Objective:To analyze and predict the biological properties and function of Treponema pallidum OmpH protein by bioinformatics methods, purify the target protein, and prepare polyclonal antibodies. Methods:From January 2024 to February 2025, the research team from the Department of Clinical Laboratory at The First Affiliated Hospital of Hunan Traditional Chinese Medical College (Hunan Province Directly Affiliated Traditional Chinese Medical Hospital) conducted a study employing integrative approaches combining bioinformatics analysis with animal experimentation. During this investigation, the coding sequence of the T. pallidum outer membrane protein H (TpOmpH) was systematically retrieved from the National Center for Biotechnology Information (NCBI) database. And the bioinformatics tools, such as Protparam, Protscale,SignalP 6.0,NetNGlyc-1.0,TMHMM2.0,NetPhos-3.1,SOPMA,AlphaFold3,IEDB,STRING,C-immsim were used to analyze and predict the biological and immunological characteristics of TpOmpH protein. The full length of TpOmpH gene was synthesized and was cloned into the pET28a to construct the recombinant plasmid pET28a-TpOmpH. The the expression of target protein was induced by IPTG and was purified using affinity chromatography. The TpOmpH protein was used to immunize mice and the anti-serum was harvested, then the titer of antibody was detected. Results:TpOmpH is a hydrophobic outer membrane protein with a molecular weight of 19.7 kDa and strong stability. The TpOmpH protein is located outside the cell membrane and contains 11 serine, 4 threonine, and 1 tyrosine phosphorylation site, but no glycosylation sites. The 77.91% of the amino acids in TpOmpH protein are alpha helix, 8.72% are extended strand, 10.47% are random coils, and 2.91% are beta turns. The tertiary structure predicted by AlphaFold3 is in its optimal state. The TpOmpH protein has 4 CTL epitopes, 4 linear epitopes, and 5 spatial epitopes. The TpOmpH protein can interact with Tp92,MutS,SurA,TPANIC_0600 and other proteins which may be involved in Tp invasion. TpOmpH protein can induce an increase in B cell count, antibody content, Th cell count, NK cell count, as well as the expression of various cytokines. High purity TpOmpH protein was obtained through Ni 2+ affinity chromatography, which is consistent with the theoretical molecular weight. TpOmpH protein can induce mice to secrete polyclonal antibodies with antibody titers higher than 1∶10 000. Conclusion:TpOmpH protein is a hydrophobic protein located on the outer membrane of Tp, can induce mice to secrete high titer antibodies, which providing experimental basis for the pathogenesis of Tp and vaccine development.

Objective:To analyze and predict the biological properties and function of Treponema pallidum OmpH protein by bioinformatics methods, purify the target protein, and prepare polyclonal antibodies. Methods:From January 2024 to February 2025, the research team from the Department of Clinical Laboratory at The First Affiliated Hospital of Hunan Traditional Chinese Medical College (Hunan Province Directly Affiliated Traditional Chinese Medical Hospital) conducted a study employing integrative approaches combining bioinformatics analysis with animal experimentation. During this investigation, the coding sequence of the T. pallidum outer membrane protein H (TpOmpH) was systematically retrieved from the National Center for Biotechnology Information (NCBI) database. And the bioinformatics tools, such as Protparam, Protscale,SignalP 6.0,NetNGlyc-1.0,TMHMM2.0,NetPhos-3.1,SOPMA,AlphaFold3,IEDB,STRING,C-immsim were used to analyze and predict the biological and immunological characteristics of TpOmpH protein. The full length of TpOmpH gene was synthesized and was cloned into the pET28a to construct the recombinant plasmid pET28a-TpOmpH. The the expression of target protein was induced by IPTG and was purified using affinity chromatography. The TpOmpH protein was used to immunize mice and the anti-serum was harvested, then the titer of antibody was detected. Results:TpOmpH is a hydrophobic outer membrane protein with a molecular weight of 19.7 kDa and strong stability. The TpOmpH protein is located outside the cell membrane and contains 11 serine, 4 threonine, and 1 tyrosine phosphorylation site, but no glycosylation sites. The 77.91% of the amino acids in TpOmpH protein are alpha helix, 8.72% are extended strand, 10.47% are random coils, and 2.91% are beta turns. The tertiary structure predicted by AlphaFold3 is in its optimal state. The TpOmpH protein has 4 CTL epitopes, 4 linear epitopes, and 5 spatial epitopes. The TpOmpH protein can interact with Tp92,MutS,SurA,TPANIC_0600 and other proteins which may be involved in Tp invasion. TpOmpH protein can induce an increase in B cell count, antibody content, Th cell count, NK cell count, as well as the expression of various cytokines. High purity TpOmpH protein was obtained through Ni 2+ affinity chromatography, which is consistent with the theoretical molecular weight. TpOmpH protein can induce mice to secrete polyclonal antibodies with antibody titers higher than 1∶10 000. Conclusion:TpOmpH protein is a hydrophobic protein located on the outer membrane of Tp, can induce mice to secrete high titer antibodies, which providing experimental basis for the pathogenesis of Tp and vaccine development.

