Objective To establish a rapid,sensitive,and specific multiplex PCR detection method for the simultaneous detection of Cryptosporidium,Eimeria,and bovine parvovirus.Methods Specific primers targeting the SSU rRNA genes of Cryptosporidium and Eimeria,as well as the VP2 gene of bovine parvovirus were designed and the corresponding recombinant plasmid standards were constructed.To establish the multiplex PCR method,the reaction conditions were optimized using temperature gradient PCR and single-variable control methods.The sensitivity,specificity,reproducibility,and clinical application of the protocol were evaluated.Results The optimal annealing temperature was found to be 60.5℃,and the forward and reverse primer concentrations were determined to be 0.2 μmol/L for Eimeria,and 0.4 μmol/L for Cryptosporidium and bovine parvovirus.The assay demonstrated high sensitivity,with detection limits of 243,260,and 3 110 copies for the recombinant plasmid standards of Cryptosporidium,Eimeria,and bovine parvovirus,respectively.Specificity testing showed no cross-reactivity with ten common bovine pathogens,including Salmonella,bovine viral diarrhea virus,and bovine rotavirus.Consistent intra-and inter-batch results confirmed the strong reproducibility of the method.Clinical application to 81 diarrhea samples from various regions in the Ganzi Prefecture,Sichuan,revealed positivity rates of 18.52%(15/81)for Cryptosporidium,34.57%(28/81)for Eimeria,and 18.52%(15/81)forbovineparvovirus,withamixedinfectionrateof3.7%(3/81).Conclusions Themultiplex PCR method established in this study offers a reliable tool for differential diagnosis and epidemiological investigation of the three common diarrheal pathogens in yaks.

Objective To understand the willingness of general practitioner(GP) to enhance working competence in community healthcare centers in Shanghai and provide a basis for the competence training of GPs in community healthcare centers. Methods In August 2023,GPs were selected from some community healthcare centers in Shanghai and their willingness to enhance working competence were studied by a questionnaire survey.The survey included 39 secondary indicators in three dimensions:general practice theory,skills,and humanity. Results A total of 1 192 GPs completed the questionnaire,with an effective rate of 100%.The total score of GPs' willingness to enhance their working competence was 258.45±80.93,and the mean score of the three dimensions was 6.63±2.08.The score for the general practice theory was the highest (6.92±1.95),while that for general practice humanity was the lowest (6.44±2.34) among the three dimensions.The score of willingness to enhance working efficiency differed across different age ranges (P<0.001),professional titles (P<0.001),years of work (P<0.001),and educational backgrounds of GPs (P=0.039).Those with the age younger than 30 years old,junior professional titles,less than 5 years of work experience,and a college degree or below had the highest willingness score to enhance their working competence.Among the top three secondary indicators of willingness score in each dimension,the top three methods of working competence enhancement were community general practice and specialized healthcare services combined with outpatient learning,flexible further training,and continuing education courses.Conclusions There is an urgent need for young GPs in community healthcare centers in Shanghai to enhance their working competence.Targeted enhancement plans can be provided to different groups of GPs with different characteristics through community general practice and specialized healthcare services combined with outpatient learning,flexible further training,and continuing education courses,which can further enhance the ability and quality of the GP team.
Humans ; China ; General Practitioners/psychology* ; Surveys and Questionnaires ; Community Health Centers ; Clinical Competence ; Female ; Adult ; Male ; Attitude of Health Personnel ; Middle Aged

