The pandemic outbreak of influenza has been started from Mexico in 2009 to 70 countries during 2 months. On 11th of June , WHO announced influenza pandemic alert level rose to the highest level 6, which means the first influenza pandemic in 21st century is coming. Till 6th of July, 94 512 confirmed cases from more than 120 countries and areas were reported, including 429 cases were died. The genetic fragment of swine, poultry sources and human influenza viruses are contained in this strain, A/H1N1 influenza virus, of the pandemic. It is of great significance of studying the genetic reassortment, evolution and its biological characteristics of this virus strain to prevent and control the pandemic. At present, the genetic evolution of strain has been identified, and the potential biological characteristics have been analyzed by genetic traits, however, clinical manifestation should be further concerned, and the tendency of influenza pandemic and genetic changes need to be monitored closely. The complexity of influenza virus ecosystems, mutation of genome, and easy to preserve in "Nature Gene Pool" and reassortment, make the influenza pandemic inevitable. We should face the threat of influenza pandemic, enhance the surveillance of influenza virus in ecosystems, strengthen the epidemiological investigation, develop the vaccines and drugs, and establish an effective public health security system, in order to reduce the destruction of the influenza pandemic.

Objective To study a case of severe hemolytic disease with the newborn induced by DⅣb type of RhD variant and to investigate its molecular mechanism .Methods Indirect Coombs test was performed to identify RhD blood type and detect antibodies against red blood cells (RBCs).RHD genes were analyzed by polymerase chain reaction-sequence specific primers (PCR-SSP) analysis.All of the 10 exons of RHD gene were sequenced .The Rhesus boxes were further analyzed to identify the homozygosis of RHD genes.Results The mother of the newborn was RhD positive carrying anti-D antibody.PCR-SSP analysis indicated that the RHD exons 7-9 were missing, although the sequences of other RHD exons were consistent with standard sequences .RHD zygosity test showed that the mother was RHD+/RHD-.The newborn was RhD positive with anti-D antibody in serum .The result of the direct antiglobulin test was also positive .The sequence of the RHD exons 1-10 of the newborn were identical with standard sequences .The genotype of the newborn was identified as RHD+/RHD+homozygote .Conclusion The mother bears a DⅣb genotype lac-king RHD exons 7-9 which is significantly different from the newborn .The anti-D antibodies in the mother might induce the severe hemolytic disease in the newborn .

Objective To explore the possibility of early definitive operation for enteric fistulas. Methods In 37 selected enterocutaneous fistula patients, early definitive operation was performed during laparotomy for treating peritonitis within 10 days of fistula formation. Thorough irrigation of peritoneal cavity, effective sump drainage, fibrin glue sealant to reinforce anastomosis, appropriate nutritional support, and administration of growth hormone were key elements of perioperative management. Results Among 37 patients, 35 of them recovered uneventfully. Fistula recurred in 2 cases postoperatively; one fistula closed after conservative management, and another patient died of advanced gastric cancer 3 months postoperatively. The fistula operative closure rate was 94.5%. No operative death. Conclusion Advances in perioperative management can promote the success rate of early definitive operation for enteric fistulas. It may become a challenge to the present strategy of enteric fistula management.

OBJECTIVE To determine the prevalence of post-operative infections in patients who underwent damage control laparotomy(DCL) with abdominal packing and to identify the risk factors,mortality and predominant pathogens.METHODS A retrospective study of postoperative infections and microbiology in patients who underwent abdominal packing as an adjunct of DCL to control coagulopathic hemorrhage over a 5 year period(Feb 2002-Feb 2007) were performed.RESULTS A total of 26 patients were studied.Pneumonia/lower respivatory tract infection was the prominant type of infection(57.7%),followed by bacteremia(50.0%),urinary tract infection(15.4%) and wound infection(15.4%).Of the 244 organisms isolated from various sites,the most frequently isolated bacteria were Pseudomonas aeruginosa(27.0%),Staphylococcus species(15.6%),Acinetobacter baumannii(13.9%),and Klebsiella species(11.1%).No statistical correlation was found between positive packs and postoperative infection(P=0.10) or death(P=1.00).Multivariate regression analysis revealed that pre-existing abdominal infection(OR=22.4,P=0.02) and increased number of surgical procedures(OR=3.69,P=0.05) were the independent risk factors for post-operative infections.CONCLUSIONS Patients who undergo DCL with packing have a high incidence of postoperative infections.Pathogens and distribution are same as acquired infections.Pre-existing abdominal infection and increased number of surgical procedures are the independent risk factors for postoperative infections in these patients.

