BACKGROUND: Previous research demonstrated that a homozygous mutation of g.136372044G>A (S12N) in caspase recruitment domain family member 9 ( CARD9 ) is critical for producing Aspergillus fumigatus -induced ( Af -induced) T helper 2 (T H 2)-mediated responses in allergic bronchopulmonary aspergillosis (ABPA). However, it remains unclear whether the CARD9S12N mutation, especially the heterozygous occurrence, predisposes the host to ABPA. METHODS: A total of 61 ABPA patients and 264 controls (including 156 healthy controls and 108 asthma patients) were recruited for sequencing the CARD9 locus to clarify whether patients with this heterozygous single-nucleotide polymorphisms are predisposed to the development of ABPA. A series of in vivo and in vitro experiments, such as quantitative real-time polymerase chain reaction, flow cytometry, and RNA isolation and quantification, were used to illuminate the involved mechanism of the disease. RESULTS: The presence of the p.S12N mutation was associated with a significant risk of ABPA in ABPA patients when compared with healthy controls and asthma patients, regardless of Aspergillus sensitivity. Relative to healthy controls without relevant allergies, the mutation of p.S12N was associated with a significant risk of ABPA (OR: 2.69 and 4.17 for GA and AA genotypes, P = 0.003 and 0.029, respectively). Compared with patients with asthma, ABPA patients had a significantly higher heterozygous mutation (GA genotype), indicating that p.S12N might be a significant ABPA-susceptibility locus ( aspergillus sensitized asthma: OR: 3.02, P = 0.009; aspergillus unsensitized asthma: OR: 2.94, P = 0.005). The mutant allele was preferentially expressed in ABPA patients with heterozygous CARD9S12N , which contributes to its functional alterations to facilitate Af -induced T H 2-mediated ABPA development. In terms of mechanism, Card9 wild-type ( Card9WT ) expression levels decreased significantly due to Af -induced decay of its messenger RNA compared to the heterozygous Card9S12N . In addition, ABPA patients with heterozygous CARD9S12N had increased Af -induced interleukin-5 production. CONCLUSION Our study provides the genetic evidence showing that the heterozygous mutation of CARD9S12N , followed by allele expression imbalance of CARD9S12N , facilitates the development of ABPA.
Humans ; Aspergillosis, Allergic Bronchopulmonary/complications* ; Aspergillus fumigatus/genetics* ; Asthma/genetics* ; Aspergillus ; Mutation/genetics* ; CARD Signaling Adaptor Proteins/genetics*

Objective:To identify the transmission relationship between HIV infection cases the non-marital non-commercial heterosexual contact in Zhejiang Province.Methods:When HIV positive was informed during January 2020 to January 2022, the staff conducted an epidemiological investigation to collect cases information on sociodemographic characteristics, mobility information, past HIV testing history, high-risk sexual behaviors, sexual partners, and etcetera. At the same time, 6-8 ml of blood from the new diagnosis of people infected with HIV before antiviral treatment was collected to separate the bleeding plasma. pol gene was amplified by nucleic acid extraction and PCR, sequenced by Sequencer 5.0 software, and Cytoscape 3.6.0 software was used to draw HIV molecular transmission network. Results:From January 2020 to January 2022, 88 HIV infected individuals were found in Pujiang County, of which 74 were transmitted through heterosexual transmission, of which 31 were infected through non-marital non-commercial heterosexual contact. Preliminary case studies have found that three female cases have engaged in unprotected non-marital non-commercial heterosexual contact with one male case. Among the 4 infected individuals, 2 of their spouses tested positive for HIV antibodies. Molecular transmission network monitoring was carried out on 65 newly diagnosed cases of heterosexual transmission with acquired sequences, forming 9 transmission clusters. The largest cluster contained 10 cases. A total of 11 HIV-infected individuals were involved in this HIV cluster epidemic. They were 3 males and 8 females, all over 50 years old and were farmers or rural housewives. They were traced to 7 sexual partners (6 negatives of HIV, 1 undetected). Among the 18 respondents' sexual social network relationships, there were 6 couples, 8 permanent partners, and 3 temporary partners. Among 11 HIV infected individuals, there were 9 cases of non-marital non-commercial heterosexual transmission and 2 cases of intramarital transmission. The epidemiological association between 7 non-married non-commercial heterosexual partners and case 2 (56-year-old male farmer), 3 cases confirmed by epidemiological investigation and molecular transmission cluster results, 3 cases confirmed by molecular transmission cluster and epidemiological investigation results, and 1 case confirmed by epidemiological investigation results.Conclusions:The transmission mode of this cluster epidemic was to spread HIV through heterosexual sex with a male case as the core, then cause the transmission within marriage and between fixed sexual partners. The combination of epidemiological investigation and molecular transmission network traceability survey supports the conclusion of this study.

