This paper recommends the early diagnosis of epidemic hemorrhagicfever with immunoenzymatic direct staining (IDS) to study the patient'speripheral white blood cells. The samples of blood smears from 63 patientswere examined under the microscope with this metbod. In 54 patients, the results were positive (85. 71%). We systemically observed 10 blood sam-ples from the patients in different courses of the disease, and found thatduring the period of recovery the results were negative. This method wasalso used to observe the blood smears from both 22 patients infected withother viruses and 20 healthy persons, and a negative result was obtained.It was demonstrated that IDS had stronger specificity. The advantages ofthis method are that blood samples are easily taken, carried and preserved,and that the experimental method is simple. Thus, IDS is of practical signifi-cance in preventing and treating the disease.

Objective To investigate the changes in skin microcirculatory perfusion during induction of general anesthesia and the effects oftwo fluid therapy regimens in patients undergoing abdominal surgery.Methods Thirty-six ASA Ⅰ or Ⅱ patients aged 18-64 yr scheduled for elective major abdominal surgery were randomized to receive either 6% hydroxyetlayl starch(130/0.4)7 ml/kg(HES group,n=18)or lactated Ringer's solution 7 ml/kg(RL group,n=18)for compensatory intravascular volultne expansion(CVE)before tracheal intubation.Meanwhile both groups received continuous intravenous infusion of RL at a of 8 ml·kg~(-1)·h~(-1).Tracheal intubation was performed at 40 min after the onset of infusion.Anesthesia was maintained with with sevoflurane,remifentanil and rocuronium.Operation was started at 20 min after tracheal intubation.The microcirculatory perfusion was measured on forehead skin by using Doppler perfusion imaging system(LDPI)PI)at the onset of fluid infusion(T_0,baseline),the end of endotracheal intubation(T_1)and the onset of skin incision(T_2).Rwsults The MAP,HR,blood gases and body temperature were within the normal during the experiment and there was no significant difference between the 2 groups.The skin microcirculatory perfusion and CVP at T_1 were significantly higher in group HES than in group RL(P<0.05 or 0.01).Compared with the baseline value at T_0,the skin microcirculatory perfusion at T_1 was significantly increased in group HES(P<0.01),but there was no significant change in the skin microcirculatory perfusion at T_1 in group RL(P>0.05),the skin microcirculatory perfusion at T_2 was singificantly decreased in both groups(P<0.01),and CVP and PaO_2/FiO_2 at T_(1.2) were significantly increased,while Hb at T_(1.2) was significantly decreased in both groups(P<0.05).The skin microcirculatory perfusion in both groups was significantly lower at T_2 than at T_1(P<0.01).Conclusion The infusion of 6% HES 130/0.4 can improve the skin microcirculatory perfusion and the effect is better than that of RL during induction of general anesthesia in patients seheduled for abdominal surgery.

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Efficient gene delivery and sustained gene expression are required for successful human gene therapy.Although viral vectors are considered the most efficient vehicles for gene transfer,currently available viral vectors have not fully achieved these two requirements.Lentiviral vectors(LVs)can integrate into host chromosomes,allowing long-term gene expression,in addition,these vectors are non-toxic and minimally immunogenic since no viral genes are encoded in the vector genome,but are still limited to in vitro or ex vivo gene delivery because of their relatively low titers using transient transfection experiments.In order to develop an efficient transient transfection method for large-scale production of high titer lentiviral vector stocks,a minimal lentiviral vector producing system based on vaccinia virus that synthesizes T7 RNA polymerase was developed.BHK21 was co-transfected by three main plasmids containing the transducing plasmid pVECRNA,the packaging plasmid pGAGPOL and the envelope plasmid pVSVG,and thereafter infected with the vaccinia vTF-3 containing bacteriophage T7 RNA polymerase gene using Lipofectamin2000TM.After 4 days,the culture supernatant of lentiviral vectors was collected,the RNA from the supernatant was examined by the RT-PCR,the protein from the supernatant was examined by Western blot,and the supernatant was used to transfect normal 293T,HepG2 and Vero,which were observed by the immunofluorescence microscopy.The type of cell lines,plasmids dosage and the MOI(the proportion between cell numbers and virus copies)were considered so critical to the output of this system that 3?3?3 factorial design was used to explore the yield optimization of this system.As judged by the results of RT-PCR and the Western blot,lentiviral vectors were found in the culture supernatant;as judged by immunofluorescence with microscopy,293T,HepG2 and Vero which were transfected by the supnantant expressed the report protein-green fluorescent protein(GFP),the results confirmed the valid infectivity of the lentiviral vector produced by the system.Eventually,the best titers of lentiviral vector stocks was up to 1.3?108 tu/ml,which is one order of magnitude higher than the output of classical manufacture system.The new poxviral/lentiviral hybrid system for efficient lentiviral vector production was initially established.It provides the basis for the future development of industrial application.