Objective To evaluate the application value of PCR-reverse dot blot hybridization in the identification of Candida and the detection of Candida albicans drug-resistant genes.Methods The vaginal secretion samples from 285 patients with candidal vaginitis and 50 healthy women were collected.The identification of Candida species and their drug susceptibility were detected by the bioMérieux Yeast identification cards and MIC method(Zhengzhou Antu kit),respectively.The identification of Candida species and the mutation of Candida albicans,drug-resistant genes were also detected by the Shenzheng Yaneng test kit(PCR-reverse dot blot hybridization).The drug-resistant genes were also identified by PCR and nucleic acid sequencing.Based on the culture identification,MIC method and nucleic acid sequencing as the contrast methods,the sensitivity,specificity and accuracy of PCR-reverse dot blot hybridization in the identification of Candida species and the mutation detection of Candida albicans drug-resistant genes were evaluated.Results Compared with the bioMérieux Yeast identification method,the sensitivity,specificity,positive predictive value,negative predictive value and total coincidence rate of PCR-reverse dot blot hybridization for detecting six kinds of Candida species,including Candida albicans,Candida glabrata,Candida tropicalis,Candida parapsilosis,Candida krusei and Candida guilliermondii,were above 95％,96％,96％,98％ and 97％,respectively.There was no significant difference in detecting six kinds of Candida species between the two methods (x2 =0.44,0,0,0,0 and 0,respectively,P ＞ 0.05),and there was good consistency between them (Kappa ＞ 0.9).Compared with the MIC method,the sensitivity,specificity,positive predictive value,negative predictive value and total coincidence rate of PCR-reverse dot blot hybridization for detecting the drug resistance of Candida albicans were 98％,88％,98％,88％ and 96％,respectively.There was no significant difference in detecting the drug resistance of Candida albicans between the two methods (x2 =0.17,P ＞ 0.05),and there was good consistency between them (Kappa ＞ 0.8).The results of PCR-reverse dot blot hybridization in detecting the mutation sites of six kinds of Candida albicans drug-resistant genes were 100％ of coincidence with that of the nucleic acid sequencing method.Conclusion The PCR-reverse dot blot hybridization has high consistency with the culture method and the nucleic acid sequencing method in the identification of Candida species and the mutation detection of Candida albicans drug-resistant genes,which is more early and rapid than the traditional detection methods,and may be applied to the auxiliary diagnosis of vulvovaginal candidiasis (VVC).

Objective To explore the clinical effect of neoadjuvant chemo-radiation combined with thoracoscopic and laparoscopic three-incision esophagectomy for esophageal cancer.Methods The retrospective cohort study was conducted.The clinicopathological data of 111 esophageal cancer patients who underwent neoadjuvant chemo-radiation combined with thoracoscopic and laparoscopic three-incision esophagectomy in the Cancer Hospital of Chinese Academy of Medical Sciences between January 2010 and December 2016 were collected.Among 111 patients,68 with interval time between neoadjuvant chemo-radiation and thoracoscopic and laparoscopic three-incision esophagectomy ＜ 8 weeks were allocated into the ＜ 8 weeks group and 43 with interval time between neoadjuvant chemo-radiation and thoracoscopic and laparoscopic three-incision esophagectomy ≥8 weeks were allocated into the ≥8 weeks group.Patients underwent preoperative radiotherapy and chemotherapy with TP regimen,and then underwent selective thoracoscopic and laparoscopic three-incision esophagectomy.Observation indicators:(1) neoadjuvant chemo-radiation situations;(2) surgical and postoperative situations;(3) follow-up.Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival of patients and tumor recurrence or metastasis up to March 2017.Measurement data with normal distribution were represented as (x)±s and comparison between groups was analyzed using the t test.Count data were analyzed using the chi-square test or Fisher exact probability.Comparison of ranked data was done by the nonparametric test.Results (1) Neoadjuvant chemo-radiation situations:all the patients underwent neoadjuvant chemo-radiation,without severe adverse reaction.Number of patients with complete remission based on oncopathology were 34 in the ＜8 weeks group and 15 in the ≥ 8 weeks group,with no statistically significant difference between the 2 groups (x2=2.441,P＞0.05).(2) Surgical and postoperative situations:all the patients underwent successful thoracoscopic and laparoscopic three-incision esophagectomy,with negative surgical margins.Operation time,volume of intraoperative blood loss,number of lymph node dissected,time of postoperative intrathoracic drainagetube removal,time of postoperative neck drainage-tube removal,hoarseness,upper gastrointestinal hemorrhage,anastomotic fistula,respiratory complication,pleural effusion and empyema,cardiovascular complication,incision abnormal healing,death within postoperative 30 days and duration of hospital stay were (354±103)minutes,(400± 76)mL,19±4,(11±4)days,(4.9±1.6)days,5,1,12,3,6,5,8,0,(19± 17) days in the ＜ 8 weeks group and (343±92) minutes,(392±51)mL,19±3,(12±6)days,(4.5±1.0)days,2,0,7,5,3,4,3,2,(18± 11) days in the ≥ 8 weeks group,respectively,with no statistically significant difference between the 2 groups (t =1.080,0.569,0.326,1.223,1.286,x2=0.029,0.035,1.114,0.000,0.000,0.246,t=0.315,P＞ 0.05).(3) Follow-up:90 of 111 patients were followed up for 3-82 months,with a median time of 25 months,including 55 in the ＜8 weeks group and 35 in the ≥8 weeks group.During follow-up,death and tumor recurrence were detected in 9,11 patients in the ＜8 weeks group and 6,11 patients in the ≥ 8 weeks group,respectively.Conclusion Neoadjuvant chemo-radiation combined with thoracoscopic and laparoscopic three-incision esophagectomy is safe and effective,and it doesn't increase the perioperative risks based on preoperative 8-week interval time.

