Objective To study the role of phosphorylation at Serine 129 in regulating the neurotoxicity of ?-synuclein. Methods Wild type and phosphomimic mutant ?-synuclein genes were over-expressed in mouse dopaminergic cells MN9D using retrovirus. The cell viability was examined using CCK-8 assay and cell morphology was observed by immunofluorescence microscopy. Results The result of real time PCR showed that WT/?-SYN and S129D/?-SYN genes were overexpressed in MN9D as compared to uninfected MN9D and vector control group(P

Objective To establish the theoretical basis of the probiotic application among very low birth weight(VLBW) infants,the efficacy of probiotics on the gut microbiota of VLBW infants on the 14th postnatal day was studied.Method The VLBW infants admitted to BaoAn Maternal and Child Care Hospital from January 2015 to December 2015 were randomly assigned into probiotics group and placebo group.From the first feeding to corrected gestational age of 36 weeks,probiotics group was treated with a combination of Bifidobacterium and Lactobacillus while placebo group with placebo.Fecal samples were collected on the 1st and 14th postnatal day.Total bacterial DNA was extracted and sequenced using high-throughput 16S rRNA gene sequencing on MiSeq sequencing platform.Result A total of 21 VLBW infants were enrolled,9 in probiotics group and other 12 placebo group.No significant differences of clinical data existed between the two group (P ＞ 0.05),The abundance and diversity of microflora (P ＞ 0.05) on the first day between the two group were similar.Compared with placebo group,the relative abundance of Bifidobacterium and Lactobacillales in stool samples on the 14th day was significantly increased,while the Halomonas was significantly decreased.The relative abundance of the Shannon-index was increased,but without significant difference (P =0.16).Conclusion Enteral supplementation of probiotics in VLBW infants may increase probiotic bacterium and microflora diversity.

Objective To construct the recombinant rAAV2-hGAD65 vector and detect its function both in vitro and in vivo. Methods The cDNA of human glutamic acid decarboxylase 2 (hGAD65) gene, which was one of gamma-aminobutyric acid (GABA) synthetase, cloned by the method of RT-PCR, was subcloned into the adeno-associated virus (AAV) vector and formed the recombinant vector of AAV-hGAD65 (rAAV2-hGAD65). The recombinant vector was packaged by the AAV Helper-Free System and its titer was detected. The primary fibroblast, cultured from the rat lung, was transfected by the rAAV2-hGAD65. The expression of the hGAD65 in fibroblast was detected by immunohistochemical method and the level of GABA in the media was assayed by high performance liquid chromatograph (HPLC). In vivo, the hGAD65 was detected by immunohistochemical method in STN and the concentration of the GABA in the reticular part of substantia nigra (SNr) was assayed by HPLC after the rAAV2-hGAD65 was injected into the subthalamic nucleus (STN) by the stereotaxic method. Results The sequence of hGAD65 cDNA was in accordance with that in the Genebank. The amino acid sequence of hGAD65 had no mutation and the titer of rAAV2-hGAD65 was reached 4.5 ×10~(11) per milliliter. The efficiency of infection to the rat primary firoblasts was 80%, and the concentration of GABA in the media of fibroblasts was (45.66±6.07)nmol/L per liter. In vivo, hGAD65 could be detected in STN, and the concentrateion of the GABA in the SNr was increased from (5.66±1.07)nmol/g to (12.66±2.59)nmol/g.Conclusion The cDNA of hGAD65 was cloned by RT-PCR and the recombinant vector of rAAV2-hGAD65 was constructed. The AAV can infect the primary fibroblast in vitro and the hGAD65 can catalyse the glutamic acic to GABA. In vivo, the concentration of GABA in the SNr was heighten after the rAAV2-hGAD65 was injected into the STN.

Objective To assess the condition of left ventricular outflow tract obstruction (LVOTO) under resting conditions and physiological exercise in hypertrophic cardiomyopathy (HCM) patients.Methods A total of 60 patients with HCM and left ventri cular outflow tract gradient (LVOTG) ＜ 50 mm Hg (1 mm Hg =0.133 kPa) at rest were enrolled consecutively,and LVOTG at rest and exercise were measured by echocardiography.Of 51 patients with gradients ＜ 30 mm Hg at rest,26 were latent LVOTO with exercise peak value LVOTG ≥ 30 mm Hg,25 were non LVOTO with exercise peak value LVOTG ＜ 30 mm Hg,and 9 were resting obstruction with LVOTG 30-49 mm Hg.The morphological characteristics of different types of obstruction were analyzed.Results Patients with latent LVOTO were more likely to have SAM(73.1％ vs 8.0％),narrow of LVOT(46.2％ vs 4.0％),higher resting gradients [(16.9 ±7.2) mm Hg vs (7.1 ± 4.3) mm Hg] and mitral regurgitation grade at rest than patients with non-obstructive (all P values ＜ 0.05).The distribution of septal hypertrophy were different in the two groups (P ＜ 0.05).Multivariate logistic regression analysis showed independent predictors of latent LVOTO were SAM (OR 6.431,95 ％ CI 2.323-291.112,P =0.002) at rest and distribution of septal hypertrophy (OR 0.011,95％ CI 0.001-0.179,P =0.008).Conclusions Approximately half of patients with nonobstructive HCM at rest have latent LVOTO.SAM and distribution of septal hypertrophy may be useful to identify patients with latent obstruction.

