Objective To investigate the effects of Echinococcus multilocularis infection on levels of memory T (Tm) cells and their subsets in lymph nodes of mice at different stages of infection, so as to provide new insights into immunotherapy for alveolarechinococcosis. MethodsTwenty-four C57BL/6J mice aged 6 to 9 weeks were randomly divided into the infection group and the control group, of 12 mice in each group. Mice in the infection group were administered with 3 000 E. multilocularis protoscoleces via portal venous injection, while animals in the control group were administered with an equal volume of physiological saline. Three mice from each group were sacrificed 4, 12 weeks and 24 weeks post-infection, and lymph nodes were sampled and stained with hematoxylin and eosin (HE) to investigate the histopathological changes of mouse lymph nodes in the infection group. The expression and localization of T lymphocyte surface markers CD3, CD4, and CD8 were observed in mouse lymph nodes using immunohistochemical staining. In addition, lymphocyte suspensions were prepared from mouse lymph nodes in both groups at different time points post-infection, and the levels of Tm cell subsets and their secreted cytokines were detected using flow cytometry. Results HE staining showed diffuse structural alterations in the subcapsular cortical and paracortical regions of mouse lymph nodes in the infection group 4 weeks post-infection with E. multilocularis. Immunohistochemical staining detected CD3, CD4 and CD8 expression in mouse lymph nodes in both groups. Flow cytometry revealed higher proportions of CD4⁺ Tm cells [(55.3 ± 4.8)% vs. (38.8 ± 6.1)%; t = -4.259, P < 0.05] and CD4⁺ tissue-resident Tm (Trm) cells [(57.7 ± 3.7)% vs. (34.1 ± 11.2)%; t = -3.990, P < 0.05] in mouse lymph nodes in the infection group than in the control group 4 weeks post-infection, and higher proportions of CD4⁺ Tm cells [(34.6 ± 3.2)% vs. (23.3 ± 7.5)%; t = -2.764, P < 0.05] and CD4⁺ Trm cells [(44.0 ± 1.9)% vs. (31.2 ± 1.5)%; t = -4.039, P < 0.05] in mouse lymph nodes in the infection group than in the control group 24 weeks post-infection. The proportions of CD8⁺ Tm cells were higher in the infection group than in the control group 4 weeks [(56.8 ± 2.7)% vs. (43.9 ± 5.2)%; t = -4.416, P < 0.01] and 12 weeks post-infection [(25.4 ± 2.7)% vs. (12.0 ± 2.6)%; t = -2.552, P < 0.05], while the proportions of tumor necrosis factor (TNF)-α⁺ CD4⁺ T cells [(15.7 ± 5.0)% vs. (49.4 ± 6.4)%; t = 7.150, P < 0.01], TNF-α⁺CD8⁺ T cells [(20.7 ± 5.5)% vs. (57.5 ± 8.4)%; t = -6.694, P < 0.01], and TNF-α⁺ CD8⁺ Tm cells [7.0% (1.0%) vs. 31.0% (11.0%); Z = -2.236, P < 0.05] were lower in the infection group than in the control group 24 weeks post-infection. Conclusions Tm cells levels are consistently increased in lymph nodes of mice at different stages of E. multilocularis infection, with Trm cells as the predominantly elevated subset. The impaired capacity of CD8⁺ Tm cells to secrete the effector molecule TNF-α in mouse lymph nodes at the late-stage infection may facilitate chronic parasitism of E. multilocularis.

Quercetin can mediate the activation of mitogen-activated protein kinase(MAPK)signaling pathways to prevent osteoporosis(OP).This paper comprehensively discusses the interrelationship between MAPK and osteoporosis-related cells based on the latest domestic and international research.Additionally,it elucidates the research progress of quercetin in mediating the MAPK signaling pathway for OP prevention.The aim is to provide an effective foundation for the clinical prevention and treatment of OP and the in-depth development of quercetin.

