A highly sensitive immuno-PCR assay based on sandwich ELISA and PCR was developed to detect the circulating antigen in trichinellosis. Antigens were purified from the muscle larvae of Trichinella spiralis, and the myeloma cells were fused with spenocytes immunized with T. spiralis antigens to product the specific monoclonal antibodies. Indirect ELISA was used to select the antibody-secreting hybrodoma cells. By this method of procedure, monoclonal antibody F4C6 against the T. spiralis ES antigen was obtained, which was used as the indicator antibody, while the rabbit polyclonal antibodies against T. spiralis were to be used as capturing antibodies. The plasmid Bluecript Ⅱ KS was amplified by PCR with biotin-labeled primer M13-20, and thus the biotin-labeled DNA was obtained. Both the second antibody and DNA labeled with biotin were to be linked with 100 ng/ml avidin. The whole procedures of assay consisted of two steps, in which the circulating antigens were captured by monoclonal antibody through sandwich ELISA in the first step, and the DNA linked by monoclonal antibody was amplified by PCR in the second step. The sensitivity of this method was compared with that of the ELISA assay. It was found that the measuring ranges to detect the circulating antigens in trichinellosis were 50 pg/L to 0.005 pg/L for the immuno-PCR assay, and 5 μg/L to 0.05 μg/L for ELISA assay, the former was quite higher than that of the latter. It is evident that this method is highly sensitive for the detection of circulating antigens in trichinellosis.

Objective To intervene hand hygiene(HH)of health care workers (HCWs)in an intensive care unit(ICU)of a hospital,and evaluate HH compliance of HCWs and influencing factors.Methods 37 HCWs from an ICU of a teaching hospital were chosen as studied subjects,HH status before and after intervention was observed and analyzed by direct and indirect methods.Results In the first monitoring stage,HH rate in baseline and post-intervention survey stage was signifi-cantly different (46.96% vs 63.30%,χ2 =7.344,P =0.007);in the second monitoring stage,HH compliance rate in-creased from 57.85% to 88.58%.HH compliance rate was positively correlated with consumption of HH product per bed-day(r=0.725,P <0.001),but negatively correlated with incidence of healthcare-associated infection(HAI)and case rate of HAI(r=-0.983,-0.990,respectively,both P <0.001),HH rate to alcohol-based hand sanitizers was positively correlated with consumption of HH product per bed-day (r=0.391,P <0.001 ).Conclusion Long-term monitoring and intervention of HH compliance is necessary,combination of direct observation and indirect assessment of HH product con-sumption is helpful for effective monitoring and intervention of HH compliance among HCWs.

OBJECTIVEA novel method of Nano-Immunomagnetic Separation (Nano-IMS) plus Real-time PCR was established for detecting Vibrio cholerae.

METHODSThe Nano-Immunomagnetic Beads were created by using the monoclonal antibody of Vibrio cholerae, which was named Nano-IMB-Vc. Nano-IMB-Vc has specific adsorption of Vibrio cholerae, combined with Real-time PCR technology, a method for rapid detection of Vibrio cholerae was established. The capture specificity of Nano-IMB-Vc was tested by using 15 bacteria strains. The specificity of Real-time PCR method was tested by using 102 targets and 101 non-targets bacteria strains. The sensitivity of Nano-IMS plus Real-time PCR were tested in pure culture and in artificial samples and compared with NMKL No.156.

RESULTSThe capture ratio of Nano-IMB-Vc was reached 70.2% at the level of 10(3) CFU/ml. In pure culture, the sensitivity of Nano-IMS plus Real-time PCR was reached at 5.4×10(2) CFU/ml. The specific of Real-time PCR method was tested by using 102 targets and 101 non-targets bacteria. The results showed that 102 strains of Vibrio cholerae test results were all positive, and the rest of the 101 strains of non-target bacteria test results were negative. No cross-reaction was founded. Add 1 CFU vibrio cholerae per 25 g sample, it could be detect with Nano-IMS plus Real-time PCR method after 8 hours enrichment.

CONCLUSIONSThe Nano-IMS plus Real-time PCR method of Vibrio cholerae established in this study has good specificity and sensitivity, which could be applied to the rapid detection of Vibrio cholerae.

