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 investigate the effect of exosomes from lung cancer cells on the ability of lung cancer metastasis in-duced by M2 macrophages.Methods:The exosomes secreted by 16HBE,A549 and H522(16HBE exo,A549 exo,H522 exo)were extracted and identified.M0 macrophages co-incubated with 10 μg/ml 16HBE exo,A549 exo,H522 exo,PBS(16HBE exo group,A549 exo group,H522 exo group,PBS group),the expressions of Arginase-1,CD206,CD163,TGF-β,iNOS,and IL-1β in macro-phages in each group were detected by qRT-PCR,the proportion of CD206+CD163+macrophages was detected by flow cytometry,the phosphorylation levels of ERK1/2 and STAT3 in macrophages were detected by Western blot.A549 and H522 cells were co-incubated with macrophages in the above groups(16HBE exo+M0 group,A549 exo+M0 group,H522 exo+M0 group,PBS+M0 group),and the migration and invasive ability of A549 and H522 cells in each group were detected by Transwell chamber method.Results:Compared with PBS group,the expressions of Arginase-1,CD206,CD163 and TGF-β in macrophages in A549 exo group and H522 exo group were significantly up-regulated(P<0.001),and the expressions of iNOS and IL-1β were significantly down-regulated(P<0.001).The proportion of CD206+CD163+macrophages was increased significantly(P<0.001),and the phosphorylation levels of ERK1/2 and STAT3 were increased significantly(P<0.001).There was no significant difference in the above indicators between the 16HBE exo group and the PBS group(P>0.05).The migration ablility and invasion ablility of A549 and H522 cells in A549 exo+M0 group and H522 exo+M0 group were significantly increased(P<0.001).Conclusion:Lung cancer cell exosomes can induce the polarization of M2 macrophages by activating the ERK1/2/STAT3 signaling pathway to promote tumor migration and invasion.

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 investigate the effect of exosomes from lung cancer cells on the ability of lung cancer metastasis in-duced by M2 macrophages.Methods:The exosomes secreted by 16HBE,A549 and H522(16HBE exo,A549 exo,H522 exo)were extracted and identified.M0 macrophages co-incubated with 10 μg/ml 16HBE exo,A549 exo,H522 exo,PBS(16HBE exo group,A549 exo group,H522 exo group,PBS group),the expressions of Arginase-1,CD206,CD163,TGF-β,iNOS,and IL-1β in macro-phages in each group were detected by qRT-PCR,the proportion of CD206+CD163+macrophages was detected by flow cytometry,the phosphorylation levels of ERK1/2 and STAT3 in macrophages were detected by Western blot.A549 and H522 cells were co-incubated with macrophages in the above groups(16HBE exo+M0 group,A549 exo+M0 group,H522 exo+M0 group,PBS+M0 group),and the migration and invasive ability of A549 and H522 cells in each group were detected by Transwell chamber method.Results:Compared with PBS group,the expressions of Arginase-1,CD206,CD163 and TGF-β in macrophages in A549 exo group and H522 exo group were significantly up-regulated(P<0.001),and the expressions of iNOS and IL-1β were significantly down-regulated(P<0.001).The proportion of CD206+CD163+macrophages was increased significantly(P<0.001),and the phosphorylation levels of ERK1/2 and STAT3 were increased significantly(P<0.001).There was no significant difference in the above indicators between the 16HBE exo group and the PBS group(P>0.05).The migration ablility and invasion ablility of A549 and H522 cells in A549 exo+M0 group and H522 exo+M0 group were significantly increased(P<0.001).Conclusion:Lung cancer cell exosomes can induce the polarization of M2 macrophages by activating the ERK1/2/STAT3 signaling pathway to promote tumor migration and invasion.

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.

Purinergic P2Y Receptor Antagonists ; Consensus ; Cardiovascular System ; Cardiology ; Asia

Chronic hepatitis B (CHB) is a global epidemic disease caused by hepatitis B virus that can lead to hepatic failure, even liver cirrhosis and hepatocellular carcinoma. The occurrence and development of CHB are closely related to the changes in the gut microbiota communities. To explore the relationship between the structure of gut microbiota and liver biochemical indicators, 14 CHB patients (the CHB group) and 11 healthy people (the CN group) were randomly enrolled in this study. Our results demonstrate that CHB caused changes in the gut microbiota communities and biochemical indicators, such as alanine transaminase, total bilirubin and gamma glutamyl transferase. Furthermore, CHB induced imbalance of the gut microbiota. Prevotella, Blautia, Ruminococcus, Eubacterium eligens group, Bacteroides uniformis and Ruminococcus sp. 5_1_39BFAA were associated with the critical biochemical indicators and liver injury, suggesting a new approach to CHB treatment.
Bacteroides ; Eubacterium ; Gastrointestinal Microbiome ; Hepatitis B virus ; Hepatitis B, Chronic ; Humans ; Liver Cirrhosis ; Liver Neoplasms

