ObjectiveTo establish a quality evaluation method for Zhuye Shigao granules（Zhuye Shigaotang） based on fingerprint and determination of index components， and to investigate the therapeutic effect of Zhuye Shigao granules on mice with cold-dampness pestilence attacking lung syndrome. MethodsThe fingerprint of Zhuye Shigao granules was established by high performance liquid chromatography（HPLC）， and the methods for determination of total calcium， orientin， isoorientin， ginsenosides Rg₁， Re and Rb₁ and other 2 index components were established. Fifty ICR mice were randomly divided into the blank group， model group， Zhuye Shigao granules low， medium and high dose groups（9.3， 18.6， 37.2 g·kg^-1·d^-1）， with 10 mice in each group. In addition to the blank group， the model mice with cold-dampness pestilence attacking lung syndrome was prepared by nasal drip of lipopolysaccharide combined with cold-dampness environment. Each administration group was given the corresponding liquid by gavage according to the dose， while the blank group and model group were given the same volume of normal saline by gavage. Then， the body temperature and organ index of mice in each group were measured， hematoxylin-eosin（HE） staining was used to investigate the lung tissue injury of mice in each group， and enzyme-linked immunosorbent assay（ELISA） was used to detect the changes of tumor necrosis factor-α（TNF-α）， interleukin（IL）-lβ， IL-6， IL-10 levels in serum and lung tissue， as well as immunoglobulin（Ig）A and IgM levels in serum. ResultsThe fingerprint similarity of 10 batches of Zhuye Shigao granules was>0.950， and 20 common peaks were calibrated. Seven of them were identified， including peak 11（isoorientin）， peak 12（orientin）， peak 14（apioside liquiritin）， peak 15（liquiritin）， peak 17（apioside isoliquiritin）， peak 19（isoliquiritin） and peak 20（liquiritigenin）. The results of quantitative analysis showed that the content range of each index component in 10 batches of Zhuye Shigao granules was as follows：Total calcium of 9.978-11.294 mg·g^-1， isoorientin of 0.033-0.041 mg·g^-1， orientin of 0.046-0.055 mg·g^-1， ginsenoside Rg₁+ginsenoside Re of 0.748-0.762 mg·g^-1， ginsenoside Rb₁ of 0.151-0.197 mg·g^-1， liquiritin of 1.106-1.366 mg·g^-1， glycyrrhizic acid of 0.904-1.182 mg·g^-1_. Compared with the blank group， the body temperature of mice in the model group was significantly increased， the organ indexes of liver， lung and spleen were significantly decreased， the organ index of thymus was significantly increased， HE staining of lung tissue showed infiltration of inflammatory cells， a small amount of serous exudation was observed in the alveoli， and lung tissue was damaged. After the intervention of Zhuye Shigao granules， the pathological changes were improved compared with the model group. The expression levels of IL-1β， IL-6 and TNF-α were significantly increased， the expression level of IL-10 was significantly decreased in serum and lung tissue. The levels of IgA and IgM in serum were significantly decreased（P<0.01）. Compared with the model group， the body temperature， the organ indexes and immune factor levels in serum and lung tissue of mice in the Zhuye Shigao granules medium and high dose groups were significantly reduced（P<0.05， P<0.01）. ConclusionIn this study， the quality evaluation of Zhuye Shigao granules was carried out based on fingerprint combined with determination of index components， and the fingerprint of four herbs（Lophatheri Herba， Ophiopogonis Radix， Pinelliae Rhizoma and Glycyrrhizae Radix et Rhizoma） in this formula and the determination of 8 index components were established. The therapeutic effect of Zhuye Shigao granules on mice with cold-dampness pestilence attacking lung syndrome may be related to inhibiting inflammatory response and mediating immune regulation.

