Objective: To observe the effects of Zhichuanxiaoke strong solution on the immunity and irritability of mice. Methods: The carbon clearance index and the content of serum hemolysin of mouse were determined by colorimetry. The survival time of mouse was measured through hypoxia tolerance test under ordinary pressure and the anti fatigue swiming test. Results: Zhichuanxiaoke Strong Solution could obviously increase the carbon clearance index and the content of serum hemolysin, enhance the anti hypoxia ability and prolong the swiming time of mouse. Conclusion: Zhichuanxiaoke Strong Solution can increase physical immunity and the ability of auti irritability.

AIM:To investigate the effects of survivin inhibitor YM155 {4,9-dihydro-1-(2-methoxyethyl)-2-methyl-4,9-dioxo-3-(2-pyrazinylmethyl)-4,9-dihydro-1H-naphtho[2,3-d]imidazolium bromide} on the apoptosis, mito-chondrial membrane potential (Δψm) and cytochrome C (Cyt C) of retinoblastoma Y79 cells, and to analyze the mitochon-drial mechanisms of apoptosis .METHODS:Y79 cells were cultured in vitro and treated with YM155 at concentrations of 0, 0.5, 1, 2, 4 and 8 nmol/L.The cells in control group were treated without YM 155.The proliferation of Y79 cells were measured by CCK-8 assay and bromodeoxyuridine ( BrdU) labeling assay .Y79 cells were randomly divided into 4 groups:control group ( with equal volume of RPMI-1640 nutrient medium ) , positive control group ( 10 nmol/L topotecan ) , low-dose (1 nmol/L) YM155 group and high-dose (2 nmol/L) YM155 group.The effects of YM155 on the apoptosis, the changes of Δψm , the mitochondrial distribution and the protein level of Cyt C in the Y 79 cells were evaluated by flow cytom-etry with Annexin V-FITC/PI staining, JC-1 staining, immunofluorescence analysis and Western blot , respectively.RE-SULTS:Compared with control group , YM155 significantly inhibited the proliferation of Y 79 cells and induced apoptosis (P<0.05).YM155 significantly reduced Δψm of the Y79 cells, promoted Cyt C which released from mitochondria to the cytosol and reduced the protein level of Cyt C in the mitochondria (P<0.05).CONCLUSION:YM155 inhibits Y79 cell proliferation and induces apoptosis , and the possible mechanisms may be involved in the mitochondrium-mediated apoptotic pathway .

Objective To explore the knowledge about HIV/AIDS and related factors associated among farmer workers with private corporations in Guangxi and to inform intervention programs.Methods Questionnaire survey was conducted among workers working with 10 private corporations,and data were analyzed using logistic regression.Results Correct answers to questions about HIV/AIDS transmission route accounted for over 85%,yet much less regarding knowledge about non-transmission routes and prevention methods.The factors associated with knowledge about HIV/AIDS included gender,age,nationality,registered living place and years of schooling.Conclusions HIV/AIDS-related education should be strengthened for farmer workers working for private corporations.

Objective The detection results of intestinal virus nucleic acid related to HFMD( hand,foot and mouth disease)from the anal swabs and throat swabs were compared so as to explore more effective methods for laboratory diagnostic of the disease. Methods Real-time fluorescence quantitative polymerase chain reaction (PCR)was used to detect RNA of enterovirus 71(EV71),coxsackievirus A16(CA16),and other enterovirus (EV)in the paired samples. The virus detection results from the two sample types were compared to find out the miss rate in a single sample. Results The detection rates of EV71,CA16 and Non-EV71/Non-CV16 enterovirus (Non-EV71/Non-CV16 EV)in the anus swabs/ throat swabs were 17.25%/12.18%,4.03%/3.54% and 40.28%/45.32%,respectively. Obviously ,the detection rates of EV71 and CA16 in anal swabs were higher than those in throat swabs ,while the detection rates of Non-EV71Non-CV16 EV was lower than that in throat swabs. The differences were statistically significant(P<0.05). Compared with the detection results of the paired samples ,the miss rate of a virus in a single sample type ranged from 10.33%to 36.27%. Conclusion The detection miss rates of HFMD related intestinal virus nucleic acid in one sample type(anal swabs or throat swabs)are high,while tak-ing the detection in the paired samples can significantly improve the detection rate.

