Objective To high express and purify toxoplasma gondii antigen GRA6 in E.coli which can be used to develop the genetic engineering diagnostic reagents.Methods The recombinant plasmid of pGEX-GRA6 was transformed to a bacterium BL21-Codon Plus(DE3)-RP and the recom- binant product was expressed under the inducement of isopropyl-beta-D-thiogalactosidase(IPTG). Cells were lysed by multiple rounds of sonication.The expression product was analyzed by using SDS- PAGE.Furthermore,it was purified by sedimentation of ammonium sulphate,desalting using Sephe- dax GS0 and affinity chromatography on glutathione-sepharose system.The immunogenicity of recom binant antigen purified was tested with Western blot.Results GRA6 was highly expressed in E.coli as fusion protein consisting of glutathione S-transferase and GRA6(GST-GRA6).The solubility anal- ysis of expression product indicated that this recombinant protein could be expressed in both superna- tant and inclusion bodies.The soluble protein of GRA6 in supernatant yield the final preparation at greater than 90% after purification.The recombinant protein purified could be recognized by human toxoplasmosis-infective sera with Western blotting analysis.Conclusions The soluble protein of GRA6 was highly expressed in E.coli and the recombinant antigen could be recognized by human tox- oplasmosis-infective serum after purification.The recombinant antigen can be used for devoloping kit to diagnose the acute and chronic infection of toxoplasma gondii.

Objective To evaluate the relationship between varicocele (VC) and the prostativenouplexuby coloDoppleflow imaging(CDFI) and to explore the etiology of varicocele .MethodThe innediameterand the hemorrheologiparameterof spermativein and prostativenouplexuwere observed in 135 patientwith lefvaricocele(lefVgroup) ,51 patientwith bilat-eral V(bilateral Vgroup) and the control group(100 cases) by CDFI .The diameteof the prostativenouplexus(PVD) ,peak velocity of reflux flow (RFV) in the Valsalvtesand the peak velocity of antegrade flow (AFV) aresin 3 groupwere statistical-ly analyzed .ResultPVD and RFV in the bilateral Vgroup were greatethan those in the lefVgroup and the control group (P＜0 .01) .PVD and RFV in the lefVgroup had no statistical differencecompared with the control group (P＞0 .05) .AFV had no statistical difference among 3 group(P>0 .05) .PVD ,RFV and AFV in 30 caseof Vhad no statistical differencebe-tween before and afteoperation (P>0 .05) .Conclusion Bilateral Vmay be accompanied with potential systematic vascular abnormalities.

To investigate the effect of Dachengqi decoction on NF-kappaB p65 expression in lung of rats with partial intestinal obstruction and the underlying mechanism, 30 SD rats were randomly divided into three groups: sham-operation group, model group and Dachengqi decoction treatment group (Dachengqi group), with 10 animals in each group. The models were made by partially ligating their large intestines outside the body. The pathological changes were analyzed by HE staining. The expression of NF-kappaB p65 in rats lung were measured by using real-time polymerase chain reaction and immunohistochemistry respectively. Moreover, the expression of caveolin-1 in rats lung was also measured to. Increased edema, interstitial thickening, hemorrhage, and infiltration of inflammatory cells were found in the model group. In contrast, this change was significantly reduced in Dachengqi group as compared with model group. In addition, the up-regulated caveolin-1 and NF-kappaB p65 were also suppressed by Dachengqi decoction in lung of rats with partial intestinal obstruction. We are led to concluded that the caveolin-1-NF-kappaB pathway plays an important role in the development of lung injury of rats with partial intestinal obstruction and Dachengqi decoction could down-regulate the expression of caveolin-1 and NF-kappaB p65 in lung of rats with partial intestinal obstruction.

Objective To investigate the detection methods of atypical bcr-abl rearrangement with b3a3 fusion transcript,and to describe the characteristics of this fusion gene.Methods Karyotype analysis,FISH and RT-PCR were applied to detect the break point of bcr-abl fusion gene in a patient who was diagnosed as acute lymphoblastic leukemia.Results The karyotype of the patient was expressed as 45,XY,-7,t(9;22)(q34;q1 1).The translocation event in chromosome 9 and 22 could be successfully detected by FISH,and a rare bcr-abl rearrangement with b3a3 fusion transcript was detected by RT-PCR with specific primers.Conclusions The rare e14a3 (b3a3) fusion of bcr-abl gene is present in this patient.Clinical laboratories using commercial kits that do not cover such rare fusions are likely to generate false result,thereby declaring combination of various methods to detect fusion genes is necessary.More studies are needed to explore the function and significance of rare bcr-abl fusion genes.

