Osteoarthritis （OA） and stroke are common clinical diseases that reduce patients' quality of life and place a burden on families and society. Ruyi Zhenbaowan， a classic prescription in Tibetan medicine， have the functions of clearing heat， awakening the brain and opening orifices， relaxing tendons and promoting meridian circulation， and eliminating yellow water. Clinically， they are used to treat osteoarthritis， post-stroke sequelae， neuropathic pain， and other related conditions. Modern pharmacological studies have demonstrated their anti-inflammatory， analgesic， and nerve-repairing effects. However， current research remains insufficient regarding the appropriate indications， timing， and efficacy of this medicine in treating relevant diseases. To enhance clinicians' understanding of this medicine and promote its standardized and rational clinical use， a panel of national experts， including clinical specialists， Tibetan medicine practitioners， pharmacologists， and methodologists， formulated this consensus based on clinical experience and evidence-based practice. The Cochrane systematic review framework， the Grading of Recommendations Assessment， Development and Evaluation （GRADE） system， and the nominal group method were employed to generate seven graded recommendations and 19 consensus-based suggestions. These recommendations clearly define the key points in the clinical application of Ruyi Zhenbaowan， including therapeutic indications， dosage and administration， treatment duration， and medication safety. The consensus specifically addresses the clinical efficacy， appropriate timing of administration， dosage strategies， treatment cycles， and combination medication strategies for treating osteoarthritis and stroke and provides an overview of safety considerations. The aim is to provide standardized guidance for hospitals and healthcare institutions nationwide to ensure the rational application of Ruyi Zhenbaowan in the treatment of osteoarthritis and stroke， reduce medication-related risks， and further leverage its clinical advantages. This consensus has been approved and issued by the China Association of Chinese Medicine， with the standard number GS/CACM 369-2024.

Objective To develop a reagent for detecting of 17α-hydroxyprogesterone(17α-OHP)in dried blood spots on filter paper fixed on vascular stent by Auto TRFIA-4 automatic fluorescence immunoanalyzer and evaluate its performance.Methods The microwell plate was coated with the sheep anti-rabbit IgG antibody as microwell reaction plate,the rabbit anti-human 17α-OHP antibody was diluted as intermediate antibody,and the 17α-OHP-bovine serum albumin conjugate was labeled by europium as europium marker.The concentration of 17α-OHP in dried blood spots on filter paper fixed on blood spots stent was quantitatively detected by Auto TRFIA-4 automatic fluorescence immunoanalyzer.The analysis sensitivity,accuracy,linearity,precision,specificity and stability were evaluated,and whether they met the requirements of the formulated industry standards were evaluated.A total of 227 neonatal heel blood filter paper samples from newborns who were born 72 hours after birth and within 7 days and fully breastfeeding were selected for reagent comparison test.The consistency was analyzed by χ2 test,Kappa test,t test,linear correlation analysis,regression analysis,Bland-Altman method analysis and predictive bias analysis of medical decision level,P<0.05 indicated statistically significant difference.Results The optimal coating concentration of sheep anti-rabbit IgG antibody was 3 μg/ml.The optimal dilution ratio of rabbit anti-human 17α-OHP antibody was 1∶1 500.The optimal dilution ratio of 17α-OHP europium marker mother liquor was 1∶2 500.The limit of blank,limit of detection,limit of quantification was 0.75,1.08 and 1.99 nmol/L,respectively.The relative deviations of the standard check test were within±15.00%,and the average recovery rate was 92.36%.The linear correlation coefficient was 0.997 1 in the range of 2.00 to 300.00nmo/L.The intra-assay and inter-assay coefficients of variation were all within 10.00%.The cross-reactivity rates of 100.00 ng/ml progesterone,17α-hydroxypreg nenolone and 11-deoxycortisol were within 0.089%to 0.64%.The performance of stability test met the requirements.The total coincidence rate was 100%compared with the results of contrast reagent.The quantitative results were highly correlated with the contrast reagent(r=0.999 4,tr=452.02,P<0.05).Conclusion The self-developed reagent has the advantages of high sensitivity,good accuracy,wide linear range,good precision,high specificity and good stability,which meets the requirements of the formulated industry standards,and has high correlation and consistency with the result of contrast reagent,which meets the needs of clinical detection.

