The aim of this study is to construct specific shRNA expressing plasmids, and to observe their effects on H9c2 cardiomyocytes injury induced by hypoxia/reoxygenation (H/R). RIPK1 and RIPK3 are the key kinases mediating the process of necroptosis. Using recombinant DNA technology, we inserted the synthetic shRNA into pSUPER vector to construct RIPK1-shRNA or RIPK3-shRNA plasmid respectively. We transfected H9c2 cardiomyocytes with the two shRNA plasmids respectively, before we treated them with H/R stimulation. Then, we measured the relevant genes and proteins by real-time PCR and Western blot. Meanwhile,we detected the markers of necroptosis and cardiomyocytes injury. The results showed that inhibition of ripk1 or ripk3 gene expression by its specific shRNA might protect the cardiomyocytes injury induced by H/R stimulation.
Animals ; Apoptosis ; Cell Hypoxia ; Cell Line ; Gene Expression ; Myocytes, Cardiac ; pathology ; Protein-Serine-Threonine Kinases ; genetics ; metabolism ; RNA, Small Interfering ; Rats ; Real-Time Polymerase Chain Reaction ; Receptor-Interacting Protein Serine-Threonine Kinases ; genetics ; metabolism ; Transfection

Microfluidic chip is a novel technology platform, in which microchannels are fabricated in different materials. The ability to precisely control the microflows makes it possible to mimic the microenvironment of cells in physiological or pathological states, which provides many distinct advantages for cell research. In this paper are reviewed the design and fabrication of microfluidic chip, the application of microfluidic chip in cell culture and cell researches; the enormous advantages of microfluidic chips in precise experimental control of the cellular microenvironment are introduced.
Cell Adhesion ; Cell Culture Techniques ; Cell Movement ; Cells, Cultured ; Cellular Microenvironment ; Humans ; Microfluidic Analytical Techniques ; methods

Objective To explore the clinical efficacy of laparoscopic left hemihepatectomy for the treatment of intrahepatic bile duct stones.Methods The clinical data of 30 patients with left intrahepatic bile duct stones who were admitted to the Second Affiliated Hospital of Nancbang University from June 2013 to June 2014 were retrospectively analyzed.All the patients underwent laparoscopic left hemihepatectomy by the Glisson intra-and extra-pedicles vascular inflow occlusion techniques together with the removal of choledocholithiasis and right bile duct stones,and T tube placement or laparoscopic primary suture of common bile duct were selected according to the condition of bile duct.All the 30 patients were readmitted to hospital and detected by color Doppler ultrasound (CDUS),computed tomography (CT) and T tube cholangiography at postoperative month 1,and then received CDUS reexamination every 3 months.CT and MRI reexaminations were applied to patients with complication of residual stones if necessary.All the patients were followed up till July 2014.Results All the 30 patients were treated by laparoscopic hepatectomy with left hemihepatic vascular inflow occlusion,including 5 with conversion to open surgery and 25 with successful operation.The Glisson extra-and intra-pedicel vascular inflow occlusion techniques were used in 11 and 14 patients,respectively.The operation time and volume of blood loss were (158 ± 85) minutes and (405 ± 215) mL.Two patients received intraoperative blood transfusion.There were no residual stones in the 8 patients with choledocholithiasis by intraoperative choledochoscope,and primary suture of bile duct and T tube placement were done in 5 and 3 patients,respectively.No patients died.After operation,there were 2 patients with bile leakage and 1 with pleural effusion,and they were cured though drainage.One patient with subphrenic effusion was cured by B ultrasound-guided puncture and drainage.One patient had bleeding with the volume of blood loss of 500 mL,and was cured by conservative treatment.The duration of hospital stay in all the patients was (8.5 ± 2.3)days.No bile leakage and abdomen infection were detected by outpatient examination.The time of followup was 1-12 months,without recurrence of stones.Conclusion Laparoscopic left hemihepatectomy for the treatment of left intrabepatic bile duct stones is safe and feasible with satisfactory outcome.

