Anthrax is a malignant infectious disease caused by Bacillus anthracis spores,after entering the host Bacillus an-thracis produces and releases anthrax toxin,which is the main cause leading to death of the host. The anthrax toxin is composed of two enzymatically active components:lethal factor(LF)and edema factor(EF),and one shared receptor binding and translocation com-ponent:protective antigen(PA). PA combined with LF is called lethal toxin(LeTx),while PA combined with EF called edema toxin (EdTx). Currently,the main drugs for treating anthrax are antibiotics,but antibiotics can only kill part of anthrax spores and bacte-ria,and cannot inhibit the activity of anthrax toxin. So it is necessary to develop novel drugs for inhibiting anthrax toxin. This review summarizes the evolution of small-molecule inhibitors of anthrax toxin respectively targeting PA,LF and EF.

Objective To construct a lentiviral vector carrying CRKL gene RNA interfering( RNAi ).Methods The CRKL RNAi was selected and subcloned into the lentiviral vector,pGCL-GFP(including U6 promotor and green fluorescent protein),generating the lentiviral vector LV-shCRKL.The corrected CRKL was confirmed by endoenzyme digestion ,sequencing.Recombinant lentiviruses were produced by 293T cells following the eo-transfection of LV-shCRKL,with the packaging plasmids pHelper1.0 and pHelper2.0.The virus titer was detected by GFP expressions in 293T cells.Results Plasmid LV-shCRKL carried the correct sequence.The recombinant lentiviruse LV-shCRKL could be produced by co-transfection of LV-shCRKL to 293T cells.Conclusion The recombinant lentiviruse vector LV-shCRKL is constructed successful.

Objective:To clone the full-length cDNA of human ubiquitious mitochondrial creatine kinase(uMtCK) and express its protien in E. coli. The protein was used to immunize rabbit to prepare anti-uMTCR polyclonal antibody which was then used to detect the expression of uMtck in gastrointestinal carcinoma. Methods: A 1 062 bp segement was amplified from human HaLa cell by RT-PCR,which was then inserted into pMD18-T plasmid and proved to be the coding sequence of uMtCK by endonuclease digestion and sequencing. The coding sequence of uMtCK was inserted into pQE30 plasmid and recombinant plasmid pQE30-uMtCK was transformed into E. coli. The expressed protein was purified with a His. Bind column and was analyzed with SDS-PAGE and Westen blot and the activity of uMtCK was tested. An antiserum against uMtCK was perpetrated by immunizing rabbit with purified uMtCK. The expression of uMtCK in 59 patients with gastric and colonic cancer was detected by immunohistochemistry. Results:The coding sequence of uMtCK was successfully obtained by RT-PCR and the product was successfully inserted into pQE30 to construct a recombinant vector pQE30-uMtCK. The expression of uMtCK was effective and soluble, A good specific antiserum against uMtCK was proved by Western blot. The expression of uMtCK of gastric and colonic cancer was 76. 5%. Conclusion:The coding sequence of uMtCK is coined and expressed in E.coli. uMtCK is a prospective marker for the diagnosis of gastric and colonic cancer.

Objective To investigate the effects of rhubarb extracts, i.e. rhein and emodin, on the neuronal hyperexcitability and synaptic transmission, and to further reveal the mechanism of the secondary brain damage. Methods The fluid percussion injury (FPI) rat model and extracellular recording method were used. The evoked field potentials by stimulating Schaffer collaterals were collected from the ipsilateral (impact side) and the contralateral hippocampal CA1 areas of rat in vitro. And the field potentials, including the field excitatory postsynaptic potential and the population spike, were analyzed. Results After the impact was performed on the rat parietal cortex, the evoked field potentials in the ipsilateral hippocampus CA1 area were enhanced obviously. Rhubarb extracts reduced the slope of the field excitatory postsynaptic potential and the number of the population spike significantly while rhein and emodin increased the latency of the population spike obviously. Conclusion Rhubarb extracts, i.e. rhein and emodin, can depress the neuronal hyperexcitability, which suggests that rhein and emodin play an important role in protecting the central nervous system from neuronal damage after traumatic brain injury. FPI produces hyperexcitability of hippocampal CA1 neurons, probably by enhancing excitatory synaptic transmission.