Food Microbiology ; methods ; Immunomagnetic Separation ; methods ; Nanotechnology ; Real-Time Polymerase Chain Reaction ; methods ; Seafood ; analysis ; microbiology ; Vibrio cholerae ; genetics ; isolation & purification ; Vibrio parahaemolyticus ; genetics ; isolation & purification

Objective : To study and compare the recurrence rate of wisdom tooth pericoronitis and impacted tooth types of Yunnan college ethnic student. Methods : 549 cases of pericoronitis of wisdom tooth from Yunnan college ethnic students were randomly selected by recording clinical symptom, recurrence situations and impacted tooth types by professional dentists, then clinical data was statistical analyzed. Results: Most of pericoronitis (58.65%) was caused by vertical impaction of mandibular wisdom tooth. There was no significant difference (P>0.05) between Han ethnic groups and Minority ethnic groups of impacted tooth types. But there was significant difference (P<0.01) between Han ethnic groups and Minority ethnic groups of pericoronitis of wisdom tooth and recurrence rate. Conclusions There was higher rate of incidence and recurrence rate of pericoronitis of minority students.

Objective To investigate the mental health status and its influencing factors among elderly hypertensive patients from Rural Areas of Chuxiong and Honghe Prefecture in Yunnan.Methods Multi-stage random sampling method was adopted to select elderly hypertensive patients from rural Yi ethnic areas in Yunnan.Questionnaires were used to collect their basic information and mental health status.Multivariate logistic regression was performed to explore the influencing factors of mental health among the elderly hypertensives.Results 21.82%(209/958)of elderly people with hypertension have poor mental health status in Chuxiong and Honghe Prefecture,Yunnan.Age of 80-89 years(OR = 2.395,P<0.05)and over 90 years(OR = 3.293,P<0.05),as well as physical disability(OR = 2.037,P<0.05),were risk factors for poor mental health.Compared with those who rated their economic situation as very difficult,rating as somewhat difficult(OR = 0.490,P<0.05),moderate(OR = 0.632,P<0.05)and relatively affluent(OR = 0.344,P<0.05),having a spouse(OR = 0.655,P<0.05),received full concern from the offspring(OR = 0.411,P<0.05)and maintain good relationships with offspring(OR = 0.339,P<0.05)were protective factors.Conclusions The mental health status of elderly people with hypertension is relatively poor in rural areas of Chuxiong and Honghe Prefecture in Yunnan Province.Special attention should be paid to the mental health of older and physically disabled elderly hypertensives.Economic and mental support from children was crucially important in improving the mental health of elderly hypertensive patients in rural areas of Chuxiong and Honghe Prefecture in Yunnan Province.

In order to further promote the reform of medical service price and strengthen the macro management of medical service price, the author constructed a calculation method for price adjustment space of medical service. This study was centered on two parameters of the historical base and the growth coefficient, to calculate the total amountof price adjustment. The historical base used the total revenue of medical services in the region from the previous year, and the growth coefficient was measured by two methods, the comprehensive indicator method and the consumer price index (CPI) reference method.Taking Suzhou, a national pilot city, as an example, the historical base in 2022 was 18 754 million yuan. By using the comprehensive indicator method, the annual growth coefficient was calculated to be 2.38%, and adjustment space of medical service price was 446.35 million yuan; According to the CPI reference method, the growth coefficient of Suzhou was 2.10%, and adjustment space of medical service price was 393.83 million yuan.The two methods for calculating the total amount each have their own advantages and disadvantages, and need to be further optimized and improved by drawing on the practical experience of reform in each pilot city.

Objective:To design a set of evaluation index system for the medical service price reform mechanism, so as to provide reference for the evaluation of deepening the medical service price reform mechanism.Methods:On the basis of searching literature of medical service prices reformfrom August 2021 to March 2023, policy logic analysis and expert consultation were used to construct evaluation indexes for the medical service price reform mechanism, set evaluation index thresholds, and assign scores to index thresholds in different grades.Results:The evaluationindex system of medical service price reform mechanism included 5 level-1 indexes, 12 level-2 indexes and 35 level-3 indexes. After scoring, the scores of 5 level-1 indexes were 12, 24, 35, 21, and 8, respectively.Based on the percentile scoring method, the evaluation criteria were categorized into five grades: premium, excellent, good, average and failure.Conclusions:This study constructed a set of evaluation index system for the mechanism of medical service price reform, which had strong scientific and operability. However, due to the fact that medical service price reform in the new era is still in its initial stage, the index system need to be further optimized drawing on the practical experience of reforms in pilot cities.