Objective:To observe the improving effects of Tuina(Chinese therapeutic massage)plus exercise therapy on joint alignment and walking function in children with genu varum(GV).Methods:Sixty-six children with GV were divided into an exercise therapy group and a Tuina plus exercise therapy group using the random number table method,with 33 cases in each group.Both groups received identical exercise therapy,while the Tuina plus exercise therapy group was additionally offered Tuina manipulation treatment.The intervention course lasted 12 weeks in both groups.Before and after the intervention,the three-dimensional gait analysis was adopted to assess the spatiotemporal parameters,kinematics,and kinetic characteristics of lower-limb joints in children with GV.Results:The GV angle was reduced after intervention in the Tuina plus exercise therapy group(P<0.05),but there was no significant change in the exercise therapy group(P>0.05).After treatment,the Tuina plus exercise therapy group demonstrated a notable decrease in the step length,walking speed,peak forefoot adduction angle,and peak ankle inversion moment(P<0.05),suggesting the correction of the ankle joint's compensatory pathological changes.In the exercise therapy group,the foot progression angle(FPA)and gait deviation index(GDI)increased markedly after the intervention(P<0.05),indicating improved overall kinematic function.Conclusion:The combined use of Tuina manipulations and exercise therapy can produce significant effects in correcting the knee joint's torsion,both coronally(GV angle)and horizontally(ankle inversion moment,FPA,and forefoot adduction angle),while exercise therapy alone can markedly improve the overall kinematic parameters(FPA and GDI).

Objective To investigate the distribution of traditional Chinese medicine(TCM)syndrome types in patients with rheumatoid arthritis(RA)and to explore their correlation with serum homocysteine(Hcy)and C-reactive protein(CRP)levels.Methods From December 2021 to December 2023,a total of 186 RA patients admitted to Dezhou Hospital of Qilu Hospital of Shandong University were selected as the study group,and the patients were classified into four TCM syndrome types according to the criteria of TCM syndrome differentiation,namely cold-damp blockage syndrome,qi and blood deficiency syndrome,phlegm-stasis blockage syndrome,deficiency of liver and kidney syndrome.Moreover,100 healthy volunteers who had physical examination at the same hospital during the same period were included as the healthy control group.The distribution of TCM syndrome types in RA patients was investigated,the serum Hcy and CRP levels in the two groups and in RA patients with various TCM syndrome types were observed,and then Pearson's correlation was used for explorating the correlation of serum Hcy and CRP levels with severity of illness of RA patients with different TCM syndrome types.Results(1)Among the 186 RA patients in the study group,there were 41 cases(22.04％)of cold-damp blockage syndrome,40 cases(21.51％)of phlegm-stasis blockage syndrome,50 cases(26.88％)of qi and blood deficiency syndrome,and 55 cases(29.57％)of deficiency of liver and kidney syndrome.(2)The serum Hcy and CRP levels in the study group were(7.61±2.07)μmol/L and(27.93±5.92)mg/L,respectively,which were higher than those of the healthy control group[(5.33±0.91)μmol/L and(1.85±0.67)mg/L,respectively],and the differences were statistically significant(P<0.01).Statistically significant differences of serum Hcy and CRP levels were shown among the patients with various TCM syndrome types(F=7.637,P<0.01;F=5.639,P<0.01).The serum levels of Hcy and CRP in patients with cold-damp blockage syndrome were the highest,and the levels in patients with phlegm-stasis blockage syndrome,qi and blood deficiency syndrome,and deficiency of liver and kidney syndrome were in decreasing sequence.The pairwise comparison between groups showed that the differences were statistically significant(P<0.05).(3)Pearson's correlation analysis showed that Disease Activity Score in 28 joints(DAS28)scores were positively correlated with serum Hcy and CRP levels(r=0.396,P<0.01;r=0.407,P<0.01),and the serum Hcy and CRP levels were positively correlated with the severity of illness in RA patients with different TCM syndrome types(P<0.05 or P<0.01).Conclusion The expression of serum Hcy and CRP in patients with RA is abnormally increased,and the expression levels of serum Hcy and CRP are correlated with TCM syndrome types.RA patients with cold-damp blockage syndrome have the highest expression levels of serum Hcy and CRP.

