Constructing prokaryotic expression vector pKY-RMBAY by gene recombination and research its optimizing productive conditions.By PCR technology synthesizing the gene of the RMBAY with preference codon of E.coli and the RMBAY gene was inserted into high efficiency expression vector pKYB-MCS.Expressed fusion proteins in E.coli ER2566 were purified with Chitin-Beads column.Fusion proteins binding on Chitin-Beads was cut on N-terminus of intein due to the induction of ?-mercaptoethanol and the target peptide RMBAY was released.The RMBAY was identified by mass spectrum.Experiment results showed RMBAY can be high efficiently expressed in E.coli ER2566,with optimizing productive conditions the yield of the RMBAY may be 6.7mg/L fermentation product and its purity is greater than 98%.The molecular weight of RMBAY is 3.887 kDa by mass spectrum and that accords with its theory value.

Objective: To compare the efficacy and outcomes of gamma knife stereotactic radiosurgery and microsurgery in small- and medium-sized acoustic neuroma. Methods: The medical records of 87 patients with unilateral acoustic neuroma less than 3 cm were retrospectively collected. These 87 patients were divided into two groups: gamma knife stereotactic radiosurgery (GSR) group (n = 42) and microsurgery (MS) group (n = 45). The local control rate of tumor, preservation rate of cranial nerve function, hospitalization cost in first admission, length of stay in hospital, and the complications were campared between the two groups. Results: The difference in local control rate of tumor was not significant (GRS group vs MS group: 88.1% vs 97.8%; P = 0.102). In respect of preservation of facial nerve function and hearing, GRS was more effective than MS (87.2% vs 66.7%, P = 0.029; 68.2% vs 30.0%, P = 0.013). The rate of complications in the MS group was much higher than that in the GRS group (22.2% vs 7.1%). As compared with the GRS group, the hospitalization cost in first admission of the MS group was higher and the length of stay in hospital was prolonged. Conclusion: GSR has much more advantages in treatment of small- and medium-sized acoustic neuroma as compared with MS. Copyright © 2014 by TUMOR.

OBJECTIVETo investigate the relationship between C7673T polymorphism of apolipoprotein B (apoB) and cerebral hemorrhage with family history (CHFH) in Chinese Han in Changsha, Hunan province.

METHODSFifteen families of CHFH and 93 sporadic cerebral hemorrhage patients and 100 normal controls were collected. The C7673T polymorphism of apoB was analyzed by PCR-restriction fragment length polymorphism and direct DNA sequencing. The triglyceride(TG), total cholesterol(TC), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol(LDL-C) levels were examined by oxidase method. The serum level of lipoprotein (a) was determined by immune method.

RESULTS(1)The allele T frequencies of apoB C7673T polymorphism in cerebral hemorrhage patients with family history, first-degree relatives, second-degree relatives, third-degree relatives, the sporadic cerebral hemorrhage patients and the control group were 0.176, 0.136, 0.058, 0.048, 0.081 and 0.040, respectively. (2) The allele T frequencies of apoB C7673T polymorphism in CHFH patients and their first-degree relatives were significantly higher than that of the control group (P< 0.01, P< 0.01), while there was no significant difference among second-degree relatives, third-degree relatives and control group (P> 0.05). And the allele T frequency of apoB C7673T in CHFH patients was significantly higher than that of sporadic cerebral hemorrhage patients (P< 0.05). (3)In CHFH patients and sporadic cerebral hemorrhage group, the levels of TC and LDL-C of the TC genotype were significantly higher than those of the CC genotype, while the level of HDL-C in the TC genotype was significantly lower than that of the CC geneotype (P< 0.05).

CONCLUSION(1)The allele T of apoB C7673T polymorphism may be related to cerebral hemorrhage with family history. (2) The allele T of apoB C7673T polymorphism may increase the susceptibility of cerebral hemorrhage by changing blood lipid levels.

Adult ; Aged ; Apolipoproteins B ; genetics ; Cerebral Hemorrhage ; blood ; genetics ; Cholesterol ; blood ; Cholesterol, HDL ; blood ; Cholesterol, LDL ; blood ; Female ; Gene Frequency ; Genotype ; Humans ; Lipids ; genetics ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Triglycerides ; blood

OBJECTIVETo assess the association between I405V and D442G polymorphisms of the CETP gene with cerebral hemorrhage (CH) and a related lipid profile among ethnic Han Chinese from Changsha.

