Two fragments G1 and G2 of the glycoprotein G gene of bovine respiratory syncytial virus(BRSV) were selected for expression in Escherichia coli based on the analysis of glycoprotein G by DNA Star software.Then the two fragments of glycoprotein G were amplified by PCR with synthesized G gene of BRSV as the template.The amplified fragments G1 and G2 are 570bp and 308bp in length,respectively.The PCR products were cloned into pET30a vector and expressed in soluble form in E.coli after induction of cultured E.coli with IPTG.Both of the recombinant proteins G1 and G2 were purified by immobilized Ni ion affinity chromatography under native conditions.Then the purified proteins were analysed by Western blotting.The results showed that the purified recombinant protein G1 retained good antigenicity and specificity.But the purified recombinant protein G2 didn't possess biological activity.Antibodies against BRSV were detected in suspected bovine serum samples in China by using indirect ELISA and Western blotting with the purified recombinant protein G1.The purified recombinant protein G1 might be used as antigen for establishing serological methods for diagnosis of BRSV infection.And the purified recombinant protein G1 might also be used for preparing polyclonal and monoclonal antibodies for research on biological functions of glycoprotein G of BRSV.

OBJECTIVETo explore the surgical method and clinical curative effect of Ilizarov technique combined with limited surgical treatment for neurotrophic malnutrition equinovarus accompanied with weight-bearing area ulcer.

METHODSFrom July 2004 to December 2011, 21 cases of neurological disorders equinovarus with skin ulcer in weight-bearing area were treated including 14 males and 7 females with an average age of 21.3 years old ranging from 8 to 32 years. Among them,19 cases with talipes equinovarus were on account of spina bifid and 2 cases on account of spinal cord injury of lumbar sacral segment. Nine cases were on the left foot and 12 cases were right foot. The ulcer area in out-below side of the fifth metatarsal bone was in 6 cases, in basement of the fifth metatarsal bone in 5 cases,in lateral of calcaneocuboid joint 2 cases, and in lateral of subtalar joint in 8 cases. Surgical procedure included plantar medial release technique, tendo-chilles lengthenning, and three-joint partial osteotomy. During operation,the skin and soft tissue of ulcer were resected and the incision was sutured when the ulcer was in the lateral of subtalar joint, and ankle Ilizarov external fixation for extension was installed after correcting talipes equinovarus partially. For the legacy skin ulcer in weight-bearing area,the Shenrg-ji cream (Chinese characters) was used after operation for one time per day until the wound healed. Adjusting Ilizarov external fixation for correcting residual deformity until it is satisfaction. During this process weight-bearing walking was keeping.

RESULTSAll cases were followed up from 6 to 52 months with an average of 28 months. All ulcers were healing and no recurrence. The ulcer healing time was 14 to 36 days postoperative (26 days in average). Eighteen deformity feet were corrected satisfactorily, and recovered fully plantar foot, 3 feet appeared mild deformity after 1 year. Ankle joint activity appeared limited in 15 cases and got completely rigidity in 6 cases. The AOFAS increased from preoperative 34.0±7.2 to 86.0±8.5 postoperatively; 8 feet got excellent, 10 good,3 fair.

CONCLUSIONThe clinical effect of Ilizarov technique combined with limited operation and the Sheng-ji cream in correction of the talipes equinovarus with skin ulcer in weigh-bearing area on account of neurotrophic malnutrition is satisfactory, the surgical method is simple and the treating course in security, and serious complications can be avoided.

Adolescent ; Adult ; Child ; Clubfoot ; surgery ; External Fixators ; Female ; Foot Ulcer ; surgery ; Humans ; Ilizarov Technique ; Male

ObjectiveTo investigate the polymorphisms of 124 individual identiifcation SNPs in Chinese Han using the Ion Personal Genome Machine?(PGMTM).Method Samples from 130 unrelated Chinese Han individuals and two families (8 genealogical individuals) were ampliifed using Ion AmpliseqTM Library kit and sequenced on Ion Torrent PGM? platform.Results 14 148 SNPs were detected.A total of 99.992 9% SNPs were correctly called by the HID SNP Genotyper v4.3 plugin, while 0.007 1% wrongly reported and 62 NN calls needed manual correction. The MP ranged from 0.348 0 (rs2831700) to 0.817 3 (rs740910) with the value of 6.898 4 × 10-34 for CMP. The DP ranged from 0.182 7 (rs740910) to 0.652 0 (rs1355366) with the value of 0.999 999 999 999 999 999 999 999 999 999 999 310 2 for CDP, which was larger than that of 22 STR loci. The PE ranged from 0.007 3 (rs1024116) to 0.278 1 (rs1058083) with the value of 0.999 999 616 7 for CPE, which was smaller than that of 22 STR loci. A total of 8 Y-SNP haplo-types were observed from 72 unrelated male samples. No mutation was observed from pedigrees.Conclusion The 124 IISNPs were high polymorphic in Chinese Han and they were ideal markers for human identiifcation. The PGMTM platform has a potential role in forensic science.

