Objective To develop a rapid molecular biological method for detection of the asymptomatic infection of Leish?mania. Methods Two pairs of primers named RV1?RV2 and K13A?K13B were selected to be the fast diagnosis primers since they were designed according to the conserved region of Leishmania kinetoplast DNA(kDNA)minicircles. The PCR amplifica?tion products of Leishmania donovani promastigote from Shandong Province were sequenced to compare their conservatism. The method was applied to detect 105 venous blood samples from healthy home canine and 7 venous blood samples from home canine suffered from Kala?azar in Heishui County of Sichuan Province,and 75 venous blood samples from susceptible population(no leishmaniasis symptoms)and 7 venous blood samples from patients in Xinjiang Kashi area in order to verify the feasibility and accuracy of the method. Results The size of PCR products was consistent with the expected fragments with high conservative among Leishmania species. The positive rates of 105 home canine samples and 75 susceptible population samples were 37.14%(39/105)and 82.67%(62/75)rspectively,and the positive rates of home canine suffered from Kala?azar and patients were all 100%(7/7). Conclusion This rapid diagnosis method is suitable for detection of asymptomatic infection of Leishmania in Kala?azar endemic areas of China with high sensitive and specific,thus it has bright perspective to be used.

Objective To establish the primary cat intestinal epithelial cells(IECs)culture methods and construct the cD-NA library for the following yeast two-hybrid experiment,so as to screen the virulence interaction factors among the final host. Methods The primary cat IECs were cultured by the tissue cultivation and combined digestion with collagenase XI and dispase I separately. Then the cat IECs cultured was identified with the morphological observation and cyto-keratin detection ,by using goat anti-cyto-keratin monoclonal antibodies. The mRNA of cat IECs was isolated and used as the template to synthesize the first strand cDNA by SMARTTM technology,and then the double-strand cDNAs were acquired by LD-PCR,which were subsequently cloned into the plasmid PGADT7-Rec to construct yeast two-hybrid cDNA library in the yeast strain Y187 by homologous recom-bination. Matchmaker?Insert Check PCR was used to detect the size distribution of cDNA fragments after the capacity calcula-tion of the cDNA library. Results The comparison of the two cultivation methods indicated that the combined digestion of colla-genase XI and dispase I was more effective than the tissue cultivation. The cat IECs system of continuous culture was established and the cat IECs with high purity were harvested for constructing the yeast two-hybrid cDNA library. The library contained 1.1× 106 independent clones. The titer was 2.8 × 109 cfu/ml. The size of inserted fragments was among 0.5-2.0 kb. Conclusion The yeast two-hybrid cDNA library of cat IECs meets the requirements of further screen research,and this study lays the foundation of screening the Toxoplasma gondii virulence interaction factors among the cDNA libraries of its final hosts.

Objective To explore the biological function of rhoptry protein 38(ROP38)of Toxoplasma gondii,and to iden?tify the reactogenicity of the recombinant protein(rROP38). Methods The ROP38 was amplified by RT?PCR from T. gondii RH strain,and was cloned into prokaryotic expression vector pET?28a(+). The recombinant plasmid was transformed into E. co?li BL21(DE3)competent cells. Then the rROP38 was analyzed by SDS?PAGE and identified by Western blot. Results SDS?PAGE showed that rROP38 was efficient expression with a molecular weight of about 43 kD. Western blot showed that rROP38 reacted with antibody of His tag or human positive antibody,which indicated that ROP38 had good reactogenicity and could be a serological diagnostic antigen. Conclusion The study successfully obtains the rROP38 of T. gondii with good reactogenicity.