BACKGROUND: Regenerative medicine by using stem cells from dental pulp is promising for treating patients with critical limb ischemic (CLI). Here, we investigated the difference in the angiogenetic ability of stem cells from human exfoliated deciduous teeth (SHED) and human dental pulp stem cells (DPSC). METHODS: SHED and DPSC were harvested from dental pulp and analyzed in flow- cytometry for detecting the expression of surface markers. Levels of angiogenetic marker were examined by RT-PCR and Western-blot. Eighteen immunodeficient mice of critical limb ischemic model were divided into three groups: SHED, DPSC and saline, which was administered with SHED, DPSC or saline intramuscularly. Histological examination was performed to detect the regenerative results. RESULTS: A highly expression of CD146 was detected in SHED. Moreover, cells with negative expression of both CD146 and CD31 in SHED were more in comparison with those in DPSC. Expression of angiogenesis factors including CXCL12, CXCR4, Hif-1a, CD31, VEGF and bFGF were significant higher in SHED than DPSC by the RT-PCR and Western-Blot results. SHED induced more CD31 expression and less fibrous tissue formation in the critical limb ischemic model as compare with DPSC and saline. CONCLUSION Both SHED and DPSC possessed the ability of repairing CLI. With expressing more proangiogenesis factors, SHED may have the advantage of repairing CLI.

Objective: To analyze the characteristics and influencing factors of the newly diagnosed cases locally infected with HIV through sexual contact in Jinhua,so as to provide reference for AIDS prevention and control. Methods: An epidemiological survey was conducted among the HIV/AIDS cases diagnosed in 2017 in Jinhua to collect the information about demographic characteristics,local infection and sexual behaviors. The logistic regression model was used to analyze the related factors for local HIV infection. Results: A total of 438 HIV/AIDS cases infected through sexual contact were recruited,with 272（62.10%）cases infected in Jinhua. The proportion of local infection was 86.67%,79.47%,63.04%,69.09%,77.46% and 77.97%,respectively,among those people aged 60 years or over,permanent residents in Jinhua,employees / students,farmers,those who had lived in Jinhua for more than five years,and those who had a HIV testing in one year. The Results of the multivariate logistic regression analysis showed that heterosexual transmission cases who were permanent residents in Jinhua（OR=3.437,95%CI：1.250-9.451）,who had lived in Jinhua for more than five years（OR=3.609,95%CI：1.403-9.284）,who had commercial heterosexual behaviors in Jinhua（OR=5.463,95%CI：2.529-11.803）were more likely to be infected with HIV in Jinhua;homosexual transmission cases who were permanent residents in Jinhua（OR=4.812,95%CI：1.744-13.275）and who had non-commercial,temporary homosexual behaviors in Jinhua（OR=10.641,95%CI：4.369-25.916）were more likely to be infected with HIV in Jinhua. Conclusion Among the HIV/AIDS cases diagnosed in 2017 infected through sexual contact in Jinhua,having permanent residence,long-term residence,commercial heterosexual behaviors and non-commercial,temporary homosexual behaviors were risk factors for local infection.