Chinese materia medica (CMM) as well as its interrelated industries has huge exploitable potential and developing prospect At present, it has become one of the directions with the worldwide exploitation of new medicine and development in medicinal industry The establishment of quality standard system of CMM in China is one of the key problems that the modernization of them should be first of all resolved In order to ensure the accuracy, credibility, comparability and mutual recognition of data examined by each laboratory, our research should be based on the international theory and method of the traceability Our government and CMM administration department should strengthen the esablishment and promulgation of criteria, laws, regulations of CMM and its interrelated industries, so as to make the quality assurance system approach to the international standard, and to protect our economic benefits, people's health and most consumers' interests from being affected

Objective To express lactate dehydrogenase (LDH) gene of Plasmodium falciparum FCC1/HN in the E. coli TG1 and analyse its immunocompetence. Methods The LDH gene of the P. falciparum was specifically amplified by polymerase chain reaction, and the recovered gene fragment was cloned into pGEX 4T 1 vector for expression of fusion protein with glutathione S transferase(GST). The recombinant plasmid was transformed into the E. coli TG1. Four mice (Kunming strain) were immunized with purified expressed protein(antigen) and the polyclonal antibodies were collected. The immunocompetence of recombinant protein was analysed by ELISA and Western blot. Results The LDH gene of P. falciparum was successfully expressed in the E. coli TG1. The expressed protein exhibited a specific reaction with immune sera obtained from rabbits immunized with P. falciparum . The specific humoral responses were induced in mice and the titer of the specific antibody was 1∶16 by two dimensional diffusion assay. Conclusion The LDH gene of P. falciparum has been successfully expressed in the E. coli TG1 and the expressed protein has high antigencity.

Objective Measure the variation trend and the problems of bed allocation and its equity at primary medical institutions and provide theoretical basis for optimizing the policy of health resources allocation for the government.Methods Analysis of the distribution and equity of the inpatient beds at primary medical institutions in terms of geographical area by means of the survey of regular overall statistics forms and the methods of Gini coefficient and Theil index.Results From 2009 to 2012,the number increase of beds at such institutions geographically was 7.72%,while that in eastern,central and western regions of China were respectively 5.35%, 3.97% and 27.88%.The number increase at community healthcare centers was 46.58%,while that in eastern,central and western regions of the country were respectively 18.09%,29.57%和 81.53%.The number increase at township hospitals was 2.33%,while that in eastern,central and western regions were respectively - 3.83%,- 0.43% and 22%.The Gini coefficient of bed allocation at such institutions was beyond 0.6,while that in eastern, central and western regions were respectively under 0.3,0.3-0.4 and 0.6 above.The Gini coefficient of bed allocation at community healthcare centers fell from above 0.4 to under 0.4.The Gini coefficient in the western regions increased from 0.3-0.4 to over 0.4.The variation trend of township hospitals was similar with the overall conditions at such institutions.The intra-regional Theil indexes and their contribution rates of bed allocation at such institutions were higher than those of inter-regional. Conclusions The overall variation trend of bed allocation at such institutions was on the rise,yet its variation trend of equity remains unchanged.The bed allocation and its equity at such institutions were higher than those at township hospitals.The growth rate in western regions was the highest,but the absolute number was the lowest.Moreover,the equity of bed allocation in such regions was not only the lowest,but also was found rising.The influence of bed allocation in intra-region terms was large,while its equity was not only lower but falling as well.

Methylene blue injection has a stronger direct inactivation on VZV in vitro. When the injection was diluted from 1:16 to 1:128, which was obvious (P＜0.01 and P＜0.05). The MIC was 1:222, the IC50 was 1:135 and IC90 was 1:77. The results of microcellculture method showed when the injection was diluted from 1:16 to 1:64, it also effectively inhibited the proliferation of VZV in WISH continuous cell-lines (P＜0.01 and P＜0.05). The MIC was 1:95, the IC50 was 1:45 and the IC90 was 1:21.

Purpose To ana1yze the expression features of 5-F1uorouraci1(5-FU)metabo1ic enzyme thymidy1ate synthetase(TS),thy-midine phosphory1ase( TP),dihydropyrimidine dehydrogenase( DPD)and its re1ationship with c1inicopatho1ogica1 factors and progno-sis in co1orecta1 cancer,in order to further exp1ore its potentia1 significance in guiding co1orecta1 cancer chemotherapy. Methods Es-tab1ishment of a tissue microarray containing 72 patients with co1orecta1 cancer,and 56 norma1 tissue( dista1 cut edge tissue near carci-noma)was used to detect TS,TP,and DPD by immunohistochemistry,and to ana1yze its re1ationship with c1inicopatho1ogica1 factors and prognosis of co1orecta1 cancer through statistica1 method. Results The expression of TS in co1orecta1 cancer was 1ower than that in norma1 tissue(P=0. 876),which was associated with TNM(P=0. 043)and positive1y corre1ated with patients’overa11 surviva1(P=0. 027),the expression of TP in co1orecta1 cancer was higher than that in norma1 tissue(P=0. 315)that was associated with 1ymph node metastasis(P=0. 009)and negative1y corre1ated with the prognosis of patients(P=0. 040),DPD expression in co1orecta1 canc-er was higher than that in norma1 tissue(P=0. 071),which was re1ated to the histo1ogic type(P=0. 029). Overa11 surviva1 was sig-nificant1y shortened in co1orecta1 cancer with DPD high expression( P=0. 011). Conclusions TS,TP and DPD might be app1ied as important index of prognosis in co1orecta1 cancer patients using 5-FU adjuvant chemotherapy. The expression of TS re1ated c1ose1y with the c1inica1 stage is a bio1ogica1 marker of tumor progression. TP expression is c1ose1y re1ated to 1ymph nodes metastasis and recur-rence,which is an important factor of postoperative recurrence and metastasis.

Objective:To study the effects of tamoxifen on proliferation and ER expression of human hepatocellular carcinoma cells.Methods:HepG2 cells were treated with tamoxifen at different concentration and different action time.MTT was used to determine the suppression rate of human hepatocellular carcinoma cell.The effects of tamoxifen on human hepatocellular carcinoma cell ER performance were observed by immunohistochemistry.Results:Tamoxifen inhibited the proliferation of human hepatocellular carcinoma cells and suppressed human hepatocellular carcinoma cell ER performence.Conclusions:Tamoxifen may suppress human hepatocellular carcinoma cell proliferation ER performance.