OBJECTIVE To evaluate the comprehensive quality of Houttuynia cordata from different producing areas. METHODS Using total flavonoids， water-soluble extract， moisture， total ash and acid-insoluble ash as indicators， the entropy weight method was used to objectively weigh each indicator， and the relative correlation degree （r）i calculated by grey correlation method was used as a measure to comprehensively evaluate the quality of H. cordata. RESULTS The weights of total flavonoids， total ash， moisture， acid-insoluble ash， and water-soluble extract were 0.295 5， 0.227 3， 0.188 7， 0.145 1， and 0.143 4， respectively. The weights of total flavonoids and total ash were relatively large. The ri of 30 batches of H. cordata ranged from 0.233 2 to 0.673 9； the average ri of samples from Quanzhou County and Ziyuan County of Guangxi Zhuang Autonomous Region were the highest， which were 0.638 3 and 0.598 7， respectively， followed by samples from Lingchuan County of Guangxi Zhuang Autonomous Region （0.556 1） and Jianshui County of Yunnan Province （0.452 8）. The quality of medicinal materials produced in the above producing areas was generally good and stable. CONCLUSIONS Entropy weight method combined with the grey correlation method can be used to comprehensively evaluate the quality of H. cordata. The overall quality of H. cordata produced in Quanzhou County of Guangxi Zhuang Autonomous Region is the best.

The n-butanol fraction of Alpinia oxyphylla Fructus 70% ethanol extract was separated and purified using column chromatography with MCI Gel CHP-20, Sephadex LH-20, ODS, and silica gel, combined with semi preparative liquid phase and TLC separation methods. One new halogenated 4,5-seco-eudesmane sesquiterpenoid and two new eremophilane sesquiterpenoids were isolated and purified from the n-butanol fraction of Alpinia oxyphylla Fructus. The structures of the isolated compounds were identified using modern spectroscopic methods (1D, 2D NMR, UV, IR, MS, etc.), and the absolute configuration of the new compounds were determined using the methods of calculated ECD and induced ECD.

Six compounds were isolated from the roots of Ephedra sinica Stapf using various chromatographic techniques such as silica gel column chromatography, thin layer chromatography and semi-preparative HPLC. Their chemical structures were identified by analysis of physicochemical properties and spectral data, and determined as (Z)-docosanylferulate (1), (E)-docosanylferulate (2), bis (2-ethylheptyl) phythalate (3), 2,2′-oxybis (1,4-di-tert-butylbenzene) (4), diisobutyl phthalate (5), bis (2-ethylhexyl) phthalate (6). Among them, compound 1 is a new compound, compounds 2-4 were first isolated from Ephedra. A corticosterone-induced PC-12 cell injury model was used for compound activity screening. The results showed that compounds 1 and 5 significantly improved corticosterone-induced PC-12 cell injury and significantly increased 5-HT₇ receptor protein expression in the cells, indicating potential antidepressant activity.