OBJECTIVE To explore the toxic effects of zinc oxide nanoparticls (ZnO-NPs) on cardiac development of zebrafish embryos and rat myocardial cell lines (H9c2),as well as potential molecular mechanisms.METHODS ZnO-NPs were characterized.Zebrafish embryos were exposed to different doses of ZnO-NPs (0,0.5,2.5,5.0 and 10.0 mg· L-1) for 24 to 96 h at 4 h post fertilization (4 hpf).The embryo mortality was observed.The expressions of notochord homeobox (noto),T-box 6 (tbx16),T,brachyury homolog a(ta),and tbx6 which were related to cardiac mesoderm were investigated using real-time PCR at 17 hpf.The heart rate and number of cardiomyocytes of embryos [[Tg (cmlc2:nucdsRed)] exposed to 0,2.5 and 5 mg· L-1 ZnO-NPs were detected at 72 hpf.Rat myocardial cell lines (H9c2) were treated with ZnO-NPs (0.1,0.5,1.0,5.0 and 10.0 mg· L-1) for 24 h.Cell viability was measured with Alamar Blue method.Mitochondrial ultrastructure was observed by transmission electron microscopy.Cellular ATP was detected using chemiluminescence,and oxygen consumption rate (OCR) was examined with Seahorse instrument.RESULTS The particle size of ZnO-NPs was (331 ±3)nm.The ZnO-NPs LC50 of zebrafish embryos at 48 hpf was 21.81 mg· L-1.The mRNA expressions of noto,ta and tbx6 were reduced after ZnO-NPs 2.5 mg· L-1 treatment at 17 hpf.The heart rate of 72 hpf zebrafish was 153 min-1 in the ZnO-NPs 5 mg· L-1 group,12.6％ lower than that in the cell vehicle group (P＜0.01),and the number of cardiomyocytes decreased by 15.5％ (P＜0.01) compared with the cell vehicle group.Reduced cell viability and mitochondrial vacuolation were observed in H9c2 after ZnO-NPs 0.5 mg· L-1 exposure.Compared with the cell vehicle group,the cell ATP decreased by 25.7％ (P＜0.05),and OCR decreased by 27.2％ (P＜0.01).CONCLUSION Low-dose ZnO-NPs exposure has effect on the cardiac development of zebrafish,mainly due to reduced heart rate and decreased number of cardiomyocytes.These changes may be related to the decreased expressions of cardiac development-related genes and the impairment of mitochondrial structure and function.

Objective To establish the immunoprecipitation method to detect the activity of ?-glutamyltransferase (GGT) complexed to low-density-lipoprotein and very-low-density-lipoprotein in serum (LDL-LDL-GGT) and to observe the diagnostic efficiency in discriminating liver malignancies from other hepatopathy diseases.Methods Using apo B antibody as a precipitant to precipitate the parts of GGT which connected to LDL and VLDL. 65 cases of liver malignancies, 53 cases of cirrhosis, 32 of chronic hepatitis and 75 of health people were analysed. Results Apo B antibody can precipitate LDL and VLDL thoroughly with various serum samples. The withinrun CV are 3 6%～8 2% and the betweenrun CV are 5 5%～9 8%. The linearity range is 0～587 U/L. With the cut off value of 10 U/L, the sensitivity for liver malignancies is 86 2%, the specificities for discriminating liver malignancies from cirrhosis and chronic hepatitis is 67 9% and 81 3% respectively. Conclusion It has important significance for the diagnosis of liver malignancies with the immunoprecipitation method for determination of serum LDL-VLDL-GGT activity.

Objective To explore the application effect of interactive health education in insulin injections of aged patients with type 2 diabetes in the incipient stage of diagnosis and treatment. Methods 100 cases of aged patients with type 2 diabetes were hospitalized into endocrinology department from January 2014 to December 2014, the pen insulin injections were conducted for them at the beginning. They were equally divided into experimental group and control group through random number method. And, the general education guidance was conducted in control group. Meanwhile, the diabetes health education team was established in observation group on the basis of control group. Then, it was educated through the interactive lectures, interviews, teaching, repeated teaching, psychological intervention and individual assessment. Before and after education, the KAP survey of standard insulin injections were used for the patients in two groups to evaluate their degree of proficiency in the knowledge and skills about insulin injections, and the incidence of glucose metabolism level and low blood sugar. Results After intervention, the insulin injection skill in observation group was better than that of in the control group, (64.6%,31/48 vs. 24.5%, 12/149,χ2=16.441, P<0.01), while the level of glycosylated hemoglobin was reduced from (7.95±0.65)%to (6.58±0.62)%(t=7.85, P<0.01). Conclusions In the incipient stage of diagnosis and treatment, the ability of aged patients with type 2 diabetes in learning new knowledge and technology was improved through interactive health education mode. Besides, the insulin injections were standardized. Also, the glucose metabolism was improved, and the incidence of hypoglycemia was significantly reduced to ensure the patients′safety when the insulin was injected outside hospital, which was worth being clinically applied and promoted.

