Objective:To explore the deubiquitination modification of the deubiquitinase BRCC3 in rats with neuropathic pain (NPP) treated with meridian tuina of Zhuang medicine and its analgesic molecular mechanism.Methods:Rats were divided into normal group, sham-operation group, model group, sham-tuina group, and meridian tuina of Zhuang medicine group using a random number table method, with 9 rats in each group. Except for the normal and sham operation groups, spinal nerve ligation models were prepared in all other groups. On the first day after surgery, intervention was carried out on the meridian tuina of Zhuang medicine and sham-tuina groups for 14 days, while the other three groups were not intervened; the paw withdrawal mechanical threshold (PWMT) and paw withdrawal thermal latency (PWTL) of each group of rats were measured at 0 days before operation, 1, 7, and 14 days after operation. Western blot was used to detect the expressions of BRCC3 and NLRP3 proteins in spinal cord tissue, and ELISA was used to detect the level of IL-1β in serum.Results:On postoperative 7 and 14 d, compared with the model group, the meridian tuina of Zhuang medicine group showed an increase in PWMT and PWTL, a decrease in NLPR3 and BRCC3 expression in spinal cord tissue, and a decrease in serum IL-1β levels ( P<0.05). Compared with the sham-tuina group, the meridian tuina of Zhuang medicine group showed an increase in PWMT and PWTL, a decrease in NLRP3 protein expression in spinal cord tissue, and a decrease in IL-1β levels in serum ( P<0.05). Conclusion:Meridian tuina of Zhuang medicine can alleviate pain sensitivity in SNL model rats, and its mechanism is related to the inhibition of the expression of deubiquitinase BRCC3 by meridian massage of Zhuang medicine, which increases the ubiquitination level of NLRP3 and hinders its activation, thereby blocking the immune inflammatory response mediated by inflammatory factors.

ObjectiveTo explore the correlation between serum albumin levels and coronary artery calcification （CAC） in patients with early-stage chronic kidney disease （CKD）， as well as the value of serum albumin levels in predicting the incidence and severity of CAC. MethodsThe study included 391 early-stage CKD patients who underwent coronary computed tomography angiography （CTA） at Sun Yat-sen Memorial Hospital of Sun Yat-sen University between January 2019 and December 2022. Demographic and biochemistry data， as well as the coronary CTA results， were collected. Based on the coronary artery calcification score （CACS）， all patients were divided into non-CAC group （CACS=0， n=184） and CAC group （CACS>0， n=207）. All patients were further divided into 3 groups based on the serum albumin levels： group A （serum albumin levels<35 g/L， n=30）， group B （35 g/L≤ serum albumin levels< 40 g/L， n=198） and group C （serum albumin levels≥ 40 g/L， n=163）. Univariate and multivariate binary logistic regression analyses were conducted to investigate the association between serum albumin levels and CAC in early-stage CKD patients. Differences in CAC among groups were analyzed by using post-hoc multiple comparisons and ordinal logistic regression model analysis. ResultsPatients with CAC had significantly lower serum albumin levels than those without CAC （P<0.05）. There was a negative correlation between serum albumin levels and CACS in early-stage CKD patients （P<0.01）， as serum albumin decreased in levels， CAC increased in severity. ConclusionsOur study shows that early-stage CKD patients with lower serum albumin levels have a higher incidence of CAC. Low serum albumin level is an independent risk factor for CAC progression.

【Objective 】 To explore the influencing factors of Yang deficiency constitution in traditional Chinese medicine (TCM) from the perspective of mathematics with the use of calculation formulas, so as to protect patients from getting diseases caused by Yang deficiency constitution and provide suggestions for TCM disease prevention. 【Methods】  Based on the classification and determination criteria of TCM constitution implemented by China Association of Chinese Medicine, data for 24 solar terms from May 5, 2020(Start of Summer) to April 20, 2021 (Grain Rain) for the identification of Yang deficiency were collected by mobile constitution identification system. The grey correlation analysis method was used to determine the grey correlation degree of the factors influencing Yang deficiency constitution. In addition, a random forest model was constructed for the verification of the results from the grey correlation analysis, and for the evaluation of correlation degree between Yang deficiency constitution and its influencing factors. 【Results】  A total of 16 259 sets of data were collected from healthy or sub-healthy individuals aged from 18 to 60 years living in the central and northeastern parts of Sichuan Province(China) for the identification of TCM constitutions. After screening and preprocessing, a total of 544 sets of data for the identification of Yang deficiency constitution, involving 18 aspects of factors influencing Yang deficiency constitution. The results of the grey correlation analysis showed that there were 12 influencing factors whose grey correlation degree with Yang deficiency constitution was greater than 0.6. The accuracy of these 12 influencing factors with the training set and validation set of the Yang deficiency constitution random forest model were 98.39% and 93.12%, respectively. 【Conclusion】  In the sample data selected in this paper, grey correlation analysis is the appropriate technology to analyze the influencing factors of Yang deficiency constitution. It provides a new idea and a new methodological reference for the research and analysis of the influencing factors of TCM constitution.

