Background Neurotransmitter secretion disorder induced by chronic manganese poisoning has always been one of the important causes of body injury, but the mechanism of neurotransmitter secretion disorder caused by manganese is not clear at present. Objective To investigate the effects of presynaptic membrane intracellular protein 13-1 (Munc13-1) and synapse fusion protein binding protein 18-1 (Munc18-1) on dopamine secretion dysfunction induced by manganese chloride (MnCl₂) in human neuroblastoma (SH-SY5Y) cells. Methods A SH-SY5Y cell model induced by MnCl₂ was established. Cell viability was measured by MTT assay. Four experimental groups were set up: control group and low-, medium-, and high-dose manganese groups (0, 100, 200, and 400 μmol·L⁻¹ MnCl₂). They were treated with corresponding doses of MnCl₂ for 24 h. The secretion of dopamine was measured by enzyme-linked immunosorbent assay. The mRNA expression of Syntaxin-1 was detected by real-time quantitaive PCR. Total cell proteins were extracted, and the protein expression levels of Munc13-1, Munc18-1, and Syntaxin-1 were detected by Western blotting. The correlations of MnCl₂ exposure and dopamine secretion with the protein expressions of Munc13-1 and Munc18-1 were also analyzed by Pearson correlation. Results Compared with the control group, the cell viability rate decreased gradually with the increase of manganese exposure concentration, and the difference between the medium- and the high-dose manganese groups was statistically significant (P<0.05). The concentration of dopamine in cell culture medium of all manganese exposure groups decreased with the increase of manganese concentration, and compared with the control group and the low-dose manganese group, the medium- and the high-dose manganese groups were statistically significant (P<0.05). The expression of Syntaxin-1 at mRNA or protein level did not change significantly among groups (P>0.05). Compared with the control group, the protein expression of Munc13-1 decreased and that of Munc18-1 increased with the increase of manganese concentration (P<0.05). Compared with the low-dose manganese group, the changes of Munc13-1 protein in the high-dose manganese group and Munc18-1 protein in the medium- and high-dose manganese groups had statistical significance (P<0.05). Compared with the medium-dose manganese group, the protein changes of Munc18-1 in the high-dose manganese group were statistically significant (P<0.05). The correlation analysis showed that MnCl₂ dose was negatively correlated with Munc13-1 protein expression (r=−0.898, P<0.05), and positively correlated with Munc18-1 protein expression (r=0.678, P<0.05). Dopamine secretion was positively correlated with Munc13-1 protein expression (r=0.932, P<0.05), and negatively correlated with Munc18-1 protein expression (r=−0.817, P<0.05). Conclusion The inhibition of dopamine secretion in SH-SY5Y cells induced by manganese exposure is related to up-regulation of Munc18-1 and down-regulation of Munc13-1 expression levels, which may be one of the reasons for nerve injury caused by manganese.

Objective To explore the short axial strain of left ventricle in patients with atrial septal defect(ASD)before and after the transcatheter closure by using two-dimensional speckle tracking imaging (2D-STI).Methods A total of 30 patients with ASD underwent echocardiography before and after the transcatheter closure.The procedure was performed to obtain the peak of circumferential strain(Sc)and radial stain(Sr)of left ventricle by 2D-STI.Thirty healthy volunteers were enrolled as controls.Results ①Compared with the control group,regional myocardial Sc of ASD group decreased(P <0.05).Sc of anterior septum (AS),anterior wall (AW),laterior wall (LW)and posterior wall (PW)increased (P <0.05)at 2-days after the transcatheter closure and those parameters were higher than control group.At the six-months after the transcatheter closure,those parameters reduced to the normal level (P <0.05).Sc of inferior wall (IW)and mid-posterior septum (MP-Sept)increased to the normal level at 2-days after the closure surgery (P <0.05).②Compared with control group,regional myocardial Sr of ASD group decreased(P <0.05 ) except PW,two-days after the transcatheter closure these parameters increased to the normal level.Sr of PW in ASD group increased compared with control group(P <0.05),and there were no statistical changes at 2-days after the transcatheter closure.Sr of PW in ASD decreased (P <0.05 )to normal level until 6-months after the transcatheter closure.③ Global circumferential strain (GCS)of left ventricular in ASD group were lower than control group (P <0.05 ).At two-days after undergoing transcatheter closure the GCS increased (P <0.05)and those parameters were higher than control group (P <0.05).Six-months after the transcatheter closure those parameters reduced to the normal level (P < 0.05 ).There were no statistical differences of left ventricular global radial strain (GRS)between control group and ASD group. However,the GRS of ASD group increased (P < 0.05 )and those parameters were higher than control group (P <0.05)after 2-days undergoing the transcatheter closure.At six-months after the transcatheter closure those parameters were reduced to normal level (P <0.05 ).Conclusions 2D-STI can quantitative evaluate left ventricular circumferential strain and radial strain in patients with ASD before and after the transcatheter closure.