Objective To evaluate the protective effects and mechanism of levocarnitine preconditioning (LCN) on myocardial is-chemia-reperfusion injury in patients undergoing cardiopulmonary bypass .Methods 60 cases of ASA Ⅱ or Ⅲ degree and NYHAⅡ or Ⅲ degree patients who aged 25 ~ 57 years old ,undergoing cardiopulmonary bypass with elective cardiac valve replacement were randomly divided into 2 groups (n = 30 each) :group C (treated with 0 .9% sodium chloride) and group L (treated with LCN) .Group L was infused levocarnitine 50 mg/kg per 1 day at the beginning of 7 days before operation ,group C was given the same amount of 0 .9% sodium chloride .Blood samples were taken from central vein at 5 min after the induction the level of anesthe-sia (T0 ,baseline) ,5 min before aortic cross-clamping (T1) ,30 min after release of the aortic cross-clamp (T2) and at 6 (T3) ,12 (T4) and 24 h (T5) after operation for determination .The level of plasma cardiac troponin I (cTnI) ,creatine kinase-MB (CK-MB) and tumor necrosis factor-α (TNF-α) .Myocardial specimens were obtained from right auricle before aortic cross-clamping and after release of aortic cross-clamp to observe the pathologic changes ,the protein expression of p38 MAPK and phosphorylational-p38 MAPK that analyzed by western blotting .Cardiac index (CI) and left ventricular ejection fraction (LVEF) were measured at 1st day before operation and 7th day after operation by using heart color ultrasonography .Results The levels of cTnI ,CK-MB and TNF-α were significantly lower at all time points in group L than in group C (P< 0 .05) .Myocardial mitochondrion impairment was lighter ,the expression of p38 MAPK and phosphorylational-p38 MAPK were significantly attenuated in group L than in group C (P< 0 .05) .CI and LVEF were significantly higher at 7th day after operation in group L than in group C(P< 0 .05) .Conclusion Le-vocarnitine preconditioning can attenuate myocardial ischemia-reperfusion injury and recover cardiac function in patients undergoing cardiopulmonary bypass ,the mechanism may be related to keep the integrity of the mitochondrial membrane and space structures , inhibit the expression of p38 MAPK and phosphorylational-p38 MAPK and decrease the inflammatory response .

Objective To investigate the expression of c-FLIPL in leukemia and explore its clinical significance. Methods The expression level of c-FLIPL mRNA in bone marrow mononuclear cells was determined by real-time polymerase chain reaction(PCR)in 103 leukemia patients with different types of leukemia,including 54 cases of acute lymphocytic leukemia(ALL)with 37 newly diagnosed,5 relapsed,and 12 complete remis-sion,38 cases of acute myeloid leukemia(AML)with 24 newly diagnosed,6 relapsed,and 8 complete remission,newly diagnosed 2 cases of acute undifferentiated leukemia(AUL),6 cases of chronic myelocytic leukemia(CML),and 3 cases of chronic myelomonocytic leukemia(CM-ML). The immunophenotype of patients were detected by flow cytometry. Results Expression level of c-FLIPL mRNA was higher in newly diag-nosed and relapsed leukemia patients. There was no significant difference between newly diagnosed and relapsed leukemia patients(P>0.05). Ex-pression level of c-FLIPL mRNA of AUL and CML was higher than that in other patients ,but there was no significant difference(P>0.05). Ex-pression level of c-FLIPL mRNA of all newly diagnosed and relapsed leukemia patients was significantly higher than that in control group and com-plete remission group(P<0.05). The expression level of c-FLIPL mRNA was correlated with risk stratification ,white blood cell(WBC),lactate dehydrogenase(LDH),hydroxybutyrate dehydrogenase(HBDH),CD45 and TEL-AML1,but was not associated with age,sex,fibrinogen and chromosome abnormality. Conclusion c-FLIPL mRNA is highly expressed in leukemia patients ,and is closely related with risk stratification , WBC,LDH,HBDH and prognosis.

Objective To explore the effects of inhospital emergency process ~engineering for patients with acute poisoning.Methods Emergency Department of the Affiliated Hospital of Hangzhou Normal University implemented inhospital poisoning emergency process reengineering in March 2016.This implementation optimized original emergency process and applied it in patients with acute poisoning beginning with 6 aspects including refining precheck patients,assessment of poisoning emergency response group,fast gastrolavage,transportation,gastrolavage combined with blood purification group rapid preparation,emergency intensive care unit preparation.We compared the rescue efficiency of patients with acute poisoning before (from March 2015 to February 2016) and after (from March 2016 to February 2017) process reengineering.Results After process reengineering,the time from being admitted to hospital to beginning gastrolavage and the duration of gastrolavage was (8.91 ± 5.29)min and (31.86 ± 8.42)min respectively shorter than those before process reengineering with significant differences (t=3.397,4.028;P ＜ 0.01).After process reengineering,the time from being admitted to hospital to opening blood purification tubes (176.59 ± 88.73)min and from being admitted to hospital to starting blood perfusion (229.35 ± 108.79)min were significantly sooner than those before process reengineering (t=3.600,3.550;P ＜ 0.01).Conclusions The inhospital emergency process reengineering is scientific and convenient.It is propitious to improve rescue efficiency of patients with acute poisoning.