Objective To investigate the application of statins in secondary prevention of ischemic stroke and transient ischemic attacks (TIA) in different risk groups,and to identify the factors influencing the compliance of statins. Methods Data were prospectively collected on consecutively encountered ischemic stroke or TIA patients admitted to the First Affiliated Hospital of Zhengzhou University from April 2009 to January 2010.All clinical characteristics and possible factors influencing the compliance of statins were collected; the application of statins was investigated at 3-month follow-up.The multivariate Logistic regression analysis was used for the analysis of influence factors of the compliance of statins.Results All 369 patients were collected,52.8％ of cases were prescribed statins for therapy during hospitalization.The application rate of statins in accordance with guidelines in high-risk group,extremely high-risk Ⅱ group and extremely high-risk Ⅰ group was 25.0％ (16/64),44.1％ (30/68) and 71.4％ (135/189),respectively. Logistic regression analysis showed that the statins application during hospitalization was associated with diabetes history ( P =0.032,OR =1.789,95％ CI 1.052-3.043 ) and the presence of carotid vulnerable plaques(P =0.000,OR =5.308,95％ CI 3.340-8.434).The general application rate of statins was 22.3％ (81/363),which was significantly lower than that during hospitalization. The application rate of statins in accordance with guidelines in high-risk group,extremely high-risk Ⅱ group and extremely high-risk Ⅰ group was 9.7％ ( 6/62 ),25.8％ (17/66) and 29.4％ (55/187) respectively.Logistic regression analysis showed that good compliance of statins was associated with discharge instructions on statins application ( P =0.000,OR =34.852,95％ CI 14.673-175.452 ). Conclusions The compliance of statins in secondary prevention of ischemic stroke and TIA is poor,and there is still a large gap between clinical practice and guidelines; Discharge instructions on statins application increase the compliance of statins.

Objective To investigate the predictive value of the modified National Institutes of Health Stroke Scale ( mNIHSS),measured within 9 days after stroke,for the outcome in terms of modified Rankin Scale (mRS) at 6 months after stroke.Methods AII 161 patients with acute anterior circulation ischemic stroke were recruited consecutively from July 2010 until November 2010.The mNIHSS score was assessed within 3 days,4 to 6 days and 7 to 9 days after stroke,and the mRS score at 6 months after stroke was assessed by a neurology resident.Spearman rank correlation and ROC curve were used for statistic analysis.Results One hundred and sixty-one patients were assessed within 3 days,at 4 to 6 days and 7 to 9 days after stroke.Significant Spearman rank correlation coefficients were found between mRS at 6 months and mNIHSS scores within 3 days (rs =0.592,P＜0.01 ),4 to 6 days (rs =0.597,P＜0.01 ) and 7 to 9days (rs =0.595,P＜0.01).At 6 months after stroke,the AUC ranged from 0.789 (95％ CI 0.708-0.856) for measurements within 3 days to 0.792 (95％ CI 0.712-0.857 ) and 0.799 ( 95％ CI 0.721-0.864) for 4 to 6 days and 7 to 9 days,respectively. There was no significant difference for the AUCbetween them.The optimal cut-off values for the prediction of prognosis at 6 months were 8,7 and 6 points of mNIHSS score within 3 days,4 to 6 days and 7 to 9 days,respectively.The negative predictive value gradually increased from 0.847 for assessment within 3 days to 0.867 ( 95％ CI 0.578-0.765 ) for 7 to 9 days,whereas positive predictive value declined from 0.601 for assessment within 3 days to 0.533 for 7 to 9 days.The overall accuracy of predtictions decreased from 0.753 for assessment within 3 days to 0.709 for 7 to 9 days.Conclusions When measured within 9 days,the mNIHSS has a good predictive value for final outcome in terms of mRS at 6 months after stroke.However,the optimal cut-off values for the prediction of prognosis at 6 months are different when early mNIHSS scores were assessed at different periods.

Objective To optimize the formulation of the sunscreen used in military training.Methods The different formulations of sunscreen were prepared according to the orthogonal design,and then applied on 3M adhesive tape posted on the color dish with a certain amount;the absorbance in 280 nm-320 nm and 320 nm-400 nm of them were measured and the arithmetic mean (A) was concluded.By comparing A,the protective effect and the main factors affecting preventive bask were determined.The preventive bask test of the optimum formula was carried out in human trials using the method of multi point increasing ultraviolet irradiation.Results OMC,TinosorbTM M,rutin,pearl powder and baicalin all had significant effects in preventing UVA.OMC had a significant effect in preventing UVB.The formula 1 (containing the five materials mentioned above) had the best effects in preventing both UVA (SPF 18) and UVB (PA+).Conclusions The formula 1 has the best effects in preventing both UVA and UVB.