The HPLC fingerprint determination method of Jinzhen oral solution was established to provide a new method for quality control of Jinzhen oral solution. RP-HPLC was used for phenomenex Luna C18 (4.6 mm x 250 mm, 5 μm) chromatographic column, with 0.1% H3 PO4 water solution and acetonitrile as the mobile phase for gradient elution. The detection wavelength was 280 nm. HPLC fingerprint of Jinzhen oral solution was established to identify 17 common peaks in Jinzhen oral solution. The similarity of fingerprints of 10 batches of finished products was more than 0. 90. The established HPLC fingerprint has a better precision, reproducibility and stability, and can be applied in quality control of Jinzhen oral solution.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Quality Control

Objective To investigate the feasibility and clinical effectiveness of interventional ventricular septal defect(VSD) occlusion using asymmetric occluders in patients whose VSD are adjacent to aortic valves.Methods One hundred and fifty-two patients from our hospital whose VSD were occluded with asymmetric occluders were examined by echocardiography.Dimensions,shapes and shunts of VSD,structures around defects were observed and aortic valve prolapse(AVP) degrees were accessed before the interventions.The patients were followed up periodically after the procedures.The longest follow-up was 6 years.Results Sizes of VSD measured from left ventricular side were larger than those measured from right ventricular side.Twenty-seven cases with VSD had mild AVP.Right sides of defects had adhesions with variable morphological patterns in 85% patients with perimembranous VSD.After occlusion,there were 7 cases showed residual shunt,2 cases with the third degree atrio-ventricular blockages,3 cases appeared aortic-valve regurgitations (AR) and 1 case with slightly increased AR.Conclusions Immediate and medium-term effects of interventional occlusion with asymmetric occluders were satisfactory in the patients with VSD and mild AVP whose aortic valve margin were insufficient to symmetric occluders.Surgical operation should be preferred to patients with VSD and moderate to severe AVP and to very young children.Pre-closure echocardiographic evaluation is critical to the success of interventional closure with asymmetric occluder and the long-term out coming.

Objective: To investigate electronic cigarette (e-cigarette) use and its influencing factors among junior high school students of health promotion schools in Xihu District, Hangzhou City, so as to provide insights into school-based tobacco control. Methods: Grade 1 to 3 junior high school students of health promotion schools in Xihu District were recruited using a multi-stage stratified cluster sampling method from September to December, 2021. The participants' demographic features, e-cigarette use and exposure to tobacco advertising were collected using the Chinese version of the Global Youth Tobacco Survey, and the factors affecting the intention to use e-cigarettes were identified using a multivariable logistic regression model. Results: The 1 677 respondents included 875 boys (52.18%) and 802 girls (47.82%), and grade 1 to 3 junior high school students consisted of 33.93%, 35.00% and 31.07% of all respondents, respectively. There were 1 461 students that had heard of e-cigarettes (87.12%), 101 students with intention to use e-cigarettes (6.02%), and 24 current users (1.43%). Multivariable logistic regression analysis identified living in rural areas (OR=2.364, 95%CI: 1.442-3.875), having close friends that were smokers (OR=5.614, 95%CI: 3.404-9.258), having seen smoking via TV, video or movie in the past 30 days (OR=2.106, 95%CI: 1.259-3.523), having received free tobacco products (OR=3.887, 95%CI: 1.172-12.894), considering e-cigarettes as nicotine-free (OR=208.442, 95%CI: 55.713-779.856), and considering smoking making comfortable at party (OR=4.534, 95%CI: 1.853-11.090) as factors affecting intention to use e-cigarettes. There were 38.04% (638/1 677) of junior high school students with exposure to advertisements for e-cigarettes and related products, and stores, supermarkets, convenience stores, grocery, e-cigarette experience stores or offline retail stores were the primary places to contact e-cigarettes and related products. Conclusions The percentage of intention to use e-cigarettes was low among junior high school students of health promotion schools in Xihu District in 2021, and their intention to use e-cigarettes was mainly affected by close friends' smoking status and personal recognition.