Objective To compare the clinical value of TruScreen,an artificial intelligence-based real-time cervical cancer screening system,combined with high-risk human papillomavirus(hr-HPV)test and Thinprep cytology test(TCT)combined with hr-HPV test in cervical cancer screening.Methods A total of 297 women undergoing cervical cancer screening at Panzhihua Central Hospital between June 2020 and December 2023 were enrolled.All participants underwent hr-HPV testing,TCT,and TruScreen test.The diagnostic performance of TruScreen combined with hr-HPV test and TCT combined with hr-HPV test for low-grade squamous intraepithelial lesion positive(LSIL+)and high-grade squamous intraepithelial lesion positive(HSIL+)cervical lesions was evaluated.Results Among the 297 enrolled women,pathology analysis identified 128(43.10%)LSI+cases and 67(22.56%)HSIL+cases.Additionally,there were 110(37.04%)HPV16/18 positive cases,177(59.60%)cases with TCT results≥atypical squamous cells of undetermined significance(ASCUS),and 176(59.26%)cases with abnormal TruScreen results.The area under the curve(AUC)of TruScreen combined with hr-HPV test in diagnosing LSIL+and HSIL+cervical lesions was higher than that of TCT combined with hr-HPV test(P<0.05).Conclusion TruScreen combined with hr-HPV test demonstrates superior performance in cervical cancer screening compared with TCT combined with hr-HPV test.TruScreen combined with hr-HPV test may serve as an alternative to conventional cytology-based methods for cervical cancer screening in China.

Objective:To investigate the status quo of the salary status of Guangxi's village doctors under the background of the implementation of"Employed by Townships and Work in the Villages"Policy.Methods:A total of 142 village doctors'salary data were obtained and sorted out through field investigation.Village doctors'salary level was analyzed with Chi-square test,rank test and correlation analysis.Results:More than 70%of village doctors'annual salary in the surveyed counties exceeded 50 000 yuan.Performance allowance is the main component of village doctors'annual salary.There are differences between the implementation number of"Employed by Townships and Work in the Villages"Policy and village doctors'annual salary level in different urban areas(P＜0.05).The village doctors'service population was positively correlated with the allowance of basic public health service,the general medical service population and the annual salary level(P＜0.05).Conclusion:Under the"Employed by Townships and Work in the Villages"Policy,the salary level of Guangxi's village doctors has been improved,and the salary component could be further optimized.It is suggest that the"Employed by Townships and Work in the Villages"Policy should be promoted in accordance with the economic development conditions of different urban areas,the income standards that match village doctors'academic,professional ranks,professional qualifications and working years,and the performance evaluation standards linked to village doctors'subsidy standards should be improved.

ObjectiveTo clarify the neuroprotective effect of black Panacis Quinquefolii Radix（PQR） and explore its active ingredients， with the aim of establishing an activity-oriented quality evaluation method. MethodsTransgenic Tg（HuC∶EGFP） zebrafish was used to establish a neuronal injury model by aluminum chloride immersion. Different doses（10， 20 mg·L^-1） of PQR and black PQR ethanol extracts were administered. The neuroprotective effects of PQR and black PQR were compared by analyzing the fluorescent area and intensity of zebrafish neurons. Based on ultra-performance liquid chromatography（UPLC）， a fingerprint profile of black PQR was established， followed by principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA）. Differential components were screened using the criteria of variable importance in the projection（VIP） value>1 and P<0.05. The neuroprotective activity of 14 batches of black PQR was assessed， and Spearman correlation analysis was used to identify saponins related to neuroprotective activity， which were then validated. Based on the above results， active marker components were determined， and an UPLC method was established for their quantitation with clear content limits. ResultsPharmacological efficacy results showed that both PQR and black PQR at different doses could significantly improved neuronal damage in zebrafish. At a dose of 20 mg·L^-1， black PQR demonstrated superior efficacy（P<0.05）. The fingerprint similarities of 14 batches of black PQR were>0.94， with 26 common peaks identified. Through comparison with the reference standards， 8 components were confirmed， including peak 1（ginsenoside Rg₁）， peak 2（ginsenoside Re）， peak 5（ginsenoside Rb₁）， peak 9（ginsenoside Rd）， peak 16［ginsenoside 20（S）-Rg₃］， peak 17［ginsenoside 20（R）-Rg₃］， peak 18（ginsenoside Rk₁）， and peak 19（ginsenoside Rg₅）. The results of PCA and OPLS-DA indicated that there were differences in saponins among black PQR samples from different origins， and 12 differential components were screened. All 14 batches of black PQR exhibited good protective effects on zebrafish neurons， with Shaanxi-produced black PQR showing superior protective effects compared to the other three production regions. Spearman correlation analysis revealed that a total of 11 components， including ginsenosides 20（S）-Rg₃， 20（R）-Rg₃， Rk₁ and Rg₅， showed a significant positive correlation with the neuroprotective effect in zebrafish（P<0.05）. The activity validation results indicated that ginsenosides 20（S）-Rg₃， 20（R）-Rg₃， Rk₁ and Rg₅ were the primary components responsible for the neuroprotective effects of black PQR. Quantitative analysis showed that the content of ginsenoside 20（S）-Rg₃ in 14 batches of black PQR ranged from 0.17% to 0.52%， and the repair rate of neuronal damage ranged from 42.77% to 97.83%. ConclusionBased on the fingerprint and neuronal protective activity， the spectrum-effect related quality control model of black PQR was established， with ginsenoside 20（S）-Rg₃ as the quality control index， and the neuronal damage repair rate≥60% as the evaluation standard， the minimum limit of ginsenoside 20（S）-Rg₃ in black PQR should be≥0.20%.

