Repurposed benzydamine targeting CDK2 suppresses the growth of esophageal squamous cell carcinoma / 医学前沿

Esophageal squamous cell carcinoma (ESCC) is one of the leading causes of cancer death worldwide. It is urgent to develop new drugs to improve the prognosis of ESCC patients. Here, we found benzydamine, a locally acting non-steroidal anti-inflammatory drug, had potent cytotoxic effect on ESCC cells. Benzydamine could suppress ESCC proliferation in vivo and in vitro. In terms of mechanism, CDK2 was identified as a target of benzydamine by molecular docking, pull-down assay and in vitro kinase assay. Specifically, benzydamine inhibited the growth of ESCC cells by inhibiting CDK2 activity and affecting downstream phosphorylation of MCM2, c-Myc and Rb, resulting in cell cycle arrest. Our study illustrates that benzydamine inhibits the growth of ESCC cells by downregulating the CDK2 pathway.

Specific expression of CPS-II in hyperammonemia-injured liver cells / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the CPS-II mechanism underlying the pathological process of elevated blood ammonia leading to liver injury.</p><p><b>METHODS</b>An in vitro hyperammonemia hepatocyte cell model was constructed by exposure to various concentrations of NH4Cl. The subsequent changes to cellular morphology were observed by microscopy. to cell apoptosis were determined by flow cytometry, and to mRNA and protein expression of CPS-II were examined by real-time PCR and western blotting, respectively.</p><p><b>RESULTS</b>Exposure to NH₄Cl led to dose-dependent morphological damage, apoptosis and necrosis of the hepatocytes. The apoptosis rate was significantly higher for the high-dose group than for the control (no exposure) group (24.7% ± 2.39% vs. 4.1% ± 0.78%, q =8.06, P less than 0.05). Expression of the CPS-II mRNA was significantly elevated in response to NH₄Cl exposure (vs. the control group; F=191.881, P < 0.05).The CPS-II mRNA expression level increased with increasing NH₄Cl concentration (grey values: 1.040 ± 0.045, 1.641 ± 0.123, 2.285 ± 0.167 and 3.347 ± 0.124, respectively). The CPS-II protein expression level was also significantly enhanced in response to the NH₄Cl exposures (CPS-II protein and internal GAPDH grey value ratios: 0.099 ± 0.0130, 0.143 ± 0.025, 0.161 ± 0.036 and 0.223 ± 0.042, respectively; t=3.825, 3.968 and 6.908, P less than 0.05).</p><p><b>CONCLUSION</b>CPS-II mRNA and protein expression levels become elevated with increase in the NH₄Cl concentrations, suggesting that in addition to the urea cycle, CPS-II may play an important role in the ammonia metabolism under the condition of hyperammonemia.</p>

Expression of reconstructed BCR-ABL-pIRES-SEA plasmids in the skeletal muscles of BALB/c mice / 生物医学工程学杂志

This paper is aimed to investigate the transcription and expression of BCR-ABL-pIRES-SEA fusion gene vaccines in vivo in mice. The reconstructed plasmids (BCR-ABL-pIRES-SEA) which were developed previously in our laboratory were injected into the skeletal muscles of BALB/c mice at 14d intervals for three cycles. The transcription and expression of BCR-ABL and staphylococcal enterotoxin A (SEA) in injection site were detected using RT-PCR and immunohistological methods. The BCR-ABL/SEA mRNA and protein could be identified in the injection site of BCR-ABL-pIRES-SEA vaccinated mice. The reconstructed BCR-ABL-pIRES-SEA plasmids can effectively express gene production in the skeletal muscles of mice and have the common features of DNA vaccine.

Detection of apolipoprotein B-100 gene mutation in patients of primary hypercholesterolemia families / 中国实用内科杂志

Objective To detect genetic mutation of apolipoprotein B-100(apoB-100)in patients with primary hypercholesterolemia.Methods One special segment of apoB-100 gene from nucleotide 10549 to 10895 was amplified by polymerase chain reaction(PCR).The PCR products were denatured and hybridized with specific aligonucleotide labeled with digoxigenin,and were analyzed by single strand conformation polymorphism(SSCP)to detect the apoB-100 gene mutation 3531CGC→CGT or other mutations.Results Overall 41 members of 11 primary hypercholesterolemia families were detected,but the above genetic mutation was not detected.Conclusion This genetic mutation is unlikely to exist or of significantly low incidence in Chinese population,and might not be the main cause of primary hypercholesterolemia in the 11 primary hypercholesterolemia families.

