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.

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.

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.

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.

Objective To observe the effects and study the underlying mechanism of siRNA targeting PARP1 on the proliferation of androgen independent prostate cancer PC3 cell line.Methods Three specific siRNA sequences targeting PARP1 were designed and synthesized.And two sequences which had better interfering effect on the expression of PARP1 were evaluated and selected through lipofectamine transfection,RT-PCR and Western Blot.The effect of PARP1 silencing on the proliferation of PC3 cells was observed with MTS assay and the levels of the phosphorylation of Akt and GSK3β were detected by Western Blot.Results Compared to the blank control group,the transfected group with the negative control sequence had no significant impact on the expression of PARP1,however the transfected group with siRNA-1706,-2003 or-2907 could significantly suppress the mRNA and protein expression of PARP1.The mRNA inhibition rate reached to(52.07 ± 4.65)％,(44.38 ± 9.15)％ and(22.05 ± 6.65)％,respectively;and the protein inhibition rate reached to(86.86 ± 4.94)％,(83.30 ± 7.18)％ and(63.05 ± 10.19)％,respectively.The siRNA-1706 and-2003 could significantly inhibit the proliferation of PC3 cells;the inhibition rate was(38.93 ± 3.87)％ and(34.93 ± 1.21)％.And they also could down-regulate the intracellular levels of phosphorylated Akt and GSK3β in PC3 cells.Conclusion PARP1-targeted siRNA can significantly suppress the expression of endogenous PARP1 and inhibit the proliferation of PC3 cells,which is related to the inhibition of Akt activity and the activation of GSK3 β.

Objective To evaluate the feasibility and efficiency of laparoscopic-guided radiofrequency ablation on advanced prostate cancer.Methods From March 2003 to December 2008,a total of 6 previous prostate cancer patients who had been diagnosed with pathological results were treated by laparoscopic-guided radiofrequency ablation.All patients underwent pre-and post-operative IPSS,serum PSA,MRI and normal blood biochemistry examination.The treatment outcome,surgery-related complications were also recorded.Results All operations were successfully completed,no serious intra-and post-operative complications happened.Although there was no significant difference of IPSS between pre-operative [ ( 19.05 + 4.28 ) scores ] and 1 month after operation [ ( 19.87 + 5.72) scores ],but there were significantly decreased in 3 months [ (9.45 ± 2.03 ) scores ] and 6 months [ (6.18 + 1.79) scores ] after operation (P ＜0.05).Also being followed up to 6 months after operation,the serum PSA was significantly decreased compared with the pre-operative value [from(24.80 ± 14.56) μ g/L reduced to( 13.79 ± 7.76) μ g/L](P＜0.05).Conclusion Laparoscopic-guided radiofrequency ablation on advanced prostate cancer is safe and feasible,and can be used as an effective treatment in selective cases.

Objective To compare the efficiency between mini-percutaneous nephrolithotomy (MPCNL) and extracorporeal shock wave lithotripsy (ESWL) for monotherapy of renal calculi in infants less than 3 years.Methods Forty-six infants were treated with either SWL (22 infants) or MPCNL monotherapy (24 infants).The mean age was (22.6 ± 8.7) months vs (23.5 ± 6.6) months and the stone size was (21.4 ± 3.5) mm vs (21.7 ± 1.7) mm,and there were no significant difference.Results For MPCNL,mean operating time was (76.2 ± 23.4) min and mean hospital stay was (14.13 ± 5.8) d.The stone-free rates were 84.0％ (21/25) after first session and 96.0％ (24/25) after second-look procedure.Postoperative fever happened in 4 (16.0％) cases.Hemoglobin drop was (8.5 ± 4.4) g/L and no one needed blood transfusion.For ESWL,the stone-free rate were 31.8％ (7/22) after first session and 86.3％ (19/22) after second session for 11 infants (50.0％).Mean hospital stay was (6.6 ± 2.3) d and 10 cases (45.5％)had complications.Hemoglobin drop was (10.6 ± 12.7) g/L.MPCNL was lower than ESWL in complications rate and re-treatment rate,and the stone-free rate was higher,but the hospital stay and operation time was longer (P ＜ 0.05).GFR revealed improve or stable after operation in both groups.Conclusions For a higher success rate,lower complication rate and re-treatment rate,MPCNL was an effective option for the management of relatively larger stones in children (even in infants).

Objective To determine the median effective doae (ED50) for motor block after intrathecal ropivacaine and bupivacaine. Methods Sixty ASA Ⅰ or Ⅱ patients, aged 18-64, weighing 46-75 kg, undergoing elective urological surgery under combined spinal-epidural anesthesia, were randomized into 2 groups ( n = 30each) receiving intrathecal 0.5% ropivacaine and 0.5% bupivacaine respectively. The ED50 was determined by up-down sequential allocation. The initial dose was 4 mg. Each time the dose increased/decreased by 1 mg. Efficacy was determined by the occurrence of any motor block in either lower extremity (modified Bromage scale > 0)within 5 or 10 min after the spinal injection. Results The intrathecal ED50 for motor block was 6.68 mg for ropivacaine (95% confidence interval 6.27-7.13 mg) and 4.07 mg for bupivacaine (95% confidence interval 3.56-4.47mg) . The relative motor blocking potency ratio was ropivacaine/bupivacaine 0.61. Conclusion The potency of intrathecal ropivacaine is lower than that of bupivacaine for motor block.

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.

Objective To evaluate the efficacy and advantages of the technique by combined PCNL and retrograde intrarenal surgery (RIRS) in a second stage to treat the complex renal stones in solitary kidney cases.Methods PCNL most with a single 18-24 F tract was performed first and RIRS was carried out at a second stage in solitary kidneys of 21 patients,of which congenital in 14.3％ (3 cases),contralateral nephrectomy in 42.8％ (9 cases),and functional solitry kidneys in 42.8％ (9 cases).Of the 21 patients,the average age was 45 years with 15 males and 6 females.The stones were 8 multiple,6 partial staghorn,and 7 complete staghorn with a mean size of 4.6 (3.8-6.8) cm.Results Of the 21 PCNL cases,a single tract,double,triple tracts were established in 18 (85.7％),2 (9.5％),1 (4.8％) cases,respectively.Mean operation time of PCNL was 95 (45-175) min.After 1 day of PCNL,all case had residual stones with a mean size of 1.9 (1.0-3.5) cm.Two case occurred fever after PCNL and one case presented bleeding resolved by selective renal artery embolization.The mean operation time of RIRS was 72 (35-95) min.Stone-free rate after RIRS was 85.7％ (18/21).The final stone free rate increased to 95.2％ (20/21) after one case received a second-look PCNL and two cases accepted ESWL.Two cases occurred fever and steinstrasses after RIRS resolved by rigid ureteroscopy.At the 3 months follow-up,renal function became stable,improved and worse in 71.4％ (n=15),23.8％ (n=5),and 4.8％ (n=l) of patients.Conclusions PCNL combined with RIRS could be an effective and safe option for complex stones in solitary kidneys with less bleeding,reduced tracts,minor complications and good renal function preservation.