To investigate the effect of hSCGF-alpha on human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs), we obtained hSCGF-alpha using genetic engineering, hSCGF-alpha gene was amplified from hUCMSCs cDNA using two-step PCR and was inserted into pET-28a(+) plasmid vector. Induced by IPTG at 20 degrees Celsius for 24 h, the fusion protein expressed in E. coli BL21 (DE3) was mainly existing in soluble form. The recombinant hSCGF-a was purified using NI-NTA affinity chromatography and the purity was up to 90%. The colony forming test revealed that combined use hSCGF-alpha and rmGM-CSF (recombinant murine GM-colony stimulating factor, rmGM-CSF) had granulocyte/macrophage (GM) promoting effects on murine bone marrow GM progenitor. In addition, the results indicated that hSCGF-alpha and rhGM-CSF had stimulatory effect on hUCMSCs and their synergetic effect was the strongest.
Cell Proliferation ; drug effects ; Cells, Cultured ; Cloning, Molecular ; Drug Synergism ; Escherichia coli ; genetics ; metabolism ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Stem Cell Factor ; biosynthesis ; genetics ; Umbilical Cord ; cytology

Cyanovirin-N (CVN) is an 11 kDa anti-HIV protein originally isolated from extracts of a cyanobacterium, Nostoc ellipsosporum. The protein binds with high affinity to the viral envelope glycoprotein gp120 and irreversibly inactivates diverse HIV strains. A fusion gene consisting of cvn, sumo and 6xHis tag was synthesized by PCR according to the codon bias of Escherichia coli. The fusion protein is expressed in the cytoplasm of E. coli in a soluble form and up to 28% of the total protein. The recombinant CVN was purified to homogeneity by 2 rounds of Ni-NTA affinity chromatography and one round of SUMO protease cleavage. Bioactivity assay demonstrated that SUMO-CVN and CVN bound to gp120 with nanomolar concentration. In addition, CVN showed potent anti-HSV-1 and anti-HIV-1 activities in in vitro cellular assays. Therefore, the 6xHis SUMO fusion expression and purification system provides a better approach for large scale production of CVN for further microbicide development.
Antiviral Agents ; isolation & purification ; metabolism ; pharmacology ; Bacterial Proteins ; biosynthesis ; genetics ; pharmacology ; Carrier Proteins ; biosynthesis ; genetics ; pharmacology ; Escherichia coli ; genetics ; metabolism ; HIV-1 ; drug effects ; Herpesvirus 1, Human ; drug effects ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; pharmacology

OBJECTIVETo evaluate the clinical value of multicolor melting curve analysis(MMCA) for detecting genetic mutations in G6PD deficiency.

METHODSA total of 402 peripheral blood samples(256 males and 146 females) were collected from suspected patients or their relatives at the Prenatal Diagnosis Center of Liuzhou Maternal and Child Health Hospital between March 2012 and May 2012. The samples were screened by G6PD/6PGD quantitative ratio testing. The reliability of the assay was evaluated by multiplex probe melting curve assay(which can detect 16 G6PD mutations) and DNA sequencing through a double blind study.

RESULTSOne hundred seventy cases with G6PD/6PGD ratio < 1.0 and 232 cases with G6PD/6PGD ratio ≥ 1.0 were detected by the enzymological method. DNA sequencing has identified 182 wild type samples, 151 hemizygous mutation samples, 5 female homozygous mutation samples, 54 female heterozygous mutation samples and 10 female double heterozygous mutation samples. Multicolor melting curve analysis has detected 185 wild type samples, 148 hemizygous mutation samples, 5 female homozygous mutation samples, 55 female heterozygous mutation samples and 9 female double heterozygous mutation samples. The specificity and sensitivity of G6PD gene mutation detection by multicolor melting curve analysis were 100%(182/182) and 98.6%(217/220), respectively. The positive predictive value and negative predictive value were 99.5%(216/217) and 98.4%(182/185), respectively, and the Youden's index was 0.986. The concordance rate of the sample detection between the melting curve assay and DNA sequencing was 99.0%(398/402). Twenty-one different genotypes were detected by the multicolor melting curve analysis and 24 different genotypes were detected by DNA sequencing. Four samples containing mutations(c.196T>A or c.406C>T) were not detected by multicolor melting curve analysis, which can be attributed to different technical settings of the two methods.

CONCLUSIONMulticolor melting curve analysis for G6PD gene mutation detection is a simple, rapid, sensitive and specific method, which can be used for clinical diagnosis of G6PD deficiency.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Glucosephosphate Dehydrogenase ; genetics ; Humans ; Infant ; Infant, Newborn ; Male ; Mutation ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA