Objective To detect circumsporozoite protein (CSP) in anopheline vectors from south Yunnan and to evaluate ELISA in the detection. \ Methods\ Salivary glands of the anopheline mosquitoes were taken for finding sporozoites by microscopy and part of the glands was used for detecting CSP by ELISA. An. minimus was experimentally infected by blood from vivax malaria patient (with Plasmodium vivax) and examined for sporozoites and CSP. Eight species of anopheline mosquitoes were caught in the field and examined. Monoclonal antibodies to P.falciparum (Pf2A10) and P.vivax (Pv210, Pv247) were used in ELISA for detecting CSP. \ Results\ Sporozoites were found in the salivary glands of 27 out of 36 An. minimus experimentally infected (75\^0%), 29 were ELISA CSP positives (80\^6%), and 26 of the 27 mosquitoes showed Pv210 CSP positive. Among \{1 010\} parous anopheline mosquitoes from the field, 7 were found sporozoite positive (0\^69%), 8 were ELISA CSP positive (0\^79%), and 6 of the 7 mosquitoes showed CSP positive. Of \{4 675\} wild mosquitoes in 8 anopheline species with different ages, 11 were found CSP positive (0\^24%) including An.minimus, An.sinensis and An.maculatus with a positive rate of 0\^20%, 0\^24% and 0\^39% respectively.Among the 11 mosquitoes, 9 were Pv210 positive and 2 were Pf2A10 positive. Conclusion CSP detection by ELISA is a useful method to monitor the malaria transmission capacity of anopheline vectors.

Glutamate decarboxylase (GAD) can catalyze the decarboxylation of glutamate into γ-aminobutyrate (GABA) and is the only enzyme of GABA biosynthesis. Improving GAD activity and thermostability will be helpful for the highly efficient biosynthesis of GABA. According to the Ramachandran plot information of GAD 1407 three-dimensional structure from Lactobacillus brevis CGMCC No. 1306, we identified the unstable site K413 as the mutation target, constructed the mutant GAD by site-directed mutagenesis and measured the thermostability and activity of the wide type and mutant GAD. Mutant K413A led to a remarkably slower inactivation rate, and its half-life at 50 °C reached 105 min which was 2.1-fold higher than the wild type GAD1407. Moreover, mutant K413I exhibited 1.6-fold higher activity in comparison with the wide type GAD1407, although it had little improvement in thermostability of GAD. Ramachandran plot can be considered as a potential approach to increase GAD thermostability and activity.
Glutamate Decarboxylase ; metabolism ; Half-Life ; Industrial Microbiology ; Lactobacillus brevis ; enzymology ; Mutagenesis, Site-Directed ; Mutation ; Temperature

Objective To investigate the distribution patterns of mosquitoes and mosquito-borne viruses in Dehong prefecture,Yunnan province,China. Methods Mosquito samples were collected using the mosquito traps from five counties of Dehong prefecture on July,2007 and 2010. Mosquitoes were cell cultured for viral isolation,and positive isolates were identified using RT-PCR and sequence analysis. Results A total of 43 634 mosquitoes comprised of 29 species representing six genera were collected. Culex tritaeriorhynchus and Anopheles sinensis comprised 78.69% and 14.77% of the total. Six strains of viruses were isolated from the mosquito pools. RT-PCR and phylogenetic analysis revealed three strains from Cx. tritaeriorhynchus,identified as genotypeⅠJapanese encephalitis virus(JEV). One strain was identified from Cx. tritaeriorhynchus,as Getah virus (GETV). Two strains isolated from Cx. tritaeriorhynchus and Anopheles vagus were identified as Culex pipiens pallens Densovirus(CppDNV). Conclusion Cx. tritaeriorhynchus had been the major species of mosquitoes and mainly transmitting vector of mosquito-borne viruses in Dehong prefecture. GenotypeⅠJEV,GETV and CppDNV were the vectors causing transmission of mosquitoe-borne diseases in this area. Data from phylogenetic analysis showed that these newly discovered isolates seemed to have had close relationship with those viruses previously circulating in Yunnan and other provinces of China.

Objcetive: To explore the treatment of long segment defect of tibia by using tensor fascia lata combined with iliac flap or deep circumflex iliac pedicle iliac flap. Methods: From February 2012 to August 2017, The People′s Hospital of Zun Yi City Bo Zhou District treated 16 patients who had long segment defect of tibia.There were 11 males and 5 females, age from 22 to 58 years old, the average age was 42 years old. Iliac flap grafting with tensor fascia lata combined with iliac flap or deep circumflex iliac pedicle was used to treat the defect of long segment of tibia. There were 4 cases with simple tibial defect and 12 cases with skin defect. The longest tibial defect was 5-8 cm. Results: In this study, four patients used iliac flaps with deep circumflex iliac pedicle, the area of flaps ranged from 2.5 cm×5.0 cm to 5.0 cm×10.0 cm, while the area of iliac flaps ranged from 5.0 cm×2.5 cm to 8.0 cm×4.0 cm. Twelve patients used grafting with tensor fascia lata combined with iliac flap, the area of flaps ranged from 5.0 cm×12.0 cm to 12.0 cm×23.0 cm, while the area of iliac flaps ranged from 7.0 cm×2.0 cm to 8.0 cm×4.0 cm. All 16 cases of bone flap were survived, fracture healing, without surgical complications. The average follow-up period was 1.5 years, the flaps had good appearance in 10 cases and was slightly bloated in 6 cases; the ankle had normal motion in 14 cases and had poor dorsal extension in 2 cases. X-ray films showed that the bone flap repaired the bone defects and reached bone healing. Conclusions Vascularized tensor fascia lata combined with iliac flap or deep circumflex iliac pedicle iliac flap grafts increase local blood supply and accelerate the process of fracture healing.

OBJECTIVETo investigate the distribution patterns of mosquito and mosquito-borne viruses in Dehong prefecture, Yunnan province, China.

METHODSMosquito samples were collected using the mosquito traps from five counties of Dehong prefecture on July, 2007 and 2010. Mosquito were cell cultured for viral isolation, and positive isolates were identified using RT-PCR and sequence analysis.

RESULTSA total of 43 634 mosquito comprised of 29 species representing six genera were collected. Culex tritaeniorhynchus and Anopheles sinensis comprised 78.69% and 14.77% of the total. Six strains of viruses were isolated from the mosquito pools. RT-PCR and phylogenetic analysis revealed three strains from Cx. tritaeniorhynchus, identified as genotype I Japanese encephalitis virus (JEV). One strain was identified from Cx. tritaeniorhynchus, as Getah virus (GETV). Two strains isolated from Cx. tritaeniorhynchus and Anopheles vagus were identified as Culex pipiens pallens Densovirus (CppDNV).

CONCLUSIONCx. tritaeniorhynchus had been the major species of mosquito and mainly transmitting vector of mosquito-borne viruses in Dehong prefecture. Genotype I JEV, GETV and CppDNV were the vectors causing transmission of mosquito-borne diseases in this area. Data from phylogenetic analysis showed that these newly discovered isolates seemed to have had close relationship with those viruses previously circulating in Yunnan and other provinces of China.

Alphavirus ; isolation & purification ; Animals ; Arboviruses ; classification ; isolation & purification ; China ; Culicidae ; virology ; Disease Vectors ; classification ; Encephalitis Virus, Japanese ; isolation & purification