Objective To establish diagnosis model and explore related metabolic pathways by analyzing the serum metabolic profile of patients with primary nephrotic syndrome (PNS) through metabolomics.Methods Thirty PNS patients hospitalized in Huai'an First People's Hospital between December 2010 and April 2012 were enrolled.High performance liquid chromatography-mass spectrometry (LC-MS) was employed to detect metabolites in the serum of 30 PNS patients and 30 healthy controls.Metabolic fingerprint profiling and multivariate pattern recognition analysis were combined to establish disease-specific metabolic diagnosis model,and metabolic pathway analysis was performed.Results PNS group and control group could be well separated by principal component analysis (PCA) model as well as partial least-squares discriminant analysis (PLS-DA) model with Q2 of 0.300.There was well interpretation in PLA-DA model (R2X=0.581,R2Y=0.452).Compared with healthy controls,PNS patients had decreased cholestane 3,7,12,15 alcohol,acyl glycerine,phytosphingosine and tryptophan,and increased sphingomyelin,arginine and glutamic acid (all VIP ＞ 1,P ＜ 0.05).The metabolic disorders pathways of PNS patients included sphingolipid metabolism,arginine and proline metabolism,linoleic acid metabolism and pyrimidine metabolism (all impact ＞0.10 and P ＜ 0.05).Conclusions Metabolomics combined with multivariate pattern recognition analysis may be a new tool for diagnosis and monitoring of PNS.

Objective The metabolomics methods were applied to discover the serum differences metabolites in patients with lupus nephritis (LN),and to establish the diagnostic model and study the metabolic pathways change due to the interference of LN and explore the pathogenesis of LN.Methods Positive and negative ions mode of ultra high performance liquid chromatography-high resolution mass spectrometry (HPLS-HRMS) were used to analyze the serum samples of 38 LN patients and 35 healthy controls,respectively.Both univariate and multivariate data analysis were performed with SPSS software (V17.0,Chicago,USA) and SIMCA-P+ package (V12.0,Umetrics,Sweden),respectively.A supervised pattern recognition method,orthogonal partial least squares-discriminant analysis (OPLS-DA),was employed to study the metabolomics differences between control group and LN patients group.The metabolic pathways were analyzed by Metabo Analyst 3.0.Results LN patients and healthy controls could be well separated by OPLS-DA model Q2 of 0.583.Eighteen compounds (theophylline,oxidized glutathione,capric acid,norvaline,hippuric acid,sphingosine,sorbitol,cortisol,pyrrole-2-carboxylic acid,deoxycholic acid,tryptamine,5-aminolevulinic acid,glucose 6-phosphate,riboflavin,cortisone,taurine,creatinine,glycerophosphocholine) were initially identified as potential serum metabolic markers to distinguish LN patients from healthy people.Alternated metabolism pathways included citrate metabolism,pyrimidine metabolism and amino acid metabolism.Conclusion Through analysis of the metabolic pathways of different metabolites,the significant impact of LN on citrate metabolism,pyrimidine metabolism and amino acid metabolism pathways are identified.Some of the different metabolites are involved in the process of immunization.These results will benefit the early intervention and treatment of LN and the research of the mechanism of pathogenesis.

Objective To explore the effectiveness of mindfulness-based four-step approach to obsessive compulsive disorder. Methods A total of 82 patients with obsessive compulsive disorder were divided into control group and study group with 41 cases each by admission order. The patients in control group were given routine drug treatment and nursing, and the patients in study group were given mindfulness-based four-step approach based on control group. The indexes were evaluated by Yale-Brown Obsessive Compulsive Scale, Hamilton Depressive Scale, Hamilton Anxiety Scale, Short Form 36 questionnaire before and after intervention. Results After 12 weeks treatment, the score of Yale-Brown Obsessive Compulsive Scale, compulsive thinking, compulsive behavior were (6.16 ±4.38), (3.23 ±1.93), (2.98 ± 2.01) points in study group and (10.81 ± 3.26), (6.05 ± 2.02), (4.09 ± 2.15) points in control group, and there were significant differences(t=5.386, 6.384, 2.385, all P < 0.05). The score of anxiety and depress were (1.98 ± 1.91), (2.16 ± 1.38) points in study group, and (4.05 ± 2.01), (5.11 ± 2.26) points in control group, and there were significant differences(t=4.722, 7.045, all P<0.05). The score of total life quality, physiological function, physiological function, body pain, general health, vitality, social function, emotional function, mental health were (77.32 ± 6.26), (89.11 ± 14.27), (78.19 ± 26.36), (82.33 ± 25.22), (60.18 ± 20.39), (55.31 ± 19.68), (80.13 ± 25.76), (72.13 ± 20.16), (60.12 ± 20.88) points respectively in study group, and (71.43 ± 5.62), (82.19 ± 13.36), (55.11 ± 23.27), (70.18 ± 25.71), (49.01 ± 20.13), (45.22 ± 18.11), (62.19±25.63), (45.23±23.02), (44.16±21.08) points in control group, and there were significant differences (t=2.134-6.621, all P<0.05). Conclusions The mindfulness-based four-step approach intervention can obviously relieve anxiety and depressive negativity mood in obsessive compulsive disorder patients. It can improve patient′s obsessive compulsive symptom, enhance the patient′s quality of life. It has important significance for obsessive compulsive disorder patients during their recovering period.

