, predictive markers and intervention measures for sarcopenia in peritoneal dialysis patients.

Objective:To provide scientific basis for prevention and control measures of island regions through analyzing the characteristics of malignant tumor incidence and mortality in eastern island areas of China and comparing the data with the national cancer mortality data in the same period. Methods:Using the incidence and mortality data of malignant tumor in Daishan county,Zhoushan collected by the Daishan Center for Disease Control and Prevention(CDC)from 2014 to 2019,which was stratified by gender,the crude incidence rates(CIR)and crude mortality rates(CMR)were calculated,and the top 10 malignant tumors with the highest incidence or mortality rates were then ranked.The Segi's world standard population was used to calculate the age-standardized incidence(ASIR)and age-standardized mortality rate(ASMR).The local data were compared with the national cancer mortality data from 2014 to 2019,and Chi-square test was used to analyze the differences between these two sets of data using the SPSS software.The difference was considered statistically significant when P<0.05. Results:There was a total of 7 305 incidence cases of malignant tumors in Daishan county,Zhoushan from 2014 to 2019.The CIR was 662.39/105 and the ASIR was 306.81/105.Notably,the CIR was the highest in the age group of 75-79 years old.The top 5 malignant tumors with the highest incidence rates were lung cancer(27.15％),gastric cancer(12.76％),liver cancer(10.95％),colorectal cancer(6.92％)and breast cancer(5.42％),whose ASIR were 75.09/105,32.06/105,31.01/105,17.81/105 and 18.36/105,respectively.There was a total of 3 412 mortality cases of malignant tumors in Daishan county,Zhoushan from 2014 to 2019.The CMR was 309.39/105 and the ASMR was 122.73/105.Notably,the CMR was the highest in the age group of 80-85 years old.The top 5 malignant tumors with the highest mortality rates were lung cancer(24.94％),liver cancer(18.64％),gastric cancer(17.00％),colorectal cancer(7.56％)and esophageal cancer(5.72％),whose ASMR were 29.65/105,24.97/105,19.01/105,8.75/105 and 6.60/105,respectively.The total ASMR of malignant tumors in Daishan county,Zhoushan was higher than national total ASMR from 2014 to 2019(100.34/105)(P<0.001).Specifically,the ASMR of gastric cancer,lung cancer and colorectal cancer in Daishan county were significantly higher than national levels(gastric cancer:12.46/105;liver cancer:16.45/105;colorectal cancer:6.63/105)(P<0.01),whereas no significant difference in the ASMR of lung cancer and esophageal cancer between Daishan county and the whole nation(lung cancer:28.06/105;esophageal cancer:7.61/105)was observed. Conclusion:Lung cancer,gastric cancer,liver cancer and colorectal cancer were malignant tumors with higher incidence and mortality rates in Daishan county,Zhoushan from 2014 to 2019.Particularly,the ASMR of gastric cancer,liver cancer and colorectal cancer were significantly higher than the national levels,and these malignant tumors should be considered as the major focus of cancer prevention and control.

Background and purpose:Along with the development of diagnosis and treatment technology, the disease free survival and overall survival of breast cancer have been extended. In order to improve the quality of life after mastectomy, more and more breast reconstructions were applied in breast cancer patients. We retrospectively reviewed 118 cases of free abdominal lfaps for breast reconstruction performed in Fudan University Shanghai Cancer Center. Clinical outcomes, reconstructive techniques and experiences are discussed. Methods:From November, 2006 to June, 2013, we used free abdominal lfaps to perform 118 cases of breast reconstruction on 117 female patients after mastectomy. We observed the surgery, complications and safety of this technic. Results:We performed 118 cases of lfaps based on deep inferior epigastric vessels. The average operation time was 7.72 h. The average time of ischemia was 78.74min. The average anastomosis time was 60.83min. The average number of perforators included in the lfap was 3. The internal thoracic vessels were preferred recipient vessels. Ten cases of vessel crisis occurred and 6 of them were venous thrombosis and 4 cases were venous kink. Seven of them were salvaged, and the other 3 failed, the success rate was 97.46%. Postoperative infection rate was 7.00%. Abdominal bulge occurred in 3.50%of patients. None of the patients developed abdominal hernia. The median interval between surgery and the ifrst cycle of adjuvant chemotherapy was 19 days. The median follow-up time was 12 months. One case of distant metastasis, but no local recurrence was observed. Conclusion: Although free abdominal flap breast reconstruction requires complicated microsurgical techniques, and the learning curve does exist, free abdominal lfap breast reconstruction has a high success rate with oncological safety and few complications.