METHODSA case-control study was carried out, which enrolled 170 cerebral hemorrhage patients and 191 ethnicity-, age- and sex-matched health controls. Polymerase chain reaction-restricted fragments length polymorphism (PCR-RFLP) was used to determine the polymorphisms. Lipid profile was determined by means of oxidase method. Statistic analyses were performed with SPSS 16.0.

RESULTSNo significant difference was found in the CETP gene I405V and D442G genotypes and allelic distribution between the CH patients and controls (P>0.05). There was no association between CETP gene I405V polymorphism and lipid profile in both groups (P>0.05). CH patients with DG genotype of the D442G polymorphism had higher TC and low density lipoprotein-cholesterol (LDL-C) levels than those with a DD genotype(P<0.05).

CONCLUSIONCETP gene I405V polymorphism may not be associated with CH among ethnic Han Chinese from Changsha, while the D442G polymorphism of the CETP gene may be associated with TC and LDL levels in the same population.

Adult ; Aged ; Asian Continental Ancestry Group ; ethnology ; genetics ; Base Sequence ; Case-Control Studies ; Cerebral Hemorrhage ; blood ; ethnology ; genetics ; China ; ethnology ; Cholesterol Ester Transfer Proteins ; genetics ; metabolism ; Cholesterol, HDL ; blood ; Female ; Humans ; Lipids ; blood ; chemistry ; Male ; Middle Aged ; Molecular Sequence Data ; Mutation, Missense ; Polymorphism, Single Nucleotide

OBJECTIVETo establish a multiplex RT-PCR-based reverse dot blot hybridization technique to detect influenza viruses.

METHODSObtain the HA nucleotide sequences of seasonal influenza H1N1, seasonal influenza H3N2, influenza H1N1 and human avian influenza H5N1 from GenBank. Design primers in conservative district and probes t in high variable region respectively, after analyzing the HA nucleotide sequences of influenza virus through the Vector NTI 9.0. Establish and optimize multiple RT-PCR system by comparing amplification efficiency and specificity at different primer concentrations. Establish the reverse dot hybridization system after optimizing the concentration of probes. To compare the sensitivity and specificity of this technique and the general RT-PCR Method through extracting the viral RNA of the mentioned influenza virus which are to be the reference substance.

RESULTSSuccessfully establish a multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses. This technique is 100-1000 times more sensitive than gel electrophoresis method, and it has a good specificity.

CONCLUSIONSuccessfully established multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses.

Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; Influenza A Virus, H3N2 Subtype ; genetics ; isolation & purification ; Influenza A Virus, H5N1 Subtype ; genetics ; isolation & purification ; Influenza, Human ; diagnosis ; virology ; Nucleic Acid Hybridization ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Objective To investigate the relationship between C1773T polymorphism of LDL receptor gene (LDLR) and cerebral hemorrhage and the impact of C1773T polymorphism of LDLR on the levels of serum lipids in Chinese Han in Changsha, Hunan province. Methods Two hundred seventy-three cerebral hemorrhage patients and 140 normal controls were recruited in the present study. The C1773T polymorphism of LDLR was analyzed by SNaPshot and direct DNA sequencing. The triglyceride (TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) levels were examined using oxidase method. Results The CC, CT and TT genotype frequencies of LDLR polymorphism were 0.703/0.278/0.019、0.707/0.250/0.043 and the allele C and T frequencies of LDLR polymorphism in the cerebral hemorrhage group and the control group were 0.842/0.158,0.832/0.168 respectively. The differences in genotype and allele frequencies of LDLR polymorphism were no significant between cerebral hemorrhage group and the control group (P>0.05). There were no significant differences in the levels of lipids among the CC, CT and TT genotype in either cerebral hemorrhage group or the control group (P＞0.05). Conclusions The LDLR-C1773T polymorphism may not be associated with cerebral hemorrhage nor be related to hyperlipemia in Chinese Han in Changsha.

To study the diagnostic method of slight viral myocarditis in the field of forensic pathology, slight viral myocarditis model was induced in Balb/c murine by coxsackie virus B3. Organs of hearts, livers, spleens, lungs and kidneys were examined through routine pathological methods. Pathological changes at different levels of these organs were observed. The results indicated that viral myocarditis was a kind of disease with multiple organ alterations and that the pathological observation and comprehensive analysis of multiple organs was one of the useful methods for diagnosing slight viral myocarditis.
Animals ; Coxsackievirus Infections/pathology* ; Female ; Forensic Medicine ; Male ; Mice ; Mice, Inbred BALB C ; Myocarditis/virology*