Objective: To investigate the predictive value of EZH2 expression for malignant transformation in oral leukoplakia (OLK) and to provide a reference for clinical practice. Methods: This study was approved by the institutional ethics committee, and informed consent was obtained from all participants. A total of 114 patients diagnosed with OLK by pathological examination and treated at our hospital between November 2020 and July 2022 were initially enrolled. After excluding those with incomplete data or follow-up, 105 participants were included in the final analysis, comprising 14 in the high EZH2 expression group and 91 in the low EZH2 expression group. Histopathological examination of oral mucosa and immunohistochemical detection of EZH2 protein expression were performed. The follow-up period was 30 months; participants were followed until malignant transformation occurred or until the end of follow-up, at which point they were withdrawn from the study. The exposure factor was the level of EZH2 protein expression, and the outcome was the malignant transformation rate of OLK. Differences in EZH2 expression levels and transformation outcomes were analyzed. Results: There were no statistically significant differences between the high and low EZH2 expression groups in terms of age, sex, history of systemic disease, lifestyle habits, psychological status, diet, and sleep conditions (P > 0.05). Lesions in the high EZH2 expression group were mainly located on the ventral tongue, while in the low EZH2 expression group, they were more commonly found on the dorsal tongue and buccal mucosa. The malignant transformation rate was 28.6% (4/14) in the high expression group and 8.8% (8/91) in the low expression group; these differences were not statistically significant (P=0.053). In univariate Cox regression analysis, the risk of malignant transformation in the high EZH2 expression group was 3.647 times that of the low EZH2 expression group (HR = 3.647, 95% CI: 1.097-12.120, P<0.05). Kaplan-Meier survival analysis showed that over the 30-month follow-up period, the cancer-free survival rate in the high EZH2 expression group was 19.8% lower than in the low expression group, and the difference was statistically significant (P<0.05). In multivariate Cox regression analysis, only moderate and severe epithelial dysplasia were identified as independent risk factors for malignant transformation. The risk of malignant transformation in the moderate and severe dysplasia groups was 10.695 and 13.623 times higher, respectively, than in the mild dysplasia group (HR = 10.695, 95% CI: 2.270-50.396, P<0.05; HR=13.623, 95% CI: 1.918-96.774, P<0.05). EZH2 high expression was not an independent risk factor in the multivariate model (HR= 2.528, 95% CI: 0.752-8.500, P = 0.134). Conclusion High EZH2 protein expression is a risk factor for the malignant transformation of OLK but does not have independent predictive value.