BACKGROUND:At present,there is still controversy in clinical practice about the choice of internal fixation of Pauwels type Ⅲ femoral neck fracture,and the selection of internal fixation that provides stable fixation strength is the key basis for achieving Pauwels type Ⅲ fracture fixation. OBJECTIVE:The three-dimensional finite element analysis method was used to test the difference in biomechanical strength of three types of internal fixation in Pauwels type Ⅲ femoral neck fracture,which provided a reference for its clinical treatment. METHODS:Using the CT data of the left femur of a healthy male volunteer,a complete femur and its cancellous bone were reconstructed in Mimics software,and Geomagic studio software was used for reverse modeling.Cannulated compression screw,dynamic hip screw,and femoral neck system were created in UG-NX software.Three kinds of internal fixation models were assembled on the femur model,and Pauwels type Ⅲ femoral neck fracture was simulated by Hypermesh software.Finally,Abaqus software was used to carry out finite element experimental analysis to analyze and compare the stress distribution,stress peak,strain,and displacement distribution caused by fixed femoral neck fracture of different internal fixation systems. RESULTS AND CONCLUSION:(1)The stress of the proximal femur bone mass was mainly distributed in the area below the femoral neck near the fracture end,with the highest stress peak in the dynamic hip screw group and the smallest in the femoral neck system group.(2)The stress distribution of the internal fixation device was mainly concentrated on the screw surface near the fracture line,with the highest stress peak in the femoral neck system group and the smallest in the dynamic hip screw group.(3)The main strain field of the proximal femur bone mass was distributed in the upper surface area where the bone and screw contacted,and the yield strain was the smallest in the femoral neck system group and the largest in the cannulated compression screw group.(4)The main strain field of the internal fixation device model was distributed on the upper surface of the femoral neck screw,with the yield strain being the smallest in the femoral neck system group and the largest in the cannulated compression screw group.(5)The displacement distribution values of femur,proximal bone mass,distal bone block,internal fixation device and internal fixation with the femur as a whole in the three femoral neck fracture internal fixation models decreased gradually from proximal to distal,and the peak displacement of the femoral neck system group was the largest and the lowest in the dynamic hip screw group.(6)The results showed that when the Pauwels type Ⅲ femoral neck fracture was fixed,the stress distribution of femoral neck system was more uniform,the mechanical conduction characteristics were better,and it was subjected to lower yield strain,higher stress and higher displacement.It has relatively better biomechanical stability and can provide a superior mechanical environment for fracture healing.

Objective To explore the effect of shikonin on the recovery of nerve function after acute spinal cord injury(SCI)in rats.Methods 96 male Sprague-Dawley(SD)rats were divided into 4 groups randomly:sham operation group(Group A),sham operation+shikonin group(Group B),SCI+DMSO(Group C),SCI+shikonin group(Group D).The acute SCI model of rats was made by clamp method in groups C and D.After subdural catheterization,no drug was given in group A.rats in groups B and D were injected with 100 mg·kg-1 of shikonin through catheter 30 min after modeling,and rats in group C were given with the same amount of DMSO,once a day until the time point of collection tissue.Basso-Beattie-Bresnahan(BBB)scores were performed on 8 rats in each group at 6,12,and 3 d after moneling,and oblique plate tests were performed on 1,3,7 and 14 d after modeling,and then spinal cord tissues were collected.Eight rats were intraperitoneally injected with propidine iodide(PI)1 h before sacrificed to detection PI positive cells at 24 h in each group.Eight rats were sacrificed in each group at 24 h after modeling,the spinal cord injury was observed by HE staining.The Nissl staining was used to observe survivor number of nerve cells.Western-blot technique was used to detect the expression levels of Bcl-2 protein and apoptosis related protein RIPK1.Results After modeling,BBB scores were normal in group A and B,but in group C and D were significantly higher than those in group A and B.And the scores in group D were higher than those in group C in each time point(P＜0.05).At 12 h after modeling,the PI red stained cells in group D were significantly reduced compared with that in group C,and the disintegration of neurons was alleviated(P＜0.05).HE and Nissl staining showed nerve cells with normal morphology in group A and B at 24h after operation.The degree of SCI and the number of neuronal survival in group D were better than those in group C,the differ-ence was statistically significant at 24h(P＜0.05).The expression of Bcl-2 and RIPK1 proteins was very low in group A and B;The expression of RIPK1 was significantly increased in Group C and decreased in Group D,with a statistically significant dif-ference(P＜0.05);The expression of Bcl-2 protein in group D was significantly higher than that in group C(P＜0.05).Conclu-sion Shikonin can alleviate the pathological changes after acute SCI in rats,improve the behavioral score,and promote the re-covery of spinal nerve function.The specific mechanism may be related to the inhibition of TNFR/RIPK1 signaling pathway mediated necrotic apoptosis.