Objective: To investigate the efficacy of enticavir and lamivudine in preventing rituximab-associated hepatitis B virus(HBV) reactivation in patients with B-cell non-Hodgkin lymphoma complicated with resolved hepatitis B during chemotherapy. Methods: This retrospective study included 216 B-cell non-Hodgkin lymphoma patients with complete data from January 2012 to January 2018 treated in 3 hospitals.Of 78 patients with resolved hepatitis B, they were divided into lamivudine prophylactic group(17 cases), entecavir prophylactic group(11 cases) and control group(50 cases). The changes of HBVM, HBV DNA and liver function before or after rituximab combination chemotherapy were analyzed.The incidence of HBV reactivation, liver function injury and chemotherapy delay were compared. Results: Compared to the other 71 patients, 7 cases experienced HBV reactivation in 78 patients with resolved hepatitis B. There were no statistically significant differences between the two groups in patient demographics, pathological pattern, chemotherapy regimen.Six patients in the control group developed HBV reactivation(12%) and 1 patient in lamivudine prophylactic group(5.88%), none had HBV reactivation in enticavir prophylactic group.There was statistically significant difference among three groups(Fisher P=0.016). A total of 7 cases experienced HBV reactivation in 78 patients with resolved hepatitis B respectively, 2 cases in the first chemotherapy period, 2 cases in the third chemotherapy period, 1 case in the fifth chemotherapy period, the last one occurred after the completion of chemotherapy.Four patients had HBsAg reverse seroconversion, occurred in the 1, 3, 5 cycles during and after the completion of chemotherapy.Twenty patients (40.0%) experienced liver function parameters abnormal in the control group, 4 cases(23.5%) in lamivudine prophylaxis group, 2 cases (18.2%) in enticavir prophylaxis group during chemotherapy. Conclusion Antiviral prophylaxic therapy can potentially prevent rituximab associated HBV reactivation in patients with resolved hepatitis B. Entecavir can more reduce the risk of rituximab associated HBV reactivation than lamivudine.

Objective: To determine the changes in peripheral plasma concentrations of interleukin-10 (IL-10), interleukin -12 (IL-12) and interfoeron-γ(IFN-γ) in the patients with chronic hepatitis B virus (HBV) infection and their correlations with HBV infection stage or HBV DNA load of HBV carriers. Methods: Data of 135 patients with chronic HBV infection from March 2016 to March 2017 were collected, the patients included 32 chronic HBV carriers, 61 with chronic hepatitis and 42 with cirrhosis. Forty healthy subjects served as controls. The concentrations of IL-10, IL-12 and IFN-γ were determined using enzyme-linked immunosorbent assay (ELISA). Correlation analysis was performed using the Pearson correlation test, which was performed to analyze the correlation between IL-10, IL-12, IFN-γ and HBV infection stage, HBV DNA load of HBV carriers. Results: Compared with those in healthy controls, plasma IL-10 and IL-12 levels in patients with chronic hepatitis and cirrhosis increased significantly (F=22.06, 15.67, P=0.013, 0.021), plasma IL-10 and IL-12 levels in cirrhosis cases were higher than those in chronic hepatitis (all P<0.001), plasma IL-10 and IL-12 levels in chronic hepatitis were higher than those in chronic HBV carriers (all P<0.001). Plasma IFN-γ level in chronic HBV carriers, chronic hepatitis and cirrhosis were significantly higher than those in healthy controls (F=18.36, P=0.017). There were statistically significant differences in IFN-γ levels among the three groups in the chronic HBV carriers, chronic hepatitis and cirrhosis. IL-10, IL-12 and IFN-γ levels of the low, medium and high HBV DNA load groups were statistically significant (all P<0.05). There was no correlation between IL-10 and HBV DNA. IFN-γ, IL-12 and HBV DNA load were negatively correlated. There was no correlation between IL-10 and IFN-γ (r=0.103, P>0.05), IL-12 and IFN-γ were significantly positively correlated (r=0.687, P<0.05). Conclusions IL-10, IL-12 and IFN-γ may play an important role in the chronic HBV infection.

Objective: To analyze the clinical characteristics of pheochromocytoma crisis (PCC) . Methods: Data of 123 cases of pheochromocytoma and paraganglioma (PPGL) admitted from Apr. 2011 to Feb. 2017 were retrospectively analyzed and they were divided into crisis group and noncrisis group according to the patients with or without haemodynamic instability and end-organ damage. The differences of demographics characteristics, presentations, laboratory tests, imaging findings, perioperative clinical conditions and pathological features were compared between the two groups. Results: ①16 cases were enrolled into crisis group, among whom 5 were misdiagnosed, while 107 cases were enrolled into noncrisis group. ②Compared with noncrisis group, the incidence of headache, palpitation, sweating, the classic triad, other presentations of PPGL, severe hypertension and hypotension were higher, and more patients had paroxysmal hypertension and admitted to our hospital for paroxysmal presentations in crisis group (P<0.05) . ③Leukocyte, fasting blood glucose, liver transaminases, troponin and D-dimmer were higher, while estimated glomerular filtration rate (eGFR) was lower, more tumors located in the left of adrenal in crisis group (P<0.05) . ④ Patients in crisis group had higher plasma free metanephrines (MNs) , larger maximal tumor diameter, higher enhanced CT value in each period, more benign tumors and hemorrhage or necrosis in the tumors, but all the differences were not significant when compared with the noncrisis group. ⑤Patients in crisis group were more likely to undergo elective surgery. However, there was no difference in the preoperative time of α-blockade, type of surgery, intraoperative and postoperative complications, mortality among the two groups. Conclusions PCC is a rare endocrinological emergency with a highly variable manifestations, which commonly presents with typical triad, with higher incidence of hemodynamic instability and end-organ damage. Although biochemical and imaging examinations are relatively effective in the diagnosis of PCC, the misdiagnosis remains inappropriate high. Once the diagnosis is established, clinicians should timely start drug preparation while surgical resection is the key to the treatment of PCC.