Objective:To apply mass spectrometry as well as bioinformatics techniques to analyze HBV peptides derived from Pol and X proteins presented by human immortalized B lymphocytes(BLCLs),and to screen for effective T-cell epitopes,which lays the foundation for the development of therapeutic vaccines.Methods:The group has constructed a genome-wide expression plasmid containing 1.2-fold HBVC2 isoforms and transfected it into BLCLs by electro-transfection,isolated and identified HBV peptides by LC-MS/MS,bioinformatically predicted peptide sequences in terms of sensitization,antigenicity,toxicity,HLA molecular affinity,and the ability of peptide HLA-class Ⅰ complex to bind to specific T cells,and retrieved reported sequences.Results:HBV peptides from lysates of five immortalized B cells(BLCLs-1 to BLCLs-5)carrying HBVC2 subtype-expressing recombinants were analyzed,and 141 peptides with sequence lengths of no less than 8 amino acid residues(≥8 aa)were successfully isolated and identified,of which 133 were derived from Pol,and 8 from X proteins.The 141 HBV peptides were analyzed for sensitization,antigenicity and toxicity,and 50 antigenic,non-toxic and sensitizing HBV peptides(47 from Pol and 3 from X protein)were screened for affinity analysis with HLA-class Ⅰ and Ⅱ molecules and for prediction of the binding ability of the peptide HLA-class Ⅰ complexes to specific T cells.Among these peptides,37 had affinity to the corresponding genotypes of HLA-class Ⅰ and Ⅱ molecules.Finally,we obtained 37 peptides with antigenic,nontoxic,and sensitizing properties with IC50<500 nmol/L to HLA-class Ⅰ and Ⅱ molecules,which have affinitied to the corresponding genotypes but have not been reported,and can be continued to be explored as potential epitopes.Finally,15 HBV peptide hotspot core regions were formed by intra-and inter-strain common,contained,overlapping or neighboring sequence analysis of 141 peptides,with 7 core region sequences with antigenic,nontoxic,and sensitizing properties restricted to the corresponding geno-types.Peptides with affinity IC50<500 nmol/L had an affinity core sequence overlap of no less than 8 amino acids,and can be priori-tized for candidate T cell epitopes for subsequent studies.Conclusion:Peptides derived from Pol and X proteins have been successfully isolated and identified,and potential T cell epitopes have been screened.

Objective To construct a risk prediction model for atherosclerosis(AS)in patients with rheumatoid arthritis(RA)based on Lasso-Logistic regression analysis and provide a scientific basis for individualized clinical intervention.Methods The retrospective clinical data were collected from 344 RA patients,including 86 patients with AS(RA+AS group)and 258 patients with without AS(RA group).The clinical characteristics and initial laboratory test results were compared between the two groups.Lasso regression was used to screen the key predictive variables,and Logistic regression was combined to construct the prediction mode.The discrimination of the model was evaluated through the receiver operating characteristic(ROC)curve and the area under the curve(AUC).The Hosmer-Lemeshow test was used to assess the calibration,and decision curve analysis was used to verify the clinical applicability of the model.Results Seven predictive variables were identified including RA disease duration,DAS28 score,C-reactive protein(CRP),triglycerides(TG),high-density lipoprotein cholesterol(HDL-C),fasting blood glucose(FBG)and hypertension.The risk prediction model for AS in RA patients was:Logit(P)=-2.674+0.605×RA disease duration+0.393×DAS28 score+0.310×CRP+1.346×TG-2.289×HDL-C+0.679×FBG+0.711×hypertension.The AUC of the model was 0.965(95%CI:0.943-0.987),and the Hosmer-Lemeshow test showed χ2=0.547,P=1.000,indicating good discrimination and calibration.Clinical decision curve analysis showed that the probability threshold ranged from 7%to 92%,demonstrating high clinical applicability.Conclusion The AS risk prediction model constructed in this study for RA patients can effectively identify high-risk individuals,supporting the development of personalized prevention and treatment strategies.

Objective To investigate the gait characteristics of school-age children with generalised joint hypermobility(GJH).Methods The kinematic and kinetic data of lower extremity joints in 56 children with GJH and 56 healthy children were collected by 3D motion capture system and 3D force plates,and the differences between the two groups were compared and analyzed.Results Kinematics parameters:the mean ankle inversion angle(P=0.000),maximum hip adduction angle(P=0.002)and maximum foot clearance angle(P=0.004)in stance phase decreased in the GJH children group,while the mean foot progression angle(P=0.000),the maximum foot internal rotation angle(P=0.000)in stance phase,and the knee flexion angle(P=0.032)in the middle stance phase increased.Kinetics parameters:the maximum power of knee(P=0.005)and hip(P=0.000)joints and the maximum inversion moment of ankle joints(P=0.009)in stance phase decreased in GJH children group.Temporal and spatial parameters:the step length(P=0.001),walking velocity(P=0.000),and cadence(P=0.000)decreased in GJH children group,while the stance phase percentage(P=0.000),and step width(P=0.000)increased.Conclusions There is a decrease in walking efficiency and balance stability of children with GJH.The line alignment is abnormal in ankle joint.The knee function needs a long-term attention.