Objective:To evaluate internal fixation via only the modified Stoppa approach in the treatment of central hip dislocation complicated with fracture of the posterior acetabular wall.Methods:A retrospective study was conducted in the 13 patients with central hip dislocation and fracture of the posterior acetabular wall who had been treated at Department of Orthopedic Trauma, Nanfang Hospital between February 2015 and February 2018. They were 10 men and 3 women, aged from 31 to 65 years (average, 46.7 years). All patients were treated with internal fixation via only the modified Stoppa approach. The reduction of double-column and posterior wall fractures was evaluated according to the X-ray Matta scoring system, as well as to the Wiberg central-edge (CE) angles between the vertical line of the center point of the femoral head and the lateral edge of the acetabulum and acetabular tolerance on the normal and affected sides immediately after operation; the hip function was evaluated by the modified Merle d'Aubigne and Postel scoring system at 12 months after operation.Results:All patients were followed up for 16 to 52 months (average, 25.6 months). In all of them, reduction and fixation of central hip dislocation and acetabular fracture was completed successfully, and indirect reduction of posterior wall fracture and acetabular tolerance were satisfactory. Operation time ranged from 130 to 270 min, averaging 155.5 min; intraoperative blood loss from 600 to 5,600 mL, averaging 1,150.5 mL; intraoperative infusion of concentrated red blood cells from 2 to 12 U, averaging 6 U. By the X-ray Matta scoring system immediately after operation, anatomical reduction was achieved in 4 posterior wall fractures and satisfactory reduction in 9 ones. There was no significant difference between the normal and affected sides in the CE angle (43.53°±3.46° for the affected side versus 43.19°±3.28° for the normal side) or in the acetabular tolerance (76.56%±15.50% for the affected side versus 75.32%±16.24% for the normal side) ( P>0.05). The modified Merle d'Aubigne and Postel scores at 12 months after operation ranged from 12 to 18 points, averaging 16.5 points; the hip function was assessed as excellent in 9 cases, as good in 3 and as fair in one. By the last follow-up, none of the 13 patients lost fracture reduction, and their internal fixation was firm with no loosening or breakage. Conclusion:In the treatment of central hip dislocation complicated with fracture of the posterior acetabular wall, internal fixation via only the modified Stoppa approach can lead to satisfactory fracture reduction, firm fixation, good hip joint tolerance, and fine clinical efficacy.

Objective:To analyze the clinical efficacy of bone transport with unilateral external fixation in the treatment of massive tibial bone defects.Methods:A retrospective study was conducted to review the 21 patients with massive tibial bone defects who had been treated by bone transport with unilateral external fixation from February 2017 to January 2022 at Department of Trauma Orthopedics, Ganzhou People's Hospital. There were 14 males and 7 females with a mean age of (46.3 ± 11.3) years. Causes for bone defects: trauma ( n=5), resection of bone non-union ( n=9), resection of infected bone ( n=6) and resection of bone tumor ( n=1). The mean bone defect length was (8.3 ± 1.7) cm. Bone transport started from 10 to 12 days after operation, with a speed of 1 mm/d, and was completed in 4 times. X-ray films were reviewed every 2 weeks. The bone union time, external fixation time (EFT), external fixation index (EFI), docking site situation and complications were recorded. The clinical efficacy was assessed by Paley score. Results:All patients were followed-up for a mean time of (13.5 ± 5.5) months. The mineralization of regenerated bone was good. The bone union time was (9.6 ± 2.2) months, the EFT (10.3 ± 4.0) months, and the EFI (1.3 ± 0.4) months/cm. All docking sites got united. The docking sites were cleaned in 14 patients, of whom simple compression with external fixation was performed in 5 and bone grafts at the docking sites in 9. Postoperative nail tract infection was observed in 6 cases, tibial alignment deviation in 1 case, foot drop deformity in 5 cases, horseshoe varus foot deformity in 1 case, toe flexion deformity in 3 cases, and refracture after removing the external fixation in 1 case. By the Paley score, the bony outcomes were rated as excellent in 16 and as good in 5 cases. The functional outcomes were excellent in 10, good in 7, and acceptable in 4.Conclusion:Bone transport with unilateral external fixation is an effective treatment for massive tibial bone defects, showing advantages of easy operation and convenient carry.

Trauma is the leading cause of death for people under 40 years old in the world. At present, the rescue and treatment system of trauma patients in China is not yet well established, and the mortality of trauma patients is higher than those in the developed countries. Improving the treatment system is the key to reducing the trauma mortality. In order to innovate the service mode of trauma first aid, further promote the establishment of regional trauma first aid system, improve the ability of trauma treatment, reduce the mortality and disability rate of trauma patients in Jiangxi Province, recently Health Commission of Jiangxi Province and the First Affiliated Hospital of Nanchang University have reached a consensus on the establishment of Jiangxi trauma first aid center. In order to provide reference for the construction of trauma treatment system, the author analyzes the following aspects including functional positioning, basic requirements, organization management, and evaluation of core indicators.