Objective To develop a reagent for detecting of 17α-hydroxyprogesterone(17α-OHP)in dried blood spots on filter paper fixed on vascular stent by Auto TRFIA-4 automatic fluorescence immunoanalyzer and evaluate its performance.Methods The microwell plate was coated with the sheep anti-rabbit IgG antibody as microwell reaction plate,the rabbit anti-human 17α-OHP antibody was diluted as intermediate antibody,and the 17α-OHP-bovine serum albumin conjugate was labeled by europium as europium marker.The concentration of 17α-OHP in dried blood spots on filter paper fixed on blood spots stent was quantitatively detected by Auto TRFIA-4 automatic fluorescence immunoanalyzer.The analysis sensitivity,accuracy,linearity,precision,specificity and stability were evaluated,and whether they met the requirements of the formulated industry standards were evaluated.A total of 227 neonatal heel blood filter paper samples from newborns who were born 72 hours after birth and within 7 days and fully breastfeeding were selected for reagent comparison test.The consistency was analyzed by χ2 test,Kappa test,t test,linear correlation analysis,regression analysis,Bland-Altman method analysis and predictive bias analysis of medical decision level,P<0.05 indicated statistically significant difference.Results The optimal coating concentration of sheep anti-rabbit IgG antibody was 3 μg/ml.The optimal dilution ratio of rabbit anti-human 17α-OHP antibody was 1∶1 500.The optimal dilution ratio of 17α-OHP europium marker mother liquor was 1∶2 500.The limit of blank,limit of detection,limit of quantification was 0.75,1.08 and 1.99 nmol/L,respectively.The relative deviations of the standard check test were within±15.00%,and the average recovery rate was 92.36%.The linear correlation coefficient was 0.997 1 in the range of 2.00 to 300.00nmo/L.The intra-assay and inter-assay coefficients of variation were all within 10.00%.The cross-reactivity rates of 100.00 ng/ml progesterone,17α-hydroxypreg nenolone and 11-deoxycortisol were within 0.089%to 0.64%.The performance of stability test met the requirements.The total coincidence rate was 100%compared with the results of contrast reagent.The quantitative results were highly correlated with the contrast reagent(r=0.999 4,tr=452.02,P<0.05).Conclusion The self-developed reagent has the advantages of high sensitivity,good accuracy,wide linear range,good precision,high specificity and good stability,which meets the requirements of the formulated industry standards,and has high correlation and consistency with the result of contrast reagent,which meets the needs of clinical detection.

Autologous ozonized blood transfusion(AOBT) is a therapy of re-transfusion of 100-200 mL of autologous blood after shaking and agitation with appropriate amount of oxygen-ozone in vitro. The oxidation of blood through the strong oxidation of ozone can enhance the non-specific immune response of the body, regulate the internal environment and promote health. This therapy has been increasingly applied in clinical practice, while no unified standard for the operation process in terms of ozone concentration, treatment frequency and treatment course had been established. This operation process of AOBT is primarily explored in order to standardize the operation process and ensure its safety and efficacy.