Objective To investigate the distribution and drug resistance of pathogenic bacteria in patients with preg-nancy combined with urinary tract infection. Methods Mid-stream urine culture and drug susceptibility analysis were conducted on 552 patients with pregnancy combined with urinary tract infection. Results The top 3 pathogenic bacteria for pregnancy combined with urinary tract infection were escherichia coli, enterococcus faecium and acinetobacter bau-mannii. Escherichia coli remained low resistance rate to piperacillin-tazobactam (1.99%), imipenem (1.49%), amikacin (4.47%), cefperazone-sulbactam (2.98%) and macrodantin (3.98%), but high resistance rate to ciprofloxacin (73.13%) and levofloxacin (67.16%). Conclusion The spectrum and drug resistance rate of pathogenic bacteria in pregnancy combined with urinary tract infection are changing. The pathogenic bacteria in urine and their drug resistance should be closely monitored to provide reference for clinical drug use.

To investigate the effect of Dachengqi decoction on NF-κB p65 expression in lung of rats with partial intestinal obstruction and the underlying mechanism,30 SD rats were randomly divided into three groups: sham-operation group,model group and Dachengqi decoction treatment group (Dachengqi group),with 10 animals in each group.The models were made by partially ligating their large intestines outside the body.The pathological changes were analyzed by HE staining.The expression of NF-κB p65 in rats lung were measured by using real-time polymerase chain reaction and immunohistochemistry respectively.Moreover,the expression of caveolin-I in rats lung was also measured to.Increased edema,interstitial thickening,hemorrhage,and infiltration of inflammatory cells were found in the model group.In contrast,this change was significantly reduced in Dachengqi group as compared with model group.In addition,the up-regulated caveolin-1 and NF-κB p65 were also suppressed by Dachengqi decoction in lung of rats with partial intestinal obstruction.We are led to concluded that the caveolin-1-NF-κB pathway plays an important role in the development of lung injury of rats with partial intestinal obstruction and Dachengqi decoction could down-regulate the expression of caveolin-1 and NF-κB p65 in lung of rats with partial intestinal obstruction.

Objective:To evaluate skin irritation,acute toxicity,and allergy of medical ozone oil for clinical application.Methods:In contrast to their left and right side irritation,one or more skin irritation tests were performed on the intact and damaged skins of guinea pigs.With the maximum concentration,acute skin toxicity test was applied on the intact and damaged skins of rats.Active cutaneous anaphylaxis was applied to the guinea pigs.Results:High concentration (ozone consumption:150 g/L) of medical ozone oil showed a slight irritation on the broken skin of guinea pigs,while low concentrations did not show skin irritation.Medical ozone oil had no obvious acute toxicity to rats.The medical ozone oil and base oil showed mildallergy for the skin of guinea pig.Conclusion:The irritation of medical ozone oil is related to its concentration.With appropriateconcentration and duration of treatment,medical ozone oil is safe.

【Objective】 To map a comprehensive description of microbiome presented in healthy blood donations in Liangshan Yi Autonomous Prefecture, and detect the potential pathogens and its prevalence. 【Methods】 A total of 1 299 blood samples were randomly selected from healthy blood donors in Liangshan Prefecture. Total nucleic acids were extracted and sequenced by high-throughput sequencing, and the microbiome was determined by metagenomics analysis. Quantitative real-time PCR (qPCR) and ELISA assays were used to detect antibodies against HSV DNA and HSA in each sample. The prevalence of HSV in healthy blood donors was compared in terms of gender, age, occupation, education level, and frequency of donation. 【Results】 3.49GB data were obtained from DNA pool through high-throughput sequencing. After filtering the data of human genome, the DNA sequences annotated as bacteria, parasites / fungi and viruses were 213 057, 10 623 and 15 reads, respectively. A total of 2.79GB data were obtained from cDNA pool, after filtering the data of human genome, the fragments annotated as bacteria, parasites / fungi and viruses were 4 105 600, 18 446 and 0 reads, respectively. The prevalence of IgG and / or IgM antibodies against herpes simplex virus type 1 and 2 were 8.62% (112 / 1 299) and 18.32% (238/1 299), and that of nucleic acid was 0.77 ‰ (1/1 299). 【Conclusion】 The microbiome of healthy blood donors in Liangshan Prefecture and the potential pathogens were identified in this study. Regional specificity of HSV infections emerged in Liangshan Prefecture. Considering the collaboration between HSV-2 and HIV infection, epidemiological investigation of HSV-2 infection should be conducted preferentially among different populations in Liangshan Prefecture and other HIV high prevalence areas in order to benefit the prevention and control of HIV.