Objective To explore the effects of protein kinase C alpha(PKC?) cDNA on the expression of genes of multidrug resistance (MDR) in renal cell carcinoma(RCC) 786-0 cell line.Methods LR/BR recombination reactions were used to generate mammalian expression vectors of PKC? cDNA with C-terminal fused green fluorescent protein(GFP),and vectors were transfected into human RCC cell with Lipofectimine 2000.Western blot method and inverted fluorescent microscope were used to determine the expression of PKC? in RCC cells transfected by PKC? cDNA.RT-PCR was used to determine the expression of MDR-related genes MDR1,MRP1 and LRP in RCC cells transfected by PKC? cDNA.Results After transfection of 786-0 cells with pcDNA-DEST47-PKC?-GFP vectors,the results of Western blot showed that PKC? was highly expressed in human RCC 786-0 cells;and inverted fluorescent microscopy showed that GFP was highly expressed in RCC 786-0 cells.The results of semi-quantitative RT-PCR analysis showed that the expression level of MDR1 was higher in RCC cells transfected by PKC? cDNA than in RCC cells.Conclusions The expression of MDR1 mRNA in 786-0 cell line can be up-regulated by PKC? cDNA,which suggests PKC? cDNA can induce the increase of MDR in renal cell carcinoma.

Classic problem-based learning ( PBL ) was applied in a medical microbiology course.The results of questionnaire survey showed most of the students were satisfied with the whole learning program.They thought the case was appropriate,and the discussing could increase their understanding of the knowledge.Based on the problems faced during the application and the students' feedback,the duty of the tutors,summary of the learning topic,arrangement of the course,application of different teaching methods,and selection of the tutors were discussed in detail.

Objective To summarize and investigate the lethal factor of acute obstructive suppurative cholangitis (AOSC).Methods The clinical data of 56 patients with AOSC were retrospectively analyzed.Results Six cases died,5 cases with acidosis,5 cases with thrombocytopenia and 5 cases with temperature change obviously,4 cases with heart,lung and kidney disease or diabetes,5 cases with operation and operation time ≥ 150 min,5 cases with from onset to treatment time ≥72 h.Eighteen cases of elderly patients ≥70 years old,4 cases died.The patients whose age≥70 years,temperature ≥39 ℃ or ＜ 36 ℃,combined with acidosis,platelet counts ≤6.0 × 1012/L,with heart,lung,kidney diease or diabetes,time of anesthesia and operation ≥ 150 min and from onset to treatment time ≥72 h had higher death rate (P ＜ 0.05).Conclusion Age,obvious temperature abnormalities,significantly platelet decrease,with heart,lung,kidney diease or diabetes,acidosis,long time of anesthesia and operation and from onset to treatment time ≥ 72 h are the lethal factor of AOSC.

Objective:To study the rule of spontaneous behavior and to explore the effect on emotion of mice peripherally infected with influenza A WSN33 virus( H1N1).Methods: Mice were intranasal inoculated with H1N1 WSN33 or saline.Then mice bodyweight change,and total distance movement,average movement speed distance in the central area and feces in the open field test in 5 minutes were recorded in two weeks.Results: Mice following WSN33 infection bodyweight declined sharply until day 7 post-inoculation,and mice bodyweight recovered from influenza infection at day 8 post-inoculation.Total distance movement of mice following H1N1 WSN33 infection decreased in the open field test,and difference of the reduction was significant from day 5 to day 10 post-inocu-lation.The average movement speed had no statistical difference.The range of numbers of fecal grains was large, and they were no significant difference.Conclusion:The total distance movement decreased,but average movement speed did not change following mice infected with H1N1 WSN33.They told us that mice infected with H1N1 WSN33 had anxiety,depressed and nervous emotion which is more evident in acute stage and early recovery stage,whereas the change of the nervous emotion was small and not obvious.