Objective To study the influencing factors of blood stasis constitution and provide a basis for treating blood stasis-related diseases by traditional Chinese medicine (TCM) constitution identification. Methods Data were collected using the self-developed TCM constitution identification platform based on B/S model by the project team. The obtained data were divided into blood stasis constitution and normal constitution groups. The differences of the categorical type influencing factors (gender, birth mode, feeding mode within four months of birth, family history, marital status, eating habits, sleeping habits, exercise habits, emotional state, stress situation, and living environment) and the quantitative type influencing factors (sleep time, age, and mother's age at birth) on the constitution of the two groups were analyzed. In the single-factor analysis, the Pearson's chi-square test was selected for the categorical variable, and the independent sample t test and Mann-Whitney U nonparametric test were selected for the quantitative variables according to whether they conformed to the positive-terrestrial distribution; the binary logistic stepwise regression method was selected for the multi-factor analysis. Results The data of 318 cases were collected from the TCM composition identification platform, and 159 cases of blood stasis constitution were used as the experimental group and 159 cases of normal constitution were used as the control group. The Pearson's chi-square test yielded significant differences (P < 0.05) in the effects of gender, pressure situation, family history, living environment, emotional state, exercise habits, and dietary habits on blood stasis constitution. The independent samples t test yielded differences in sleep duration between the blood stasis constitution and normal constitution populations (P < 0.05), which meant sleep duration of the blood stasis constitution population was less than that of the normal constitution population. The Mann-Whitney U nonparametric test results accepted the original hypothesis that there was no difference in the distribution of age and mother’s age at birth across constitution types (P > 0.05). Binary logistic regression analysis showed that gender, family history, marital status, living environment, exercise habits, and emotional state were risk factors for blood stasis constitution (P < 0.05). Conclusion Gender, family history, living environment, emotional state, and exercise habits were significant influencing factors of blood stasis constitution. Blood stasis constitution populations can pay more attention to these influencing factors in their daily life for the prevention and reconciliation of blood stasis constitution.

Objective To investigate the nutrient contents of various grains in Hubei Province, and to provide a scientific basis for a balanced diet. Methods Various grains (except potatoes and beans) on the market in Hubei Province were collected. The samples were prepared by peeling and milling, or directly milling, and the nutrient contents were determined according to the national standard methods. With reference to the American nutrient profiling model, protein, dietary fiber, vitamin C, vitamin A, vitamin E, potassium, magnesium, iron and calcium, the nine nutrients were selected as recommended nutrients, and sodium, added sugars and fat were restricted nutrients. On the basis of 100 g for calculation, a nutrient-rich food model (Nutrient-rich foods, NRF_9.3) was established and applied to evaluate the grains. Results The evaluation results showed that wheat and buckwheat grains contained a higher recommended nutrient content than restricted nutrient content, and had higher nutritional value. Among them, quinoa (black) had the highest NRF index of 102.4, indicating the most nutritional value. After peeling, the loss rate of nutrient value (NRF_9.3 index) of various grains was 38.73% to 65.00%. Conclusion It is recommended that people should try to choose whole grains when purchasing grain products.

OBJECTIVE: To detect mutation of LBR gene in a pedigree affected with Pelger-Huёt anomaly (PHA) and to explore its clinical characteristics. METHODS: Genomic DNA was extracted from the pedigree and healthy controls. The 14 exons of the LBR gene were subjected to PCR amplification and Sanger sequencing. Suspected mutations were verified in other family members and 100 healthy controls. Polyphen-2 and SIFT software were used to predict the effect of the mutation, and Swiss-model software was used to simulate the protein structure. RESULTS: Three patients were found to carry a c.893G>A mutation in exon 8 of the LBR gene, which resulted in substitution of the 298th amino acid residue glycine by glutamic acid (p.Gly298Glu). The same mutation was not found in healthy family members and 100 healthy controls. The mutation was predicted to be damaging. Bioinformatic simulation showed the mutation has altered the 3D structure of the LBR protein. CONCLUSION The c.893G>A (p.Gly298Glu) mutation in the LBR gene probably underlies the PHA in this pedigree and has enriched the spectrum of LBR gene mutations.
Case-Control Studies ; DNA Mutational Analysis ; Exons ; Humans ; Mutation ; Pedigree ; Pelger-Huet Anomaly ; genetics ; Polymerase Chain Reaction ; Receptors, Cytoplasmic and Nuclear ; genetics