OBJECTIVE: To evaluate the dynamic changes of glucocorticoid (GC) dose and the feasibility of GC discontinuation in rheumatoid arthritis (RA) patients under the background of Chinese medicine (CM). METHODS: This multicenter retrospective cohort study included 1,196 RA patients enrolled in the China Rheumatoid Arthritis Registry of Patients with Chinese Medicine (CERTAIN) from September 1, 2019 to December 4, 2023, who initiated GC therapy. Participants were divided into the Western medicine (WM) and integrative medicine (IM, combination of CM and WM) groups based on medication regimen. Follow-up was performed at least every 3 months to assess dynamic changes in GC dose. Changes in GC dose were analyzed by generalized estimator equation, the probability of GC discontinuation was assessed using Kaplan-Meier curve, and predictors of GC discontinuation were analyzed by Cox regression. Patients with <12 months of follow-up were excluded for the sensitivity analysis. RESULTS: Among 1,196 patients (85.4% female; median age 56.4 years), 880 (73.6%) received IM. Over a median 12-month follow-up, 34.3% (410 cases) discontinued GC, with significantly higher rates in the IM group (40.8% vs. 16.1% in WM; P<0.05). GC dose declined progressively, with IM patients demonstrating faster reductions (median 3.75 mg vs. 5.00 mg in WM at 12 months; P<0.05). Multivariate Cox analysis identified age <60 years [P<0.001, hazard ratios (HR)=2.142, 95% confidence interval (CI): 1.523-3.012], IM therapy (P=0.001, HR=2.175, 95% CI: 1.369-3.456), baseline GC dose ⩽7.5 mg (P=0.003, HR=1.637, 95% CI: 1.177-2.275), and absence of non-steroidal anti-inflammatory drugs use (P=0.001, HR=2.546, 95% CI: 1.432-4.527) as significant predictors of GC discontinuation. Sensitivity analysis (545 cases) confirmed these findings. CONCLUSIONS RA patients receiving CM face difficulties in following guideline-recommended GC discontinuation protocols. IM can promote GC discontinuation and is a promising strategy to reduce GC dependency in RA management. (Trial registration: ClinicalTrials.gov, No. NCT05219214).
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Arthritis, Rheumatoid/drug therapy* ; Glucocorticoids/therapeutic use* ; Medicine, Chinese Traditional ; Retrospective Studies

Objective To observe the efficacy and safety of sclerotherapy of pelvic lymphoid cysts with anhydrous ethanol and lauromacrogol.Methods Totally 98 patients with pelvic lymphatic cysts after resection of gynecological malignant tumors were retrospectively enrolled,including 50 cases underwent sclerotherapy with anhydrous ethanol(anhydrous ethanol group)and 48 cases with lauromacrogol(lauromacrogol group).The complications after sclerotherapy were recorded,and the effect was evaluated according to improvement of symptoms and results of ultrasound,CT or MRI 3 months after sclerotherapy.Results Sclerotherapy was successfully completed in both groups.After sclerotherapy,adverse reactions occurred in 20 cases in anhydrous ethanol group,including pain(n=5),drunk-like reaction(n=2),cyst infection and drug resistance(n=1),intestinal perforation(n=1)and decreased immunity(n=6),among which 2 were common terminology criteria for adverse events(CTCAE)grade 1 and 18 were CTCAE grade 2-3 adverse reactions.Meanwhile,minor pain occurred in 2 cases in lauromacrogol group,both were CTCAE grade 1 adverse reactions.The incidence of CTCAE grade 2-3 adverse reactions in anhydrous ethanol group was higher than in lauromacrogol group(36.00％[18/50]vs.0[0/48],x2=21.168,P＜0.001).All the above adverse reactions in 2 groups improved after symptomatic support treatment,and no other complication was observed.Three months after treatment,4 cases were cured,40 were improved and 6 were ineffective in anhydrous ethanol group,while 7 were cured,40 were improved and 1 was ineffective in lauromacrogol group.No significant difference of cyst control rate was found between groups(88.00％[44/50]vs.97.92％[47/48],x2=4.178,P=0.128).Conclusion The effects of sclerotherapy of pelvic lymphoid cysts with anhydrous ethanol and lauromacrogol were equivalent,while the latter was safer.

Objective:This study aims to analyze the policy tools used in China's diabetes prevention and control services and provide recommendations for improving the diabetes policy system in China. Methods:From the perspectives of the three levels of prevention and policy tools,a two-dimensional framework of"Policy Tool Types-Three Levels of Prevention (including service categories)"was constructed through content analysis and quantitative analysis methods. This framework was used to analyze the national-level diabetes prevention and control service policy entries. Results:A total of 88 policy documents related to diabetes prevention and control services were included in this study. There were 181 codes for the policy tool dimension. From the perspective of policy tools,mandatory policy tools were mainly used to ensure the implementation of services,with the proportions of mandatory,mixed,and voluntary policy tools being 74.6％,19.3％,and 6.1％ respectively,showing significant distribution differences. From the perspective of the three levels of prevention,strategies at all levels were covered broadly,with the proportions of the three levels of prevention being 38.7％,36.5％,and 24.9％ respectively,and internal sub-tools needing optimization. From the perspective of service categories,patient classification and follow-up management,and health education were the most common,while baseline disease surveys were less frequent. There was a lack of targeted policy tools for different types of prevention and control services. Conclusion:The diabetes policy tools exhibit structural imbalances,the use of internal sub-tools needs to be optimized,and there is a significant difference in the distribution of different prevention levels of diabetes. Enhance the precision of policy tools and strengthen the effective combination of different diabetes prevention and control service characteristics with policy tools;optimize the structure of policy tools and emphasize the coordinated operation of policy tools with the three levels of prevention strategies.