Objective To investigate influence of combined use of allopurinol and warfarin on the INR of chronic permanent atrial fibrillation hyperuricemia patients. Methods 80 cases of patients with chronic atrial fibrillation complicated with hyperuricemia, in Sanmen County People's Hospital from February 2016 to February 2017 were randomly divided into the control group (n=40) and the observation group (n=40), the control group was given warfarin plus low purine diet treatment, the observation group was given additional Allopurinol treatment. Changes of INR were compared. Results There was no statistical significance in INR level before treatment between the two groups of patients with permanent atrial fibrillation complicated with chronic hyperuricemia. The INR level of the control group was not changed after treatment, the observation group increased significantly after treatment, and there was statistical significance between the two groups (P<0.05). There were no bleeding events in the control group, there were 6 cases of the observation group with minor bleeding and subcutaneous ecchymosis, including 2 cases with fecal occult blood, 1 cases with hematuria, nasal hemorrhage . The change of liver and kidney function was not obvious. Conclusion In application of allopurinol treatment plus warfarin for patients with chronic persistent atrial fibrillation and hyperuricemia patients, , INR need to be regularly monitored, in order to provide reference for Warfarin dosage adjustment, and ensure clinical safety.

BACKGROUND: The key point of the displaced intra-articular calcaneal fractures is to accurately and effectively reset the intra-articular calcaneal fractures and effectively reduce the incidence of incision-related complications. Currently, the minimally invasive approach has been widely used in the treatment of calcaneal fractures. OBJECTIVE: To compare the clinical outcomes and safety of minimally invasive longitudinal approach and minimally invasive tarsal sinus approach for displaced intra-articular calcaneal fractures. METHODS: A prospective, single center, open-label, randomized, controlled clinical trial is conducted. 200 patients with displaced intra-articular calcaneal fractures will be randomly divided into minimally invasive approach group and tarsal sinus approach group, with 100 cases in each group, and treated with minimally invasive longitudinal approach and tarsal sinus approach repair, respectively. Follow-up time will be 5 weeks, 4, 8, 12, 36 months after surgery. The primary outcome is the excellent and good rate of American Orthopaedic Foot and Ankle Society score at 5 weeks, 4, 8, 12 and 36 months after operation. Secondary outcomes are operative time, CT and X-ray images preoperatively and 5 weeks, 4, 8, 12, 36 months postoperatively at the injury site, and the incidence of adverse reactions 5 weeks, 4, 8, 12, 36 months postoperatively. The trial has been approved by Ethics Committee of Wuxi Third People's Hospital.The study is in line with the Declaration of Helsinki developed by the World Medical Association.Participants will be informed and sign consent for the protocol and procedure. Trial ethics approval [Approved by: Wuxi Third People's Hospital, Approval No. 20140402-2] was conducted in April 2014. The sample and data collection will be from January 2018 to October 2019, and the indicator analysis time and test completion time will be December 2019. The results will be reported in scientific meetings or published in peer-reviewed journals. The trial has registered in China Clinical Trial Register (registration number: ChiCTR-IOR-17013697). DISCUSSION: The trial will compare minimally invasive tarsal sinus approach and minimally invasive longitudinal approach for the treatment of displaced intra-articular calcaneal fractures to clarify the characteristics and clinical application range of the two minimally invasive techniques.

Nine proteins encoded by Mycobacterium tuberculosis RD1 region are important protective antigens that become absent in long passaging of Mycobacterium tuberculosis. They only exist in pathogenic Mycobacteria and are absent in Bacille Calmette-Guerin and environmental Mycobacteria. With good immunogenicities, they may play an important role in the diagnosis and prevention of Mycobacterium tuberculosis. This article reviews recent studies on using RD1-encoded proteins as antigens in the diagnosis of active tuberculosis and tuberculous pleurisy.
Antigens, Bacterial ; isolation & purification ; metabolism ; Humans ; Mycobacterium tuberculosis ; physiology ; Tuberculosis ; diagnosis

OBJECTIVETo study therapeutic effects of limited internal fixation combined with external fixation in the treatment of deformity of distal end of femur, as well as to explore its effect on the function of knee joint.