〔Abstract〕 Taking Beijing University of Chinese Medicine as an example , the paper analyzes problems existing in teaching computer basic courses .It proposes to combine the teaching concept of flipped classroom with the case -based teaching method , reform the teach-ing contents, methods, processes and modes of computer basic courses in medical colleges and cultivate students ′abilities to analyze and solve problems , and analyzes the feasibility .

Objective To observe the change of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene expression during 3T3-L1 adipocyte differentiation as well as other reference gene expressions.Methods The mRNA expressions of several common reference genes were detected by real time-PCR on day 0,1,3,5,and 7 of 3T3-L1 adipocyte differentiation.Western blot was used to confirm the protein expressions of three common reference genes.Results (1) GAPDH and transferrin receptor(TFRC) mRNA expressions were significantly increased during adipocyte differentiation.GAPDH mRNA level was increased by 5.7,7.6,22.0,and 24.5 folds on day 1,3,5,and 7 after induction of adipocyte differentiation,but no apparent changes of β-actin,α-tubulin,peptidylprolyl isomerase A (PIPA),and 18S mRNA expressions were detected.The expression changes of key transcript factors for adipocyte differentiation such as PPARγ2,C/EBPα,and C/EBPβ were under-estimated by real time-PCR if GAPDH was chosen as the reference gene.Western blotting results showed that the GAPDH protein level increased gradually during adipocyte differentiation,especially on day 5 and 7 after adipocyte differentiation.There were no obvious changes of β-actin and α-tubulin protein expressions.(2) Berberine significantly inhibited mRNA and protein expressions of GAPDH in the process of adipocyte differentiation.GAPDH mRNA levels were reduced by 68.1％ and 66.3％ on day 5 and 7 after induction of adipocyte differentiation,but with no significant change in other reference genes.Conclusion It is not suitable for GAPDH to be used as an endogenous reference gene during 3T3-L1 adipocyte differentiation.

ObjectiveTo investigate the correlation between the level of resistin and the insulin resistance(IR) and the incorporation of microamount albuminuria(MA) of type 2 diabetes mellitus (T2DM).MethodsTwo hundred and twenty T2DM patients and 40 normal subjects (control group) were enrolled in this study.Two hundred and twenty T2DM patients were divided into 4 groups,IR group (group A,79 cases),non-IR group (group B,48 cases),IR complicating MA group (group C,51 cases),non-IR complicating MA group(group D,42 cases).The fasting serum resistin was measured by enzyme-linked immunosorbant assay (ELISA),and the fasting plasma glucose (FPG),glycosylated hemoglobin (HbA1c),fasting insulin (FINS) was also determined.ResultsThe level of fasting serum resistin in group A,B,C and D was higher than that in control group [(33.45 ± 1.37),(23.36 ± 1.47),(44.45 ± 1.39),(37.45 ±1.57) μ g/L vs.( 17.44 ± 1.26 ) μ g/L],and there was significant difference among 5 groups (P ＜ 0.01 ).The level of fasting serum resistin in group A was higher than that in group B (P ＜ 0.01 ),and the level of fasting serum resistin in group C was higher than that in group D (P ＜ 0.01 ).The level of fasting serum resistin was both higher in group C and D than that in goup A and B (P＜ 0.01 ).Correlation analysis showed the fasting serum resistin was positively correlated with body mass index(BMI),FPG,HbA1c and FINS in group A(r =0.35,0.46,0.37,0.49,P ＜0.05),and the fasting serum resistin was positively correlated with HbA1c,BMI,systolic blood pressure and diastolic blood pressure in group C(r =0.45,0.32,0.37,0.29,P ＜ 0.05 ).ConclusionsSerum resistin may participate in the process of IR and the formation of MA.It may become one of the diagnostic standard of the IR and one of the important index to estimate the MA.