Detection of Human Herpesvirus-6 DNA in Peripheral Blood Cells of Patients with Myelogenous Leukemia by Nested PCR / 中国实验血液学杂志

Human herpesvirus-6 (HHV-6) is largly disseminated in human population, which may relate to many diseases. Due to different methods previously used, the results of HHV-6 DNA detections in patients with leukemia are not identical. With a control-study, HHV-6 DNA in the blood cells from 38 cases, including 30 cases with acute myeloid leukemia and 8 cases with chronic myelogenous leukemia was detected by nested PCR. The results showed that HHV-6 DNA positive rate (63%) in patients with AML was lower than that in controls, whereas no significant difference between patients with CML (75%) and controls (92%) was shown. The clinical importance of HHV-6 infections in myelogenous leukemia should continue to be investigated later.

Changes in TCR V? subfamily nave T cells in peripheral blood of patients with multiple myeloma / 中国病理生理杂志

AIM:To detect the existence of signal joint T-cell receptor excision DNA circles(sjTRECs)of 23 TCR V? subfamilies in mononuclear cells of patients with multiple myeloma(MM),and to evaluate the recent thymic emigrants of corresponding V? subfamily nave T cells in MM patients.METHODS:23 TCR V? subfamily sjTRECs were amplified in genomic DNA from 5?104 PBMCs of 12 cases in MM patients by using semi-nest PCR.10 normal individuals served as controls.RESULTS:The number of detectable V? subfamily sjTRECs was 5.00?2.45 from MM patients,as compared with 9.60?5.48 from normal individuals,the difference was significant(P

Effect of shenmai injection on myocardial eNOS mRAN expression in myocardial ischemic rats / 中国病理生理杂志

AIM: To study the effect of shenmai injection (SMI), a Chinese medicine, on nitric oxide and the expression of endothelial nitric oxide synthase (eNOS) mRNA in myocardium of rats with experimental myocardial ischemia. METHODS: Rats were randomly divided into four groups: control, model (ischemia), SMI 1 and SMI 2 group. A rat model of acute myocardial ischemia was established by isoprenaline treatment and the lift of ST segment in ECG was used as the index of myocardial ischemia. The nitric oxide (NO) contents in serum and myocardium and the expression of eNOS mRNA in myocardium were measured. RESULTS: Compared with control group, ST segment of ECG was significantly elevated 20, 30, 40 min after myocardial ischemia in model group, the lift peak of ST segment occurred 20 min after myocardial ischemia, the concentration of NO in the serum and myocardium and the expression of eNOS mRNA in myocardium were significantly lowered in model group. Compared with the model group, in SMI 1 group and SMI 2 group, the concentration of NO in the serum and myocardium and the expression of eNOS mRNA in myocardium were significantly increased, the lift of ST segment were significantly reduced 20, 30, 40 min after myocardial ischemia. Compared with SMI 1 group, the concentration of NO in the serum and myocardium and the expression of eNOS mRNA in myocardium and the lift of ST segment were not statistically different in SMI 2 group. CONCLUSION: Shenmai injection can increase the expression of eNOS mRNA in myocardium and the content of NO, and protect against myocardial ischemia in rats.

Construction of human neuronal nitric oxide synthase expression system in Escherichia coli / 中国病理生理杂志

AIM: To construct a high-level expression system of recombinant human neuronal nitric oxide synthase (hnNOS) full-length enzyme in Escherichia coli. METHODS: The coding sequence of hnNOS full-length was firstly amplified by PCR, and then ligated into the expression vector pCWori+. The recombinant plasmid was transformed into Escherichia coli BL21 for high-level expression. After having been checked with Western blot, the enzyme was used for large-scale culture and purification. Finally, the property of the enzyme was determined by spectrophotometric method. RESULTS: The constructed expression system could give a yielding of 3 mg/L initial culture. CONCLUSION: The expression system constructed is fully sufficient to express the active human neuronal nitric oxide synthase.