Objective To observe the effects of Yiqi Jiedu Compound on TCM syndrome score and immune regulation of Th17/Treg cells in peripheral blood of patients with myasthenia gravis.Methods 63 patients with myasthenia gravis were randomly divided into the control group(n=31)and the treatment group(n=32).The control group was given the basic treatment of pyridostigmine bromide and glucocorticoid and so on,and the treatment group was given Yiqi Jiedu Compound based on the control group.The two groups were treated for 12 weeks.Clinical efficacy was observed,TCM syndrome scores were statistically analyzed,and the expression levels were detected,which Th17/Treg cells,Foxp3/RORγ t gene and IL-6 and IL-17 cytokines in peripheral blood of patients in both groups before and after treatment.Results After 12 weeks of treatment,compared with the control group,the TCM syndrome score of patients in the treatment group was significantly decreased(P<0.05),and the expression of Foxp3 and Treg genes in peripheral blood was promoted(P<0.05),and the expression of Th17 and Treg cells and IL-6 and IL-17 cytokines in peripheral blood was decreased(P<0.05).Conclusion Yiqi Jiedu Compound may regulate the immune balance of Th17/Treg cells by affecting the differentiation of Th17 and Treg cells in peripheral blood and the expression levels of transcription factors RORγ t and Foxp3 genes and related cytokines,thus achieving the effect of myasthenia gravis treatment.

Objective To study the effect of Yiqi Jiedu compound on autoimmune regulator(Aire)and transcription factor retinoic acid-related orphan receptor(ROR-γt)/Effects of forkhead wing helix transcription factor 3(Foxp3)-related inflammatory factor expression.Methods The EAMG model was replicated by immunization-induced injection of mouse-derived AchR-α subunit 97-116 peptide,including normal group,model group,positive control group(prednisone),and Yiqi Jiedu compound high-dose,medium-dose and low-dose groups.Group of 10,the behaviors and Lennon scores of the rats in each group were observed.ELISA method was used to detect serum transforming growth factor-β1(TGF-β1)and interleukin-6(IL-6)levels.Western blot was used to detect the thymus tissue of EAMG rats.The expression of Aire and Foxp3,and the expression of RORγt in the thymus tissue of rats in each group was detected by RT-PCR.Results Compared with before treatment,the Lennon grading score of each drug group was significantly decreased;compared with the prednisone group,there was no significant difference in the Lennon score of each dose group of Yiqi Jiedu compound(P>0.05).RT-PCR detection results showed that compared with the normal group,the relative expression of RORγt mRNA in the thymus of the model group was significantly increased(P<0.01).Western blot detection results showed that the expressions of Aire and Foxp3 in the model group were significantly lower than those in the normal group(P<0.05);The levels of Aire and Foxp3 proteins in thymus of the rats in each drug group were significantly higher than those of model group(P<0.01).ELISA results showed that serum IL-6 content in model group was significantly higher than that in normal group(P<0.01),while TGF-β1 content was significantly lower than that in normal group(P<0.01).After intragastric administration,the content of IL-6 in all drug groups was significantly decreased,and the content of TGF-β1 was significantly increased(P<0.01).Conclusion Yiqi Jiedu compound can improve the symptoms of muscle weakness in EAMG rats,and its mechanism may be through up-regulating the expression of Aire protein,regulating immune inflammatory response,affecting the differentiation of Th17/Treg,thereby regulating the immune balance of ROR-γt/Foxp3.The role of autoimmune response may be one of the mechanisms of its treatment of MG.