Objective Few clinical studies have been reported on the reversibility of uremic cardiomyopathy (UC) after renal transplantation. This article aimed to investigate the cardiac structure and function of end-stage renal disease (ESRD) patients undergoing renal transplantation using cardiac magnetic resonance (CMR). Methods This study included 38 ESRD patients undergoing renal transplantation in the National Clinical Research Center for Kidney Diseases, General Hospital of Eastern Theater Command, from September 2015 to February 2017. All the patients received initial CMR examination at 1－2 days before renal transplantation and during the postoperative follow-up. At the median follow-up time of 3.5 (3.4-3.7), 7.0 (3.7-9.5) and 8.4 (7.1-12.7) months, we recorded the CMR parameters, including the left ventricular end-diastolic volume (LVEDV), end-systolic volume (LVESV), end-diastolic mass (LVEDM), end-systolic mass (LVESM), ejection fraction (LVEF), and native myocardial T1 relaxation time, and compared the parameters obtained before and after surgery. Results Twenty-five of the patients completed the postoperative follow-up, who averaged 27.5 years of age, with no history of diabetes mellitus or ischemic heart disease, and treated by dialysis for 1.7 (1.5-2.2) years. At 7.0 months after renal transplantation, as compared with the baseline, the patients showed significant decreases in the LVEDV ([96.7 ± 22.8] vs [83.4 ± 17.4] mL/m², P < 0.05), LVESV ([44.3 ± 14.8] vs [33.0 ± 10.9] mL/m², P < 0.05) and LVEDM ([67.1 ± 24.2] vs [59.0 ± 17.0] mL/m², P < 0.05), but an increase in the LVEF ([54.1 ± 10.6] % vs [60.9 ± 9.6] %, P < 0.01). The LVEDV and LVESV were also remarkably lower at 3.5 and 8.4 months than the baseline (P < 0.001), and so were the left ventricular at basal, mid, apical and global native T1 relaxation times at 3.5, 7.0 and 8.4 months (P < 0.05). Conclusion For young ESRD patients with no history of diabetes mellitus or ischemic heart disease and on short-term dialysis, left ventricular dilatation, systolic dysfunction and diffuse myocardial fibrosis are reversible after renal transplantation. Native T1 relaxation time can be used as a sensitive indicator to evaluate the degree of diffuse myocardial fibrosis in ESRD patients.

Background:There is no specific therapy for inflammatory bowel disease(IBD),and the pathogenesis of IBD is not fully clear. Aims:To explore the expression and clinical significance of IL-17BR in colon mucosa and peripheral blood mononuclear cell(PBMC)of patients with IBD. Methods:Colon mucosal biopsy specimens of 40 Crohn's disease(CD) patients,32 ulcerative colitis(UC)patients and 25 healthy controls and PBMC of 30 CD patients,27 UC patients and 25 healthy controls were collected. The expressions of IL-17BR in colon mucosa and PBMC were determined by immunohistochemistry and flow cytometry,respectively,and correlations between IL-17BR expression and levels of CRP, ESR,CDAI score and Mayo score were analyzed. Serum levels of TNF-α before and after infliximab(IFX)treatment were determined by ELISA,and correlations with IL-17BR were analyzed. Results:Compared with healthy controls,IL-17BR expressions in colon mucosa of CD and UC patients were significantly increased(P<0.05). IL-17BR expressions were significantly higher in active CD and UC patients than in remission CD and UC patients(P <0.05). No significant differences in IL-17BR expression in PBMC were found among CD,UC patients and healthy controls(P>0.05). The expression of IL-17BR in colon mucosa was positively correlated with CRP,ESR,CDAI score or Mayo score in CD,UC patients(P <0.05). After treatment with IFX,expression of IL-17BR in colon mucosa and serum TNF-α level were significantly decreased in CD patients(P<0.01). Expression of IL-17BR was positively correlated with serum TNF-α level in CD patients(P<0.05). Conclusions:The increasing of IL-17BR expression in IBD patients is closely correlated with activity of inflammation and TNF-α level. IL-17BR may play a vital role in immune response of intestinal mucosa,and can be used as a new marker for reflecting the activity of IBD.

OBJECTIVETo evaluate the adjunctive effect of diode laser irradiation for the treatment of periodontal-endodontic combined lesions.

METHODS30 cases with periodontal-endodontic combined lesions were selected. All cases were randomly divided into experimental group and control group (n=15). In the control group, patients were treated with root canal therapy and conservative periodontal treatment (scaling and root planning), and patients in the experimental group were treated with root canal therapy and conservative periodontal treatment followed by diode laser irradiation. Clinical measurements including pocket probing depth (PPD), clinical attachment level (CAL), modified bleeding index (mBI), periapical index (PAI) were taken before treatment (baseline). Three and six months after the treatment, the adjunctive effects of diode laser in the treatment of periodontal-endodontic combined lesions were evaluated.

