Objective: To investigate the evolutionary relationship of the hemagglutinin (HA) gene of novel influenza virus A/H1N1 in 2009 pandemic with the HA genes of A/H1N1 viruses isolated in different parts of the world previously. Methods: The sequences of the HA gene of the novel A/H1N1 strain and the reference sequences of human, swine, and avian influenza A viruses were retrieved from NCBI. MEGA 4.0 software was employed to align, blunt nucleotide sequences, and construct phylogenetic tree. The deduced amino acid sequences of the HA genes of novel influenza virus A/H1N1 were compared with those of the A/H1N1 isolates in North America, Europe, and Asia. Results: Phylogenetic tree of the HA genes of A/H1N1 strains worldwide showed that the HA genes of novel influenza virus A/H1N1 in 2009 shared a high homology with those of the 7 human A/H1N1 influenza viruses isolated in North America during 1976 to 2007, and shared a low homology with those of the human influenza viruses A/H1N1 isolated in Europe and Asia. Phylogenetic tree of the HA gene between different species showed that the HA genes of novel influenza virus A/H1N1 in 2009 had a close evolutionary relationship with those of the two swine A/H1N1 strains isolated in 1998 and 2007 in North America, but a distant evolutionary relationship with those of swine and avian A/H1N1 isolated in Europe and Asia. Alignment of amino acid at important antigenic sites showed that the HA gene of the novel A/H1N1 strains shared important antigen sites with the swine A/H1N1 influenza viruses isolated in North America, and did not share with the swine A/H1N1 influenza viruses isolated in Europe and Asia or the human A/H1N1 influenza vaccine strains. Conclusion: The HA genes of the novel influenza virus A/H1N1 might originate from swine A/H1N1 influenza viruses in North America after a long time evolution and the reassortment with fragments of human A/H1N1 in the area, and the current A/human/H1N1 influenza vaccine may not be effective for the novel A/H1N1 virus.

Objective To investigate the clinical outcome of post-traumatic tibial osteomyelitis treated by induced membrane technique combined with the antibiotic cement-coated locking plate.Methods A restrospective case series analysis was made on 71 cases of post-traumatic tibial osteomyelitis treated by induced membrane technique from September 2014 to September 2014.There were 55 males and 16 females, aged 18-60 years(mean, 37.4 years).Mean length of bone defect following debridement was 4.5 cm(range, 4-11 cm).Antibiotic cement-coated locking plates were used to provide the stability of the bone defects after one-stage debridement.Bone grafting and exchanging the plates with ntramedullary nails were done during the second-stage surgery.Parameters were monitored dynamically after operation to evaluate infection recurrence and bone healing, including C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), clinical features(sinus formation, redness and swelling, warmth and pain) and X-ray films of the involved limbs.Results All patients were followed up for 13-25 months (mean, 19.5 months).There was no sign of recurring infection and CRP and ESR were normal in all cases after the first stage surgery.However, four cases (6%) had recurrent infection with sinus exudates and their CRP and ESR were abnormal after the second stage surgery, and further treatments were carried out on these patients.Follow-up showed bony union in all patients within mean 5.5 months (range, 4-6 months).Conclusion Induced membrane technique with antibiotic cement-coated locking plates for treatment of post-traumatic tibial osteomyelitis can shorten bone healing time and decrease infection recurrent rate without obvious impact on osteogenic activity.

Objective To investigate the correlation between midkine(MK) and VEGF expression with angiogenesis and biological features in recurrent gastric carcinoma.Methods The expression of MK was examined using immunohistochemistry in 9 cases of gastric carcinoma and 9 cases of adjacent normal tissues.Microvessel density(MVD)was determined by VEGF immunohistochemical staining.Results Majority of recurrent gastric carcinoma tissues showed positive reaction to immunostaining,but no specific positivity was detected in adjacent normal tissue.Values of MVD and MK positive group(88.8%,79.75) was higher than those in the negative group(15.0%)(P