Objective Using network pharmacology to explore the mechanism of action of tripterygium wilfordii-white peony in the treatment of rheumatoid arthritis(RA)and validating it using an adjuvant arthritis rat model.Methods Obtain the effective ingredients of tripterygium wilfordii and white peony through TCMSP database and predict the action targets;Obtain the disease targets of RA using DisGeNET and GeneCards online database;Draw the network of"tripterygium wilfordii-white peony-active ingredient-RA"and screen the core effective components using the software Cytoscape 3.9.1;Construct the protein-protein interaction(PPI)network using String database and screen the core targets using the CytoNCA plug-in,using the Metascape database for gene enrichment analysis of common targets,and using Auto Dock Tools and Pymol software for molecular docking of core active ingredients and targets.Enzyme linked immunosorbent assay(ELISA)detection of serum tumor necrosis factor-α(TNF-α),interleukin(IL)-4,IL-6,and IL-17 levels in adjuvant arthritis rats,Western blot was used to detect the expression of phosphorylated phosphatidylinositol 3-hydroxy kinase(p-PI3K)and phosphorylated AKT protein(p-AKT),validate the results of network pharmacology analysis.Results There were a total of 61 effective ingredients in the treatment of RA with tripterygium wilfordii-white peony,with 116 targets and 191 signaling pathways involved.Animal experiments have shown that compared with the model group rats,the serum cytokine TNF-α,IL-6 and IL-17 of rats in tripterygium wilfordii-white peony group and tripterygium wilfordii extract group were significantly decreased(P<0.05),IL-4 significantly increased(P<0.05);The expression of p-PI3K and p-AKT in synovial tissue was significantly reduced(P<0.05);Compared with rats in tripterygium wilfordii extract group,the TNF-α,IL-6,IL-17,p-PI3K and p-AKT of rats in tripterygium wilfordii-white peony group were significantly reduced(P<0.05),while IL-4 was significantly increased(P<0.05).Conclusion Tripterygium wilfordii-white peony can inhibit the overexpression of phosphatidylinositol 3-hydroxy kinase(PI3K)/protein kinase B(AKT)signaling pathway,regulate cytokine release,exert anti-inflammatory effects,and ultimately treat RA through multiple active ingredients and targets.

OBJECTIVETo identify the causative mutations in two Chinese Han families featuring triphalangeal thumbs (TPT) and preaxial polydactyly (PPD).

METHODSBlood samples were collected from 9 members (2 affected) from family 1 and 14 members (7 affected) from family 2. After genomic DNA was extracted, the ZPA regulatory sequence (ZRS) region was analyzed with real-time quantitative PCR (qPCR) and Sanger sequencing. For family 1, haplotypes compassing the ZRS were also analyzed with short tandem repeats (STR) and single nucleotide changes.

RESULTSNo copy number mutation around the ZRS region was found in both families. Two heterogeneous mutations in the ZRS (406A>G and 105C>G) were found to co-segregate with the TPT/PPD malformation in family 1 and 2, respectively. Neither mutation was detected in 200 healthy individuals. Haplotype analysis and Sanger sequencing of family 1 indicated that the first TPT/PPD patient in the family was both germline and somatic mosaic for the 406A>G mutation.

CONCLUSIONTwo pathogenic ZRS mutations, 105C>G and 406A>G, have been identified in two Chinese Han families with TPT/PPD, among which the 406A>G mutation was de novo.

Asian Continental Ancestry Group ; genetics ; Female ; Hand Deformities, Congenital ; genetics ; Haplotypes ; Humans ; Male ; Mutation ; Polydactyly ; genetics ; Regulatory Sequences, Nucleic Acid ; Thumb ; abnormalities

Objective To identify the pathogenic variants in 110 patients with essential tremor(ET).Methods Clinical data and peripheral blood samples of ET patients were collected from the Department of Neurology of Peking Union Medical College Hospital and then the genomic DNA was extracted.Dynamic mutation detection of NOTCH2NLC was performed in patients with essential tremor by triplet repeat primed PCR(TP-PCR).Since ET is as-sociated with multiple mechanisms of neuro-degeneration,the next generation sequencing(NGS)panel targeting neu-rodegenerative associating genes were performed to check pathogenic variants in additional genes.Results A total of 110 ET patients and 187 matched control individuals were recruited.The age of onset in the current ET group was(36.30±17.64)years,and 74.8%patients had a family history.No abnormal trinucleotide repeat expansion in NOTCH2NLC was identified.The repeat number of(GGC)n lied within normal ranges between 10－47(average 18.6±5.4).Variants burden analysis showed association of ET with PLA2G6.Three rare variants in four patients in PLA2G6 were identified with unknown significance.Conclusions Dynamic mutations of NOTCH2NLC are uncom-mon in ET patients and that suggests need of more researches for further exploring the genetic mechanism of ET.