OBJECTIVE: To explore the association between apolipoprotein AI (ApoAI) gene rs12721026 polymorphism and cerebral hemorrhage (CH) in Changsha Han population, and to evaluate the effect of rs12721026 polymorphism on plasma lipid levels. METHODS: A total of 273 patients with CH and 140 healthy controls were collected. The rs12721026 polymorphism of ApoAI was analyzed by SNaPshot genotyping analysis and DNA sequencing. The total cholesterol (TG), triglyceride (TC), HDL-C and LDL-C were examined by oxidase method. RESULTS: There was no significant difference in the genotype and allele frequencies of rs12721026 polymorphism between the CH group and the control group (P>0.05). Both in the CH group and in the control group, the level of HDL-C of the TT gene type of rs12721026 was significantly higher than that of the GT/GG gene type (P<0.05). There was no significant difference in the levels of TG, TC and LDL-C among different subgroups of gene types. CONCLUSION There may be no association between apoAI gene rs12721026 polymorphism with CH in Changsha Han population, which may still influence the HDL-C levels.
Apolipoprotein A-I ; genetics ; Asian Continental Ancestry Group ; Case-Control Studies ; Cerebral Hemorrhage ; blood ; genetics ; Cholesterol ; blood ; Gene Frequency ; Genotype ; Humans ; Lipids ; Polymorphism, Single Nucleotide ; Sequence Analysis, DNA ; Triglycerides ; blood

OBJECTIVETo explore the value of the POSSUM scoring system in predicting postoperative morbidity and mortality of pancreatoduodenectomy (PD).

METHODSTwo hundreds and sixty-five consecutive PDs were performed between January 2005 and December 2007. POSSUM scores which relied on 12 physiologic and 6 operative variables were prospectively calculated for each case. Expected morbidity and mortality were estimated based on POSSUM scores and were compared with observed morbidity, which were diagnosed according to the Clavien complication scheme and domestic reference criteria respectively, and mortality.

RESULTSPhysiologic scores of 265 cases ranged from 12 to 24,the mean was 15. Operative scores ranged from 14 to 24, the mean was 17. The overall POSSUM scores ranged from 0.24 to 0.88. Average expected morbidity was 43.8%, expected cases were 116. Observed morbidity rate was 39.6% (105/265). The expected and observed morbidities and cases had no significantly differences. All patients were classified to 1 of 4 strata based on their individual POSSUM scores and subsequent risk of morbidity. Predictive value was the highest when scores ranged from 0.4 to 0.8. POSSUM exhibited less predictive value for mortality, but if POSSUM was more than 0.5, it was useful for mortality predicting.

CONCLUSIONSPOSSUM scoring system has high value for predicting the risk of morbidity in PD and can be helpful in guiding surgery and postoperative management decisions.

Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Pancreaticoduodenectomy ; mortality ; Postoperative Complications ; Prospective Studies ; Risk Assessment

Objective To explore the efficacy and safety of autologous peripheral blood stem cell transplantation (APBSCT) in the treatment of systemic lupus erythematosus.Methods Nine patients with systemic lupus erythematosus were enrolled in this study.Patients were given cyclophosphamide and granu- locyte colony-stimulating factor(G-CSF)as the mobilization regimen.Urine was alkalinized and hydrolyzed to protect the function of the heart,liver and kidney of the patients.A CS3000 Plus blood cell separator was used to collect peripheral blood stem cells,which were preserved in liquid nitrogen.Two to five days before the administration of the stem cells,the patients were pretreated with intravenous injection of cyclophos- phamide (50 mg?kg~(-1)?day~1) for 4 consecutive days and antithymocyte globulin (ATG,2.5 mg?kg~(-1)?day~1) for 3 consecutive days.Granulocytes were recoverd by G-CSF stimulation.Then,the peripheral blood stem cells were reinfused.Therapeutic effect was evaluated by assessment of alteration of clinical manifestation (skin erythema),levels of proteinuria and antoantibodies,hematopoietic reconstitution and occurrence of transplantation related complications.Results After transplantation,all patients had been successfully en- grafted.The time for peripheral leucocyte count to reach 1.0?10~9/L was 7～15d;the time for platelets to reach 20?10~9/L was 0～21 d.The skin erythema resolved in all patients;proteinuria decreased to normal level and the autoantibodies became negative in most of the patients.Serum sickness-like response occurred in all patients,renal and heart failure in 1 patient,hemorrhagic cystitis in 3 patients,psychiatric disorders in 1 patient,candidal infection in 1 patient.Conclusion One-year follow up suggests that autologous stem cell transplantation is markedly effective and relatively safe for systemic lupus erythematosus.However,the duration of remission remains to be investigated in a long-term follow up study.