RESULTSThree months after the treatment, the average PPD, CAL and mBI levels were reduced significantly in both groups. But the average PPD reduction in the experimental group was significantly greater (P<0.05) than in control group. However the average CAL and mBI reduction were not significantly different between two groups. Six months after the treatment, the average PPD, CAL and mBI levels were further reduced in the experiment group, but not in the control group. The average PAI levels were reduced significantly in both groups, but the PAI reduction was not significantly different between two groups.

CONCLUSIONThe use of diode laser as an adjunct procedure in the treatment of periodontal-endodontic combined lesions can aid in achieving success.

To evaluate the relationship between p27Kip1 low expression in breast cancer and its prognostic implication in breast carcinoma patients. Methods All data that were associated with the study of the relationship between p27Kip1 and the prognosis for breast cancer was pooled from Cochrane library, PubMed, Embase and Medlinebase. The outcome was measured using the risk ratio (RR). Data pooling was performed by RevMan 4. 2. Results 6457 patients from 20 studies were included in this meta-analysis. RR estimate of overall survival (OS) for patients with low level p27Kip1 was 2.07 [1.66,2.60] (P<0.01). For disease free survival (DFS), the pooled RR was 1.27 [1.10,1.47] (P<0.05). The combined RR estimate of relapse free survival (RFS) for patients with low level of p27Kip1 was 1.49 [0.92, 2.42] (P >0.05). In patients with lymph node negative breast carcinoma, the combined RR for OS and RFS were 1.98 [1.34,2.91] (P <0.01) and 1.28 [0.45,3.65] (P > 0.05), respectively. Among the patients with lymph node positive breast carcinoma, the combined RR for OS and RFS was 1.92 [1.31, 2.82] (P=0.0009) and 1.35 [0.96,1.89] (P>0.05) respectively. Conclusions Low level of p27Kip1 appears to be an independent prognostic factor to OS and DFS of breast cancer patients but not to RFS. Additional studies with large patient number and widely accepted practical methods are required to derive the precise prognostic significance of p27Kip1 expression in breast cancer patients.
Biomarkers, Tumor ; analysis ; Breast Neoplasms ; diagnosis ; genetics ; metabolism ; pathology ; Carcinoma ; diagnosis ; genetics ; metabolism ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; metabolism ; Disease-Free Survival ; Female ; Gene Expression Regulation, Neoplastic ; genetics ; Humans ; Lymphatic Metastasis ; diagnosis ; physiopathology ; Neoplasm Staging ; methods ; Prognosis

Objective To construct a chimeric protein of Plasmodium falciparum pre erythrocyte stage (named as PfCP 4). Methods Thrombospondin related anonymous protein (TRAP) and circumsporozoite protein (CSP) of Plasmodium falciparum have been considered important candidates for pre erythrocytic malaria vaccine. The sequences of ectodomain of TRAP (aa: 26-330) and (NANP) 19 repeat region and entire carboxy terminus of CSP were fused to generate the PfCP 4 via a hinge consisting of Gly Pro Gly. The 1577 bp sequence of PfCP 4 was synthesized by asymmetric PCR based method and the synthetic gene was inserted into pQE. The resulting plasmid was transformed into E. coli SG13009 for inducible expression with IPTG. The expression product was detected by Western blotting. Results The result of Western blotting showed that the entire PfCP 4 recombinant protein was produced under IPTG induction whereas no product was detected in the cell without induction. The molecule weight of the protein was 57 kDa which was identical to the expected size, and the product was recognized by polyclonal antibodies against CSP protein. Conclusion A chimeric protein of Plasmodium falciparum pre erythrocyte stage (named as PfCP 4) was constructed successfully.