OBJECTIVE：To study the effects of chrysophanol on the activa tion of microglia and the expression of inflammatory factors in cerebral ischemia model rats. METHODS ：SD rats were randomly divided into sham operation group ， model group and chrysophanol high ，medium，low dose groups [7.88，3.94，1.97 mg/（kg·d）]，with 20 rats in each group （the number was complemented in cases of death or unsuccessful modeling during modeling process ）. Except for sham operation group ， middle cerebral artery occlusion model was established in other groups by improved thread method. After 2 hours of ischemia ， sham operation group and model group were intraperitoneally injected with 1 mL normal saline ，and each administration group was intraperitoneally injected with 1 mL corresponding drug ，once a day ，for 7 consecutive days. After last medication ，the score of neurological impairment was recorded ；cerebral infarction of rats was observed by TTC staining ，and the percentage of cerebral infarction area was calculated. The expression of Iba- 1 positive cells in ischemic penumbra of rats was observed by immunofluorescence staining. The expression of Notch- 1，TNF-α and ICAM-1 in the ischemic penumbra of rats were detected by Western blotting assay. RESULTS ：In sham operation group ，there was no infarction area in the brain tissue ，and the Iba- 1 positive cells in the ischemic penumbra were few and branched. Compared with sham operation group ，the infarction area of cerebral tissue in rats was obvious in model group ； the 052）number of Iba- 1 positive cells in ischemic penumbra were 〔ZQ2017003〕） increased significantly ，and they were in amoeba or round shape；the neurological impairment score ，the percentage of cerebral infarction area ， relative expression of Notch- 1， TNF-α and ICAM-1 protein in ischemic penumbra were increased significantly （P＜0.05）. Compared with m odel rats ，the infarction area of cerebral tissue in each dose group of chrysophanol was reduced to different extent ；the number of Iba- 1 positive cells in ischemic penumbra was decreased ；neurological impairment score ，the percentage of cerebral infarction area ，relative expression of Notch- 1，TNF-α and ICAM-1 protein were significantly decreased （P＜0.05 or P＜0.01）. CONCLUSIONS ：Chrysophanol has a certain protective effect on the brain tissue of cerebral ischemia model rats ，and can relieve the nerve injury. Its mechanism may be associated with inhibiting the activation of microglia and expression of inflammatory factors mediated by Notch pathway.

BACKGROUND: There is often a physical and psychosocial effect in burn injury, and the degree of mental stress reaction is related to social support, individual experience and personality characteristics.OBJECTIVE: To investigate the mental stress reaction, the degree of social support and the Minnesota Multiphasic Personality Inventory (MMPI)personality characteristics in acute burn patients, and to analysis the influence factor of mental stress reaction.DESIGN: Cross-sectional study.SETTING: West China Mental Health Center of Sichuan University, Burn Department of Mianyang Municipal Central Hospital, and Burn Department of General Hospital of Panzhihua Iron and Steel Company.PARTICIPANTS: From April 2003 to April 2005, totally 104 mild, moderate or severe burn patients, including 92 males and 12 females, aged 12-60 years, selected from Mianyang Central Hospital and Department of burn of General Hospital of Panzhihua Iron & Steel Co., Ltd took part in the study.METHODS: The emotional problems and personality characteristics were measured with the Hamilton Rating Scale for Depression (HAMD), the Hamilton Rating Scale for Anxiety (HAMA), the Minnesote Multiphasic Personality Inventory (MMPI), the Social Support Rating Scale (SS) as well as general clinical data,by the rating scales 3 day after burn. The patients were divided into anxiety-depression group (n=51) and normal emotion group (n=53) according to their anxiety-depression scores (anxious score＞ 7,depressive score＞ 8), those influencing factor included burning total area,age, sex and society support, Stepwise Regression was introduced to get the main factors that influenced mental stress reaction.MAIN OUTCOME MEASURES: ① Scores of each scale and comparison of general clinical data in each group; ② stepwise regression analysis of depression and anxiety of burn patients.RESULTS: Data of Totally 104 patients was entered the final analysis. ①The scores were 10.4±2.46 and 16.8±5.94 of anxious and depressive patients during acute burn period, which were higher than those in normal group (4.29±2.26, 5.48±2.06, P=0.01). ② The main influential factor for depression were burning area and social supporting availability, and those for anxiety were age, burning area, social supporting availability and subjective social supporting degree. ③ MMPI test results show that T scores in anxiety-depression group were below 60 points, however T-scores were above 60 points in normal group including cheating points, hypomania,schizophrenia, psychasthenia and paranoid ideation CONCLUSION: Burn is a mental crisis to burn patients, which can lead to abnormal emotional reaction and personality change. The depressive and anxious score are positively related with burning area, and are negatively correlated with society supporting availability. The good social support system is benefit to over mental crisis period.

Objective To explore the differences of intelligence ，ability of social adaptation and behavior problems between patients of benign epilepsy childhood and those of normal controls .Methods Sixty children of benign epilepsy were investigated. Results The intelligence of children showed no significant differences between two groups ( P＞0.05), while the rate of ability of social adaptation among the children of benign epilepsy patients was less than that of nomal controls （P<0.01) . There were no significant differences in the total rate of behavior problems between two groups except that some factors were different . Conclusions We should create methods to enhance the ability of social adaptation in order to avoid the abnormal behavior when the children patients of benign epilepsy are treated by drugs.