Objective To discuss the application effect of peer support education in improving theself-management ability of old patients with diabetes. Methods A total of 160 cases of old patients with type 2 diabetes who were treated and hospitalized from June 2014 to August 2015 were selected, then equally divided into observation group and control group through using random number method.Finally, 76 cases were studiedin observation group, while 75 cases in control group. The health education guidance of conventional diabetes had been used in two groups for six months, and the peer support education was also conducted for the patients in observation group on the basis of conventional education. The self-management ability, insulin standard injection knowledge-belief and attitude-behavior/practice (KAP) and glycosylated hemoglobin (HbA1c) value of old patients with diabetes were observed before intervention, 3 months and 6 months later, respectively. Also,the hypoglycemia frequency in two groups was compared during intervention. Results After three months of intervention, the score of diet, exercise management, glucose monitoring, foot care, pharmaceutical administration and smoke for self-management ability in observation group was 4.63 ± 0.92, 4.18 ± 0.97, 2.86 ± 0.96, 2.35 ± 0.59, 4.01 ± 1.43 and 0.62 ± 0.29, respectively, while that of these indicators in control group was 3.69 ± 0.83, 3.52 ± 0.67, 1.75 ± 0.67, 1.63 ± 0.72, 3.05 ± 1.39 and 0.59 ± 0.38. Except smoke (P > 0.05), the differences between two groups were statistically significant (t=2.987-5.965, P < 0.05). After six months of intervention, the score of diet, exercise management, glucose monitoring, foot care, pharmaceutical administration and smoke for self-management ability in observation group was 5.86±0.93, 5.37±0.61, 3.97±0.94, 5.06±0.92, 5.81±0.73 and 0.89 ± 0.31, respectively, while that of these indicators in control group was 3.68 ± 0.96, 3.65 ± 0.95, 1.86 ± 0.88, 2.65±0.88, 3.21±1.13 and 0.81±0.39. The difference was statistically significant (t=9.336-15.633, P<0.01), except smoke (P>0.05). Then, the insulin was normally injected in observation group after six months of intervention, the score of knowledge, attitude and behavior was 5.63 ± 1.87, 43.17 ± 6.03 and 16.57 ± 1.59, while that in control group was 3.76 ± 0.83, 23.71 ± 0.59 and 10.16 ± 0.95. The differences between two groups were statistically significant (t= 8.569, 15.623, 6.751, P<0.01). Also, the level of glycosylated hemoglobin was decreased from (8.17 ± 0.32) % to (7.16 ± 0.41) %, and the difference was statistically significant (t=-3.560, P=0.000). Conclusions Peer support education couldenhance the self-management consciousness of old patients with diabetes, standardize the insulin injection, improve the ability of self-management and improve glucose metabolism index so that it is worthy of being clinically popularized and applied.

Phospholipase C (PLC) is a key enzyme in phosphatidyl inositol turnover during signal transduction. The 12 mammalian PLC isozymes identified to date can be divided into five subtypes, beta-type, gamma-type, delta-type, epsilon-type and zeta-type, with extensive difference in structure, regulatory mechanism and tissue distribution. PLC plays important roles not only in sperm acrosome reaction but also in egg activation. The present studies are reviewed on the structure, regulation and function of PLC, especially its function in male reproduction, including triggering Ca2+ oscillations in eggs to activate the eggs and helping embryo development. And the prospect of the clinical application of PLC is discussed.
Acrosome Reaction ; Animals ; Calcium Signaling ; Humans ; Reproduction ; physiology ; Signal Transduction ; Type C Phospholipases ; chemistry ; physiology

AIM To study the influence of ginkgo biloba exocarp polysaccharides(GBEP) on p53 gene expression and telomerase activity of human gastriccancer SGC 7901 cells in vitro . METHODS The p53 gene expression level of human gastriccancer SGC 7901 cells were measured by immunohistochemistry ABC. The telomerase activity of human gastriccancer SGC 7901 cells were measured by TRAP ELISA. RESULTS GBEP(80～160 mg?L -1 ,48 h) inhibited the expression of mutated type p53 genes and telomerase activity of human gastriccancer SGC 7901 cells. CONCLUSION The action mechanism of GBEP that inhibited the proliferation of SGC 7901 cells may relate to its down regulation effect on the expression of p53 genes and telomerase activity of human gastriccancer SGC 7901 cells.