METHODSFrom June 2004 to June 2009, 38 patients with deformity of distal end of femur were treated with osteotomy at femoral supracondylar and internalfixation combined with hybrid external fixator. Among the patients, 19 patients were male and 19 patients were female, ranging in age from 14 to 58 years, with an average of 26 years. Six patients had genu valgum, 4 patients had varus of the distal end of femur, 2 patients had varus combined with rotation deformity,3 patients had genu recurvation and 23 patients had deformity of genuflex. The function of knee joint was evaluated according to Dimeglio clinical assessment after the treatment.

RESULTSAll the patients were followed up and the during ranged from 6 to 48 months, with a mean period of 23 months. All the deformity obtained satisfactory correction, without complications such as limitation of motion of the knee joint, under correction or over correction,displacement of the section of the osteotomy,internal fixation loosening and fracture. Nineteen patients got an excellent result, 12 good, 5 poor and 2 bad.

CONCLUSIONSupracondylar femoral osteotomy and internal fixation combined with hybrid external fixator is a safe and effective surgery for the treatment of deformity of distal end of femur. This method combines the advantages of internal fixation and external fixation.

Adolescent ; Adult ; External Fixators ; Female ; Femur ; abnormalities ; surgery ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged

Abstract: Objective　To analyze the survival status of HIV/AIDS patients aged above 50 years receiving antiviral therapy (ART) in Shanxi Province from 2011 to 2019, and to provide evidence for further improvement of antiviral therapy. Methods　Basic information and follow-up information of HIV/AIDS patients aged above 50 years who first received HIV/AIDS antiviral therapy in Shanxi Province from 2011 to 2019 were collected. Excel database was established and SPSS23.0 software was used for analysis. Retrospective cohort study was conducted. Cox proportional risk regression model was used to analyze the factors influencing survival time. Results　A total of 1 183 subjects were included, of which 172 died, including 84(48.84%) from other causes, 74(43.02%) AIDS-related death and 14 (8.14%) from accidents, suicides and undetermined deaths. Setting AIDS-related deaths as an outcome event, life table analysis showed that the cumulative survival rates at 1, 3, 5, 7 and 9 years after receiving ART were 96.61%, 93.59%, 90.35%, 87.57% and 83.44%, respectively. Multivariate Cox proportional risk model analysis showed that the risk of death in patients aged 60-<70 years group and over 70 age group was 2.53 times (95%CI: 1.51-4.23) and 3.59 times (95%CI: 1.74-7.40) for patients aged the 50-<60 group , respectively. The risk of death in patients with baseline CD4+T lymphocyte (CD4) counts of ≥200/mm3, 50-<200 /mm3 was 0.22 times (95%CI: 0.12-0.41) and 0.37 times (95%CI: 0.21-0.67) for patients with CD4+T lymphocyte counts of <50/mm3. The risk of death in patients with opportunistic infections at baseline was 1.99 times (95%CI: 1.16-3.39) for patients without baseline opportunistic infections. Conclusions　The survival rate of HIV/AIDS patients aged above 50 who received antiviral therapy (ART) in Shanxi Province from 2011 to 2019 was relatively high. To further improve the quality of antiviral treatment in our province, the strategy of "early detection and early treatment" should be continued and improved in the future, and information collection of specific causes of non-AIDS-related deaths among this population should be further strengthened.

Objective To determine the epidemiological features of hand-foot-and-mouth disease (HFMD) outbreaks and the genetic characteristics of enterovirus type 71 (EV71) isolates from patients in Lianyungang, Jiangsu province in May, 2008. Methods Epidemiological, microbiological, cellular and molecular methods were performed to investigate pathogens and to describe the homogeneity of isolated strains. Results 21 cases were reported in this HFMD outbreak with the attack rate as 20.0%. 3 EV71 virus strains were isolated from 10 stool samples. The nucleotide and amino acid homogeneity of these 3 Jiangsu strain with Anhui Fuyang strains were 97.9%-100.0% and 99.7%-100.0%, respectively. These 4 Jiangsu strains were within genotype C sub-geno group C4 in phylogenetic tree. Data from the follow-up study showed that shedding of EV71 and Coxsackie A 16 virus (CA 16) in the latent period appeared in the outbreak of HFMD. Human beings could be infected by both EV71 virus and CA16 at the same time and could also carry the two viruses. We also discovered that EV71 virus could be expelled out of the human body through stool in the fast week and last for 10 weeks. Conclusion The recently identified EV71 isolates from this HFMD outbreak belonged to sub-geno group CA. Facts as: the release of viruses in the latent period, co-infection or coexisting of two viruses at the same time and super long period of expulsion of toxin exist in EV71 and CA16 did exist.