Objective: To detect mutation of LBR gene in a pedigree affected with Pelger-Huёt anomaly (PHA) and to explore its clinical characteristics. Methods: Genomic DNA was extracted from the pedigree and healthy controls. The 14 exons of the LBR gene were subjected to PCR amplification and Sanger sequencing. Suspected mutations were verified in other family members and 100 healthy controls. Polyphen-2 and SIFT software were used to predict the effect of the mutation, and Swiss-model software was used to simulate the protein structure. Results: Three patients were found to carry a c. 893G>A mutation in exon 8 of the LBR gene, which resulted in substitution of the 298th amino acid residue glycine by glutamic acid (p.Gly298Glu). The same mutation was not found in healthy family members and 100 healthy controls. The mutation was predicted to be damaging. Bioinformatic simulation showed the mutation has altered the 3D structure of the LBR protein. Conclusion The c. 893G>A (p.Gly298Glu) mutation in the LBR gene probably underlies the PHA in this pedigree and has enriched the spectrum of LBR gene mutations.

Objective To establish genotyping methods for vitamin K epoxide reductase complex subunit 1 (VKORC1)and cytochrome P450 2C9(CYP2C9)based on pyrosequencing technique to detection of warfarin metabolizing enzyme related gene polymorphisms.Methods A total of 50 peripheral blood samples from healthy adults were collected and the whole blood genomic DNA was extracted.A set of biotin-labeled amplifi-cation primers and sequencing primers were designed respectively for three SNP sites:VKORC1 -1639 G>A,CYP2C9 430C> T and CYP2C9 1075A>C.After PCR amplification of the samples,pyrophosphoric acid se-quencing was conducted.And then the signal peaks form were combined to analyze and determine each sample genotype.Genotyping results were verified by Sanger sequencing,and the consistency of the two sequencing methods was compared.Results Genotypes of the three SNPs can be clearly determined according to the ba-ses and height of the signal peaks.Among the 50 samples,there were 41 AA and nine AG for VKORC1 -1639G>A,accounting for 82% and 12% respectively,and there were 45 *1/*1,five *1/*3 for CYP2C9, accounting for 90% and 10% respectively,no CYP2C9*2 allele detected.Genotype results detected by pyrose-quencing and Sanger sequencing were consistent with each other.Conclusion In SNP genotyping,Pyrose-quencing has the advantages of convenience,time-saving,cheap with accurate and reliable results,which can quickly determine the genotypes of CYP2C9 and VKORC1.

Objective To investigate the methods and internal quality control ( IQC ) leucorrhea routine examinationin clinical laboratories of medical institutions in Guizhou Province.Methods In 2009, 97 clinical laboratories were randomly selected for the first investigation.At the same time, staffs in theinvestigated laboratories were educated on the importance of IQC.The second investigation of the same items was carried out in 2011 inthe same laboratories.The results of the two investigations were analyzed byChi-square test.Results 2009 and 2011 numbers of laboratories thoseonly used normal saline suspension method for leucorrhea examination were 17and 16 (χ2 =0.037, P >0.05 ) respectively, used bothnormal saline and 10%KOH suspension methodswere 16and 2(χ2 =12.003,P<0.01), used staining method were 64and 79(χ2 =5.488,P<0.05), both used suspension and staining methods were 60and 73(χ2 =4.041, P<0.05), used normal salinesuspension method combined with Wright stain and Gram staining methods were3and 28(χ2 =23.996,P<0.01) respectively.Numbers of Laboratoriespracticing IQC were 2and 88in 2009 and 2011 respectivly(χ2 =153.293,P <0.01).Conclusions Currently, the most common used method for leucorrhea routine examination is suspension.Through the investigations and education, the quality ofleucorrhea routine examination was improved in Guizhou Province.

OBJECTIVE To evaluate the clinical application of T-cell based IFN-? release assays(IGRA) for the rapid diagnosis of active tuberculosis.METHODS IFN-? and HIV antibody were detected by using ELISA.Antibody to Mycobacterium tuberculosis was detected by colloidal gold.At the same time,the M.tuberculosis DNA loads were examined by FQ-PCR.Statistical analysis were performed to analyze the correlation of IFN-? with M.tuberculosis antibody and DNA,respectively.RESULTS The sensitivity of TB-IGRA was 90.24%,specificity was 93.34%;the positive rate of TB-IGRA in 82 tuberculosis patients was higher than from sputum smear(64.63%),TB-PCR(76.83%) and tuberculosis antibody(40.24%).CONCLUSIONS As a replacement of TB-PCR,IFN-? can be used as a valued index to evaluate tuberculosis infectin.