Objective To study the expressions of osteopontin (OPN),caspase-3 and mt-P53 proteins, and their relationship in gliomas. Methods Seventy gliomas specimens of patients (glioma group) were selected, and 10 samples of non-glioma brain tissue were used as control group. The SP method was used to detect the positive rates of protein expressions of OPN, caspase-3 and mt-P53 between two groups. The relationship between protein expressions of OPN, caspase-3 and mt-P53 in gliomas and grade of gliomas were detected by Western blot assay. Spearman rank correlation was compared between the positive expression of OPN, caspase-3 and rate mt-P53. Results The positive expression rates of OPN and mt-P53 were significantly higher in glioma group (64.29%and 60%) than those of control group (no positive expression), but the positive expression rate of caspase-3 was significantly lower than that of control group (47.14%vs. 90%, P<0.05). There were no significant differences in OPN, caspase-3 and mt-P53 expressions between different gender, age, tumor size and tumor position (P>0.05). The higher the WHO classification, the higher the positive expression rates of OPN and mt-P53 (P<0.001), and the lower the positive expression rate of caspase-3 (P<0.001). With the increased level of glioma grade, OPN and mt-P53 protein levels were increased, but caspase-3 protein expression level was decreased. There was a negatively correlation between OPN and the positive expression of caspase-3, but there was a positive correlation between OPN and the expression of mt-P53 (rs=-0.720 and 0.722, P<0.05). There was a negative correlation between caspase-3 and mt-P53 expressions (rs=-0.556, P<0.05). Conclusion The higher the WHO classification, the higher the positive expression rates of OPN and mt-P53, while the lower the positive expression rate of caspase-3. The study reveals that OPN, caspase-3 and mt-P53 expressions are associated with the occurrence and the progress of gliomas. The combined detection of them can contribute to the judgment of biological behavior of gliomas.

AIM:ToinvestigatetheeffectofP21-activatedkinase1(PAK1)andlymphoidenhancer-binding factor 1(LEF1) on the proliferation of esophagus cancer cells .METHODS:Real-time PCR was applied to detect the mR-NA expression of PAK1 and LEF1 in the esophagus cancer tissues .MTT assay were used to measure the proliferation of hu-man esophagus cancer cell line KYSE transfected with PAK 1 and LEF1.RESULTS: The mRNA expression of PAK1 in the esophagus cancer tissues was lower than that in control group (P<0.05).The mRNA expression of LEF1 and tran-scription factor 4 (TCF4) in the esophagus cancer tissues was higher than that in control group (P<0.05).The prolifera-tion of KYSE cells with over-expression of PAK1 and LEF1 was higher than that in control group .No significant change of apoptosis between the KYSE cells with over-expression of PAK1 and LEF1 and control group was observed .CONCLU-SION:The expression of PAK1 decreases and the expression of LEF 1 increases in esophagus cancer tissues .LEF1 domi-nantly regulates the proliferation of esophagus cancer cells .

Objective:This study aimed to investigate the possible mechanism of 32P colloid induced apoptosis of craniopharyngi-oma (CP) cells in vitro and the relationship between dose effect and time effect. Methods:This study established a primary cell culture of CP limited subculture cell line. Methyl thiazolyl tetrazolium (MTT) assay was performed to plot the cell survival curve after the CP cells were treated with 32P colloid at different concentrations and time. Apoptotic rate was detected by flow cytometry(FCM). Apoptosis related DNA was investigated by TUNEL fluorescent staining. The morphological characteristics of apoptotic cells were determined by Hoechst33342 fluorescence staining. The ultrastructure of apoptotic cells was investigated by transmission electron microscopy (TEM). Results:Hoechst33342 fluorescence staining, TUNEL fluorescence staining, and TEM revealed that 32P colloid induced the apoptosis of CP cells. 32P colloid reduced the survival rate and increased the apoptotic rate of CP cells as concentration (0 MBq/mL to 14.80 MBq/mL) and time (1 d to 14 d) were increased. Conclusion: 32P colloid could effectively inhibit the growth of CP cells and induce apoptosis in vitro. High concentrations and prolonged time could induce a remarkable effect.

Objective:To identify anti-dipeptidyl-peptidase-like protein 6 (DPPX) antibody in patients with encephalitis of unknown etiology and describe the clinical features of anti-DPPX antibody-associated encephalitis in Chinese patients.Methods:For patients registered in the Peking Union Medical College Hospital Encephalitis and Paraneoplastic Syndrome Registration Project from 2016 to 2019 with negative findings in autoimmune encephalitis routine antibody profile and paraneoplastic antibody profile, but with positive tissue-based assay (TBA) results, further tests for rare antibodies, including cell-based assay (CBA) of anti-DPPX antibody, were performed. Patients positive for anti-DPPX antibody were enrolled and the clinical data were collected.Results:Two patients with anti-DPPX antibody-associated encephalitis were found from 2016 to 2019 among about 15 000 patients. Both were females, aged 46 and 75 years. One patient had diarrhea, cachexia, cognitive dysfunction, agitation, myoclonus, tremor, and seizures. The other had cognitive impairment, restlessness, memory loss, disorientation, and sleep disturbance. The second patient had medical history of systemic lupus erythematosus and secondary Sj?gren′s syndrome.Conclusions:TBA should be combined with CBA in identification of anti-DPPX antibody to confirm the diagnosis. Anti-DPPX antibody-associated encephalitis has clinical manifestations of encephalopathy with diarrhea and cachexia, and can coexist with systemic lupus erythematosus.