【Objective】 To determine the ELISA kit for screening convalescence plasma with high potency of SARS-CoV-2 IgG by comparing and analyzing the plasma detection results of convalescent plasma collected in different periods via ELISA kits from two manufacturers and the results of mixed plasma with different potency via pseudovirus neutralization experiments. 【Methods】 Two ELISA kits from different manufacturers(named A, B) were used to detect the plasma of 269 convalescent patients collected from Feb.2020~Jan.2022. The correlation and concordance rate of the two results were analyzed to determine the kit preliminarily. According to the titers of diluted series of standard of the preliminary selected kit, 5 mixed plasma samples (G4-G128) with different potency were prepared. The correlation of ELISA IgG results of product A/B, as well as the pseudovirus neutralization test of the original strain, Omicron mutant BA.1 and BA.2 strains were analyzed. Combined with the outside-well dilution mode of the strongly positive samples, the kit for high potency of SARS-CoV-2 IgG screening was determined. 【Results】 When the internal control reference B2 was used as the standard, the detection sensitivity of product A and B was 1∶32 vs 1∶8; the detection sensitivity of product A was 4 times that of product B. The correlation Pearson r between the results given by two kits was 0.944 1(P<0.000 1). Product B with low sensitivity was primarily selected as an alternative kit. The ELISA IgG results of samples from mixed plasma showed that the order of correlation r between product A and B was 0.988. The correlation r between product A and neutralization antibody potency of the three viruses was original strain (0.978)>BA.2(0.970)>BA.1(0.799); the order of correlation r between ELISA IgG results of product B and neutralization antibody potency of the three viruses was original strain(0.994)>BA.2(0.968)>BA.1(0.804). If twice-diluted B2 was taken as the excellent standard, 55.4% of product B met the criterion, while 47.2% of product A met.For positive plasma with high IgG potency, the product B kit required a lower dilution of the sample, which was more convenient to operate. 【Conclusion】 Both of the ELISA IgG kit from product A and B can be used to screen IgG antibodies of SARS-CoV-2, while product B is more suitable for screening positive plasma with high IgG potency.

Objective: To compare the difference of efficacy between traditional Hyper-CVAD/MA regimen and the adolescents inspired chemotherapy regimen, CH ALL-01, in treatment of adult Philadelphia chromosome-positive acute lymphoblastic leukemia (Ph⁺ ALL) . Methods: In this study we retrospectively analyzed 158 Ph⁺ ALL patients receiving Hyper-CVAD/MA regimen (n=63) or CHALL-01 regimen (n=95) in our center and Changzheng hospital from January 2007 to December 2017, excluding patients with chronic myeloid leukemia in blast crisis. Tyrosine kinase inhibitor (TKI) was administered during induction and consolidation chemotherapy. Patients who underwent hematopoietic stem cell transplantation received TKI as maintenance therapy. Results: Of them, 91.1% (144/158) patients achieved complete remission (CR) after 1-2 courses of induction. CR rate was 90.5% (57/63) for patients in Hyper-CVAD/MA group and 91.6% (87/95) for patients in CHALL-01 group. There was no difference in CR rates between the two groups (χ²=0.057, P=0.811) . The last follow-up was June 2018. A cohort of 134 CR patients could be used for further analysis, among them, 53 patients received Hyper-CVAD/MA regimen and other 81 patients received CHALL-01 regimen. The molecular remission rates were significantly higher in CHALL-01 group (complete molecular response: 44.4%vs 22.6%; major molecular response: 9.9% vs 18.9%) (χ²=7.216, P=0.027) . For the patients in Hyper-CVAD/MA group, the 4-year overall survival (OS) was 44.81% (95%CI: 30.80%-57.86%) and the 4-year disease free survival (DFS) was 37.95% (95%CI: 24.87%-50.93%) . For patients received CHALL-01 regimen, the 4-year OS was 55.63% (95%CI: 39.07%-69.36%) (P=0.037) and 4 year DFS was 49.06% (95%CI: 34.24%-62.29%) (P=0.015) , while there was no significant difference in 4 year cumulative incidence of relapse (CIR) (P=0.328) or cumulative incidence of nonrelapse mortality (CI-NRM) (P=0.138) . The rate of pulmonary infection was lower in patients received CHALL-01 regimen compared with patients received Hyper-CVAD regimen (43.4% vs 67.9%, χ²=7.908, P=0.005) . Conclusions Outcome with CHALL-01 regimen appeared better than that with the Hyper-CVAD/MA regimen in Ph⁺ ALL, which has lower incidence of pulmonary infection, higher molecular remission rate and better OS and DFS.