Objective To explore the effect of glioma stem cells with high expression of protein tyrosin phosphatase receptor type Z1 (PTPRZ1 )on the phenotypic polarization and phagocytosis of tumor-associated macrophages and its regulatory mechanism.Methods GSCs and non-stem tumor cells (NSTCs) were screened out from human glioblastoma (GBM) specimens using flow cytometry,and the PTPRZ1 expression in paired GSCs and NSTCs were detected.Human peripheral blood mononuclear cells (PBMC)-derived CD14+monocytes were exposed to the conditioned medium from glioma cells or recombinant chemokine C-C motif ligand 20 (CCL20)for TAM polarization.Stable PTPRZ1 knockout GSCs (PTPRZ1-KO GSCs) were constructed using CRISPR/Cas9. TAM phagocytosis to GSCs,NSTCs,PTPRZ1-Control GSCs (PTPRZ1-Ctrl GSCs)and PTPRZ1-KO GSCs and the expression of immunosuppressive phenotype (M2) polarization marker CD163 were examined using flow cytometry.Differentially expressed genes (DEGs ) between paired GSCs and NSTCs were determined using a bulk RNA-sequencing dataset (GSE54791 )from Gene Expression Omnibus (GEO).A gene set informing worse outcome of patients with GBM was generated using The Cancer Genome Atlas (TCGA)-GBM cohort.By intersecting the aforementioned gene set with the gene set that encodes for human membrance proteins,the PTPRZ1 gene is obtained.Gene set enrichment analysis (GSEA)was used for pathway enrichment analysis to compare the differentially regulated pathways between GBMs with high or low PTPRZ1 expression.Bulk RNA sequencing,qRT-PCR and Western blotting were used to identify the DEGs between PTPRZ1-KO GSCs and PTPRZ1-Ctrl GSCs.Results GSCs were more capable of escaping from TAM phagocytosis than NSTCs (P<0.05 )and had specifically up-regulated PTPRZ1 expression.PTPRZ1-KO significantly suppressed GSCs escaping from TAM phagocytosis (P<0.01 ). GBMs with high PTPRZ1 expression showed significant inhibition of pathways mediating phagocytosis (P<0.05).The expression of CCL20 as a M2 TAM polarization chemokine was significantly down-regulated in PTPRZ1-KO GSCs (P<0.05 ).Treatment with recombinant CCL20 up-regulated the expression of CD163 as a M2 TAM marker in TAM.Conclusion PTPRZ1+GSCs mediate M2 TAM polarization and inhibit TAM phagocytosis,which may be related to the up-regulation of CCL20 in PTPRZ1+GSCs.

Objective:This study aims to analyze the policy tools used in China's diabetes prevention and control services and provide recommendations for improving the diabetes policy system in China. Methods:From the perspectives of the three levels of prevention and policy tools,a two-dimensional framework of"Policy Tool Types-Three Levels of Prevention (including service categories)"was constructed through content analysis and quantitative analysis methods. This framework was used to analyze the national-level diabetes prevention and control service policy entries. Results:A total of 88 policy documents related to diabetes prevention and control services were included in this study. There were 181 codes for the policy tool dimension. From the perspective of policy tools,mandatory policy tools were mainly used to ensure the implementation of services,with the proportions of mandatory,mixed,and voluntary policy tools being 74.6％,19.3％,and 6.1％ respectively,showing significant distribution differences. From the perspective of the three levels of prevention,strategies at all levels were covered broadly,with the proportions of the three levels of prevention being 38.7％,36.5％,and 24.9％ respectively,and internal sub-tools needing optimization. From the perspective of service categories,patient classification and follow-up management,and health education were the most common,while baseline disease surveys were less frequent. There was a lack of targeted policy tools for different types of prevention and control services. Conclusion:The diabetes policy tools exhibit structural imbalances,the use of internal sub-tools needs to be optimized,and there is a significant difference in the distribution of different prevention levels of diabetes. Enhance the precision of policy tools and strengthen the effective combination of different diabetes prevention and control service characteristics with policy tools;optimize the structure of policy tools and emphasize the coordinated operation of policy tools with the three levels of prevention strategies.