Abstract: Objective To compare the screening effects of RDT, microscopy and PCR for malaria among residents in low malaria areas and elimination areas, and to investigate the presence of malaria in residents of border Villages in Cangyuan Va County and asymptomatic infections in surrounding areas, providing a basis for preventing re-introduction of malaria after elimination. Methods From August 2020 to March 2021, the fingertip blood of the investigated subjects was collected from three survey sites in the border area between China and Myanmar, namely Banlao Township in Cangyuan Va Autonomous County of Lincang City, Banwai District, Mengmao County, the Second Special Zone of Shan State, Myanmar, Yongmo and Dayan Township, Nandeng Special Zone, the Second Special Zone of Shan State, Myanmar. The malaria parasite antigen detection test kit, malaria parasite microscopic examination, fluorescent quantitative PCR and nested PCR were used to detect the asymptomatic infection of malaria parasites. Results A total of 1 040 blood samples were collected, including 606 from China and 434 from Myanmar, with 506 males and 534 females. Among them, , there were 51 individuals aged 0 to <5 years, 283 aged 5 to < years, 187 aged 15 to < years, 232 aged 30 to <45 years, 205 aged 45 to < years, and 82 aged ≥60 years. All 1 040 people tested negative for plasmodium antigen detection kit. One case of Plasmodium vivax detected by plasmodium microscopic etiology, with a detection rate of 0.10%. One case of P. vivax was also detected by fluorescent quantitative PCR and nested PCR, with a detection rate of 0.10%. Among them, one case of P. vivax was detected in Banwai District, Mengmao County, the Second Special Zone of Shan State, Myanmar, with a detection rate of 0.35%. The detection rates of malaria parasites in Banlao Township in Cangyuan Va Autonomous County of Lincang City, Yunnan Province and Yongmo Township and Dayan Township, Nandeng Special District, the Second Special Zone of Shan State, Myanmar were both 0. The difference in the detection rate of malaria parasites among the three survey sites was not statistically significant (χ2 =2.682, P>0.05). The asymptomatic P. vivax infection was detected in a 6-year-old girl from Banwai District, Mengmao County, the Second Special Zone of Shan State, Myanmar. Conclusions RDT is not suitable for malaria screening in low malaria area and elimination area. Microscopic examination and PCR can be used for malaria screening, but PCR operation is complex and costly. In surrounding areas outside of China, malaria is still prevalent, while there is no source of malaria infection in border villages of Cangyuan Va County. However, there is a risk of importation, and timely and effective measures should be taken to prevent reintroduction and transmission.

Objective:To investigate the mutation of desmosomal protein gene of arrhythmogenic right ventricular cardiomyopathy (ARVC) in people from Yunnan unexplained sudden death (YUSD) area in Xiangyun County, Dali Prefecture, Yunnan Province, and to explore the etiological relationship between the mutation of ARVC desmosomal protein gene and YUSD.Methods:The autopsy cardiac blood sample of YUSD case ( n = 1) and the peripheral venous blood samples of the same time case ( n = 1) and relatives of YUSD case ( n = 16) were collected in Xiangyun County. Blood DNA was extracted for PCR amplification and sequencing of a total of 97 exons of the ARVC desmosomal protein genes [plakophilin 2 (PKP2), junction plakoglobin (JUP), desmoplakin (DSP), desmoglein 2 (DSG2) and desmocollin 2 (DSC2)] were conducted by Sanger method. At the same time, basic information and genetic family of YUSD case, the same time case and relatives of YUSD case were investigated, and gene mutations were comprehensively analyzed. Results:The YUSD case and the same time case carried JUP, DSP and DSG2 gene mutations. Among the relatives of YUSD case, 2, 14, 16, 15 and 4 cases had mutations in PKP2, JUP, DSP, DSG2 and DSC2 genes, respectively. The YUSD case, the same time case and the relatives of YUSD case carried 6 identical mutation sites: JUP gene exon 3 c.213 T>C synonymous mutation, exon 14 c.2089 A>T missense mutation; DSP gene exon 19 c.2631 G>A synonymous mutation, exon 24 c.8472 G>C synonymous mutation; DSG2 gene exon 8 c.861 C>T synonymous mutation, and exon 15 c.3321 T>C synonymous mutation.Conclusion:In Xiangyun County, six identical mutation sites (JUP gene c.213 T>C and c.2089 A>T, DSP gene c.2631 G>A and c.8472 G>C, DSG2 gene c.861 C>T and c.3321 T>C) carried by YUSD case, the same time case and the relatives of YUSD case may be related to the incidence of some YUSD cases.

Objective To establish a LC-MS/MS method for the determination of salidroside in the capsule. Methods An electrospray ionization and multiple reaction detection were used to detect negative ion. Theophylline was used as standard. The detection ions of salidroside and theophylline used for quantitative analysis were m/z 299.0→119.0, and m/z 178.8→164.0, respectively. The Shim-pack XR-ODS (3.0 mm×75 mm, 2.0 μm) column was used for separation. The mobile phase was acetonitrile: 5 mmol/L ammonium acetate solution (90∶10, V/V). The flow rate was 0.40 ml/min. The column temperature was 25 ℃. Results The content of salidroside was analyzed within 3 minutes. The linear range was 10–2 000 ng/ml, and the minimum detection limit was 10 ng/ml. Conclusion The method has good repeatability, high sensitivity, fast analysis speed and simple operation. It can be used as a method for the determination of salidroside in the capsule. It is suitable for the quality inspection of drugs and convenient for safe use.