OBJECTIVETo investigate the correlations of serum interleukin-18 (IL-18) level and IL-18 gene promoter polymorphisms to the development of sepsis in children.

METHODUsing enzyme-linked immunosorbent assay (ELISA), the authors tested the serum IL-18 level in 90 patients with sepsis and 123 normal controls, and their single nucleotide polymorphisms of the promoter region of IL-18 gene at position -607C/A and -137G/C were detected using polymerase chain reaction with sequence specific primers method and sequencing technique.

RESULT(1) The serum IL-18 level in sepsis groups was (196.56 +/- 157.32) pg/ml that was significantly higher than (66.16 +/- 41.63) pg/ml in normal controls (P < 0.01), the more severe the degree of sepsis was, the more significantly higher the serum IL-18 level was. The serum IL-18 level in non serious sepsis group was (152.87 +/- 114.96) pg/ml that was significantly higher than (66.16 +/- 41.63) pg/ml in normal controls, the serum IL-18 level in serious sepsis group was (191.98 +/- 169.72) pg/ml that was significantly higher than that in non serious sepsis group, and the serum IL-18 level in extremely serious sepsis patients was (323.89 +/- 159.35) pg/ml, the difference was highly significant (P = 0.000). The difference was significant among the groups with different severity of sepsis (P < 0.01). There was a negative correlation between PCIS (pediatric critical illness score) of sepsis and the serum IL-18 level (P < 0.01). (2) There were polymorphisms in IL-18 gene promoter of matched healthy children and sepsis in children. The GG genotype frequency (61.8%) of IL-18-137G/C in healthy children was the highest, followed by GC genotype (35.8%) and CC genotype (2.4%) in sequence. The G allele frequency (79.7%) was higher in IL-18-137G/C of healthy children than C allele (20.3%). The GG genotype frequency (71.1%) of IL-18-137G/C in septic children was the highest, the next were GC genotype (26.7%) and CC genotype (2.2%). The G allele frequency (84.4%) was higher in IL-18-137G/C of septic children than C allele (15.6%). The CA genotype frequency (61.0%) of IL-18-607C/A in healthy children was the highest, followed by CC genotype (26.8%) and AA genotype (12.2%). The C allele frequency (57.3%) was higher in IL-18-607C/A of healthy children than A allele (42.7%). The CA genotype frequency (76.7%) of IL-18-607C/A in septic children was the highest, followed by CC genotype (21.1%) and AA genotype (2.2%) in sequence. The C allele frequency (59.4%) was higher in IL-18-607C/A of septic children than A allele (40.6%). (3) The genotype frequency of IL-18-607 CA was 76.7% in sepsis groups that was significantly higher than 61.0% in normal controls, and the genotype frequency of -607 AA was 2.2% in sepsis groups that was significantly lower than 12.2% in normal controls, the difference was significant (P < 0.05). (4) In the order of -137CC, -137GC, -137GG, the serum IL-18 level in normal controls were as follows: (45.67 +/- 28.36) pg/ml, (53.27 +/- 37.91) pg/ml, (76.91 +/- 42.44) pg/ml, and with (140.50 +/- 60.10) pg/ml, (184.42 +/- 157.33) pg/ml, (237.02 +/- 161.76) pg/ml respectively in sepsis groups. In the order of -607AA, -607CA, -607CC, the serum IL-18 level in normal controls were: (48.80 +/- 32.11) pg/ml, (68.41 +/- 42.53) pg/ml, (70.17 +/- 43.87) pg/ml; and with (141.50 +/- 64.35) pg/ml, (151.21 +/- 121.19) pg/ml, (211.16 +/- 163.64) pg/ml respectively in sepsis groups. The difference was not significant among different groups (P > 0.05).

CONCLUSIONThe serum IL-18 level in sepsis groups was significantly higher than that in normal controls, which was related to the severity of sepsis. It was possible that the genotype of -607CA carriers was susceptible to sepsis, which mean that the genotype of -607CA might be susceptible genotype of sepsis. However, the genotype of -607AA might play an oppose role in the risk of sepsis.

Adolescent ; Child ; Child, Preschool ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Infant ; Interleukin-18 ; blood ; genetics ; Male ; Polymorphism, Genetic ; Promoter Regions, Genetic ; Sepsis ; blood ; genetics