Objective To observe the efficacy and safety of sclerotherapy of pelvic lymphoid cysts with anhydrous ethanol and lauromacrogol.Methods Totally 98 patients with pelvic lymphatic cysts after resection of gynecological malignant tumors were retrospectively enrolled,including 50 cases underwent sclerotherapy with anhydrous ethanol(anhydrous ethanol group)and 48 cases with lauromacrogol(lauromacrogol group).The complications after sclerotherapy were recorded,and the effect was evaluated according to improvement of symptoms and results of ultrasound,CT or MRI 3 months after sclerotherapy.Results Sclerotherapy was successfully completed in both groups.After sclerotherapy,adverse reactions occurred in 20 cases in anhydrous ethanol group,including pain(n=5),drunk-like reaction(n=2),cyst infection and drug resistance(n=1),intestinal perforation(n=1)and decreased immunity(n=6),among which 2 were common terminology criteria for adverse events(CTCAE)grade 1 and 18 were CTCAE grade 2-3 adverse reactions.Meanwhile,minor pain occurred in 2 cases in lauromacrogol group,both were CTCAE grade 1 adverse reactions.The incidence of CTCAE grade 2-3 adverse reactions in anhydrous ethanol group was higher than in lauromacrogol group(36.00％[18/50]vs.0[0/48],x2=21.168,P＜0.001).All the above adverse reactions in 2 groups improved after symptomatic support treatment,and no other complication was observed.Three months after treatment,4 cases were cured,40 were improved and 6 were ineffective in anhydrous ethanol group,while 7 were cured,40 were improved and 1 was ineffective in lauromacrogol group.No significant difference of cyst control rate was found between groups(88.00％[44/50]vs.97.92％[47/48],x2=4.178,P=0.128).Conclusion The effects of sclerotherapy of pelvic lymphoid cysts with anhydrous ethanol and lauromacrogol were equivalent,while the latter was safer.

Objective: To understand the basic characteristics of previously reported patients with hepatitis C and analyze the related factors affecting their antiviral treatment. Methods: A convenient sampling method was adopted. Patients who had been previously diagnosed with hepatitis C in the Wenshan Prefecture of Yunnan Province and Xuzhou City of Jiangsu Province were contacted by telephone for an interview study. The Andersen health service utilization behavior model and related literature were used to design the research framework for antiviral treatment in previously reported hepatitis C patients. A step-by-step multivariate regression analysis was used in previously reported hepatitis C patients treated with antiviral therapy. Results: A total of 483 hepatitis C patients, aged 51.73 ± 12.06 years, were investigated. The proportion of male, agricultural occupants who were registered permanent residents, farmers and migrant workers was 65.24%, 67.49%, and 58.18%, respectively. Han ethnicity (70.81%), married (77.02%), and junior high school and below educational level (82.61%) were the main ones. Multivariate logistic regression analysis results showed that married patients with hepatitis C (OR = 3.19, 95% CI: 1.93-5.25, compared with unmarried, divorced, and widowed patients) with high school education or above (OR = 2.54, 95% CI: 1.54-4.20, compared with patients with junior high school education or below) were more likely to receive antiviral treatment in the predisposition module. Patients with severe self-perceived hepatitis C in the need factor module (compared with patients with mild self-perceived disease, OR = 3.36, 95% CI: 2.09-5.40) were more likely to receive treatment. In the competency module, the family's per capita monthly income was more than 1,000 yuan (compared with patients with per capita monthly income below 1,000 yuan, OR = 1.59, 95% CI: 1.02-2.47), and the patients had a high level of awareness of hepatitis C knowledge (compared with patients with a low level of knowledge, OR = 1.54, 95% CI: 1.01-2.35), and the family members who knew the patient's infection status (compared with patients with an unknown infection status, OR = 4.59, 95% CI: 2.24-9.39) were more likely to receive antiviral treatment. Conclusion: Different income, educational, and marital statuses are related to antiviral treatment behavior in hepatitis C patients. Family support of hepatitis C patients receiving hepatitis C-related knowledge and their families knowing the infection status is more important in promoting the antiviral treatment of patients, suggesting that in the future, we should further strengthen the hepatitis C knowledge of hepatitis C patients, especially the family support of hepatitis C patients' families in treatment.
Humans ; Male ; Antiviral Agents/therapeutic use* ; China ; Hepatitis C/drug therapy* ; Hepacivirus ; Logistic Models

Humans ; Otosclerosis/surgery* ; Stapes ; Stapes Surgery/adverse effects* ; Arteries