Objective:To explore the application of viral load (VL) testing in human immunodeficiency virus (HIV) antibody screening positive but western blot (WB) negative samples, provide scientific basis for laboratory diagnosis of such populations.Methods:For the HIV antibody retest samples from the initial screening laboratory of Zaozhuang City Center for Disease Control and Prevention from 2021 to 2023, as well as samples from the voluntary counseling and testing at outpatient clinic of our unit that showed a negative WB test result, VL and CD4 + T lymphocyte counts were tested. Four weeks later, the patients were followed up and blood samples were collected by the current district (city) Center for Disease Control and Prevention, and sent to the confirmation laboratory for confirming testing. Results:Among the thirty-two patients, there were fifteen cases with target not detected (TND) by using VL. After four weeks, the follow-up results were all negative. There was one case with a VL test result of 20-5 000 copies/ml, and after four weeks, the follow-up result was positive. There were sixteen cases with a VL test result of >5 000 copies/ml. After four weeks, the follow-up results were all positive. Person correlation analysis was conducted between the log value of VL and CD4 + T lymphocyte count, and the result showed a certain degree of negative correlation ( r=-0.63, P=0.006). Conclusions:VL testing can assist in distinguishing patients with positive HIV antibody screening and negative WB results, and VL testing and WB testing should complement each other, combined with CD4 + T lymphocyte count and epidemiological history to make a diagnosis.

Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laboratory diagnosis of viral encephalitis is a worldwide challenge.Recently,high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections.Thus,In this study,we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing. Methods We designed nine pairs of specific polymerase chain reaction(PCR)primers for the 12 viruses by reviewing the relevant literature.The detection ability of the primers was verified by software simulation and the detection of known positive samples.Amplicon sequencing was used to validate the samples,and consistency was compared with Sanger sequencing. Results The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×,and the sequence lengths were consistent with the sizes of the predicted amplicons.The sequences were verified using the National Center for Biotechnology Information BLAST,and all results were consistent with the results of Sanger sequencing. Conclusion Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis.It is also a useful tool for the high-volume screening of clinical samples.

Objective To analyze the changes of B lymphocyte(B cells)subsets in peripheral blood of patients with chronic hepatitis B(CHB)and to explore its clinical significance.Methods Peripheral blood samples were collected from 37 treatment-na?ve CHB patients who were admitted to the Fifth Medical Center of PLA General Hospital from July 2022 to October 2022,and peripheral blood samples collected from 18 healthy individuals who have received the hepatitis B vaccine as healthy controls(HC).The study subjects'clinical indexes such as age,HBV DNA viral load,HBsAg quantification,HBeAg semi quantification,ALT,AST,and AST/ALT ratio were collected.The change characteristics of the frequency,phenotypic and functional markers of peripheral blood B lymphocytes and their subsets were compared between CHB and HC.Using multi-color flow cytometry,and the correlation between them and clinical indexes was analyzed.Results Frequency analysis of each subset of B cells showed that compared with HC,the frequency of total B cells,transitional B cells and naive B cells was decreased(P<0.05),while the frequency of mature B cells,memory B cells,atypical memory B cells and activated memory B cells was increased in CHB patients(P<0.01).And there was no significant difference in the frequency of resting memory B cells between the two groups(P>0.05).The results of functional analysis showed that compared with HC,the expression levels of CD79b on total B cells,mature B cells,memory B cells,naive B cells,activated memory B cells,atypical memory B cells and resting memory B cells in CHB patients were increased(P<0.05).The expression level of programmed cell death protein-1(PD-1)on atypical memory B cells in CHB patients was also higher than that in HC group(P<0.05).The results of correlation analysis showed that the frequency of total B cells in CHB patients was slightly negatively correlated with age(r=-0.39,P<0.05),while the expression of programmed death-1(PD-1)on total B cells,mature B cells,transitional B cells,memory B cells and naive B cells were slightly positively correlated with age(r>0.36,P<0.05).Conclusions Chronic HBV infection leads to depletion of the frequency and function of a portion of B cells in the peripheral blood of CHB patients,and age is a potential risk factor for the decline in humoral immune function in CHB patients.