Objective To investigate the results of Cierny-Mader type Ⅳ posttraumatic long bone infection treated by en bloc resection combined with induced membrane technique.Methods A retrospective case series analysis was made on 36 patients with posttraumatic long bone infection treated by en bloc resection combined with induced membrane technique from January 2013 to January 2015.There were 30 male and 6 female patients between 21 and 68 years (mean,41 years).Infection control,bone union,function activity and complications were detected after operation.Results After debridement in the first stage,a segmental bone defect of 5.5 cm in length (range,2-10.9 cm) was seen in all patients.Seven patients needed a local flap transfer to cover the wound and five patients had a second debridement.Mean duration of systemic antibiotic use was 2 weeks (range,1-6 weeks) and mean time interval of second-stage bone grafting was 12 weeks after debridement (range,6-36 weeks).Mean follow-up was 29.5 months (range,24-45 months).One patient had recurrence and was cured with radical debridement and a permanent acrylic spacers insertion,with the cure rate of bone infection of 97％ (35/36).All patients achieved bone union with a mean duration of 5.9 months (range,4-8 months),and were able to walk independently.Thirty patients returned to work or pre-operative physical labor.No pain and re-fracture occurred.Six patients had adjacent joint stiffness.Conclusion En bloc resection combined with induced membrane technique is associated with radical debridement,decreased recurrence rate and limb salvage,indicating a simple and effective method for Cierny-Mader type Ⅳ posttraumatic long bone infection.

Objective To identify the differential serum proteins in patients with hepatic injury resulting from coal-burning type of arsenism. Methods Six serum samples were collected from patients with liver injury resulting from coal-burning type of arsenism and healthy subjects(control gruop) in endemic arsenism area. Twodimensional gel electrophoresis(2-DE) was performed to separate serum proteins, after silver staining, the differential expression of proteins were analyzed and then identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS). Results The 2-DE map of serum protein patterns of patients and normal control were established successfully. The results showed that there were an average of (824 ± 31 ) spots and (782 ± 42) spots on 2-DE matching of the patients and control groups and the matching rate was 94.9%(782/824). From these two groups 49 differential protein spots were identified, of which over 3 times the difference in the expression of 30 protein spots were singled out and MALDI-TOF-MS analysis was carried out. Ten proteins were identified. Upregulated expression was observed in alpha-2-macroglobulin, B-cell receptor-associated protein, keratin 1,apolipoprotein A-I, and down-regulated expression was observed in haptoglobin, α2-heremans-schimid-glycoprotein,mitogen-activated protein kinase 4, zinc finger protein 323, ZAP-70 and SP40 in the patient group. Conclusions The well-resolved and reproducible 2-DE serum patterns of patients are established and some differentially expressed proteins are characterized. Whether these proteins of differential expression are serum markers for liver injury resulting from coal-burning type of arsenism need to be further verified.

Objective: To study the preventive effects of heat shock-pretreated rat bone marrow-derived mesenchymal stem cells (MSCs) on the apoptosis of ovarian granulosa cells (GCs) induced by cisplatin. Methods: Rat bone marrow-derived MSCs were isolated, cultured and identified. After heat shock pretreatment for different durations, the apoptotic rates of MSCs were detected to determine the optimal condition of heat shock pretreatment. Cisplatin was added to simulate chemotherapy microenvironment in the ovary. MSCs were divided into normal group (without treatment), heat shock pretreatment group, cisplatin group (without heat shock pretreatment), and heat shock pretreatment+cisplatin group. The apoptotic rate of MSCs was determined. The ovarian GCs were isolated and divided into normal group (without treatment), cisplatin group, MSC prevention group (co-cultured with MSCs before adding cisplatin), and heat shock pretreatment MSC (HS-MSC) prevention group (co-cultured with HS-MSCs before adding cisplatin). The apoptotic rate of GCs was detected. Results: Heat shock pretreatment could reduce the apoptosis of MSCs. After receiving heat shock pretreatment at 42 °C for 1 h, MSCs presented the lowest apoptotic rate. After adding cisplatin, the apoptotic rate of MSCs in heat shock pretreatment+ cisplatin group was significantly lower than that of cisplatin group ([11.94 ± 0.63]% vs [14.30 ± 0.80]%, P<0.05). The apoptotic rate of GCs in HS-MSC prevention+cisplatin group was significantly lower than that of cisplatin group ([39.88±1.65]% vs ([53.81 ±1.89]%, P<0.05). Conclusion: Heat shock pretreatment can alleviate the apoptosis of MSCs during chemotherapy. Heat shock-pretreated MSCs have preventive effects on cisplatin-induced GC apoptosis.

OBJECTIVE:To investigate the differences of the active components of different parts of rhubarb(the head,the body and the end part of the root)from genuine medicinal materials in Gansu province so as to provide theoretical basis for its quality control.METHODS:A comparative study of the fingerprints of different parts of rhubarb extract were conducted by HPLC,in which,the C18 was used as the chromatographic column under room temperature,the mobile phase consisted of menthol-0.1%H3PO4(70∶30)with a flow rate of 1.0ml/min,the column's temperature of room temperature and detection wavel_ ength of 280nm.RESULTS:Significant differences were noted in the HPLC fingerprints of different parts of rhubarb,there were significant differences in the contents of the active components such as emodin,chrysophanol,etc.,the contents decreased progressively from the head of root,the body part to the end part.CONCLUSION:In order to ensure the medical value and the quality of rhubarb medicinal material,different parts of which should be differentiated in the quality control.

OBJECTIVE: To describe our experience in the clinical manifestation and treatment of malignant tumors of the external and middle ear. METHOD: The study reviewed 39 patients between 1994-2011 in our hospital, including 15 pinna tumors, 18 external canal tumors and 6 middle ear tumors. 23 males and 16 females were enrolled in this study. The mean age of patients at the time of surgery was 59. Radiotherapy or radiotherapy and chemotherapy were the only possible treatment in 6 cases. Thirty-three patients were treated surgically, and 9 patients also received radiotherapy after surgery. RESULT: All of the patients had been followed up over 3 years, except for 1 case of external canal and 1 case of middle ear tumor. The 3-year survival of pinna, external canal and middle ear tumors were 86.7%, 82.4% and 60.0% respectively. At the last follow up, the pinna tumors showed that the survival rate was 100% in T1, T2 and Tx stage, and 0% in T4 stage; the external canal tumors showed that the survival rate was 90% in T1 stage, and 66.7% in T2, T3 stage; the middle ear tumors showed that the survival rate was 100% in T1 and T2 stage, 0% in T3 stage. CONCLUSION The T staging system is for an important prognostic factor, and it is important for an early diagnosis and radical surgery to achieve a better therapeutical result.
Ear Auricle ; pathology ; Ear Canal ; pathology ; Ear Neoplasms ; pathology ; Ear, Middle ; pathology ; Female ; Humans ; Male ; Neoplasm Staging ; Retrospective Studies ; Survival Rate

Objective To investigate the effect of silencing E6-associated protein(E6AP)on the level of p53 protein in human papilloma virus(HPV)negative cervical cancer cells(C33A cells).Methods The siRNA sequence silencing E6AP(siE6AP)and silencing control disordered siRNA sequence(siControl)were transfected into C33A cells with the mediation of LipofectamineTM2000 transfection reagent respectively.The silencing effect of siRNA on E6AP and the expression of p53and cleaved-caspase-3 proteins were detected by Western blot.Results The levels of E6AP protein in C33A cells of siE6AP group were significantly lower(t =-4.597,P<0.05),while the levels of p53 and cleaved-caspase-3 proteins were significantly higher than those of siControl group(t = 4.533 and 7.099 respectively,each P<0.05).Conclusion Silencing of E6AP significantly increased the expression of p53 protein in C33A cells,indicating that silencing of E6AP may restore the activity and function of p53 protein in C33A cells.

OBJECTIVETo identify enterovirus 71 (EV71) strains isolated from patients with hand, foot and mouth disease (HFMD) in Guangdong and Fujian provinces from 2000 to 2001 by using phylogenetic analysis.

METHODSAll 25 samples were first tested for enteric viruses by RT-PCR using enterovirus specific primers EV-1 and EV-2, and then were identified for EV71 by RT-PCR using EV71 specific primers 159S and 162A. The amplicons of 485bp segment (part of the VP1 gene) were cloned into pGEM-T and sequenced. A phylogenetic tree was constructed by comparison of the sequences with other 12 EV71 strains isolated from China, Japan, Hungary, and the United States including the prototype BrCr.

RESULTSThe positive rate of EV71 was about 20%. The sequence analysis showed that the new isolate (GZH2000) shared 94%-96% nucleotide identity with three strains isolated in 1998 and 2000, and 91% with a strain isolated in 1987 from Chinese mainland, but shared only 82%-84% homology with EV71 isolates studied abroad.

CONCLUSIONSEV71 is one of the important pathogens of HFMD in south China. The strains isolated from mainland were closely related with most isolates from Taiwan, but different from most EV71 strains reported abroad. The symptoms of EV71 infection in mainland were not as intensive as those described in Taiwan's outbreak.

Adolescent ; Child ; Child, Preschool ; China ; Enterovirus ; genetics ; isolation & purification ; Hand, Foot and Mouth Disease ; virology ; Humans ; Infant ; Phylogeny ; Sequence Analysis ; Sequence Homology, Nucleic Acid