Objective To establish a method for detecting HBV cccDNA in hepatocytes of chronic hepatitis B patients.Method 21 liver biopsies from the hepatic operation patients in the hospital of jiangsu province,concluding 19 HBV chronic infected patients (10 HBeAg positive patients and 9 HBeAg negative patients) and 4 uninfected patients,HBV DNA(+) serum of hepatitis B patients was thought as rcDNA.To use proteinase K to release HBV cccDNA and genomic DNA,then divide the cell lysis solution into two parts,one for detecting HBV cccDNA,the other for detecting the number of ?-Globin as internal control. Nucleic acid for detecting HBV cccDNA extracted by phenol-chloroform was digested by plasmid-safe ATP dependent DNase which was applied to digest the single strand DNA in rcDNA and ssDNA,then was quantitated by the primers spanning across the nick and SYBR Green Ⅰ dye.The specifity of PCR production was confirmed by the sequence analysis and rcDNA comparison.The significance of the difference of HBV cccDNA level between HBeAg(+) and HBeAg(-) group was analyzed by two group t test.Results The agarose gelelectrophoresis showed the molecular weight of the PCR production was about 350bp.The coincidence rate of PCR production and goal fragement was nearly 99% by sequence analysis.The result of PCR detection of rcDNA group was negative.The positive rate of HBV cccDNA of liver biopsies of HBeAg (+) patients detected by this method was 100%,the level of HBV cccDNA in the liver biopsies of HBeAg (+) patients was higher than HBeAb(+) patients.Conclusions The specificity of the method is proved by agarose electrophoresis,gene sequencing of the PCR product and rcDNA comparison.The quantitative method that use SYBR Green Ⅰ dye and ?-Globin as internal control is more specific,sensitive and economical,and more suitable for clinical purpose.

Objective To explore the method of pre expanded deltopectoral flap for repairing post burn faciocervical scars.Methods Anterior axillary incisions were made and appropriate expanders were implanted above anterior chest wall at the first stage.After a 4 6 months' expanding,the flaps based on perforating branches of the internal mammary artery,branches from the thoracoacromi al area,or perforating branches from deltoid muscle,were designed and raised according to scars and dominant vessels.The donor sites were closed at same time without skin graft.Results 43 patients with 51 flaps were operated for reconstruction of post burn faciocervical scars.All flaps and donor sites survived well.Conclusions Pre expanded deltopectoral flap is an ideal donor site for repairing post-burn faciocervical scars.

Objective To evaluate the effectiveness of prevention of dental caries in preschool children by family-based oral health management. Methods A total of 144 families with 3 years old children in Huangpu District were randomly divided equally into intervention group and control group.The intervention period was 2 years.Semi-annual oral health guidance was given to families in the intervention group(including oral examination, children′s deciduous teeth coating with fluoride, dental caries filling, oral health lectures for families, distribution of family oral health brochures, etc.)Families in the control group received annual oral examination.After the intervention, the parents completed their child′s oral health questionnaire. Results In the 2-year oral monitoring period, the incidence of dental caries in the intervention group and the control group was 9.72% and 22.22%, respectively within one year, and the difference between the two groups was statistically significant(χ²=4.19, P < 0.05).The incidence of dental caries was 12.50% and 36.11% within two years, and the difference between the two groups was also statistically significant(χ²=10.91, P < 0.01).Result of the questionnaire survey showed that children in the intervention group had better eating and oral health habits than the control group, and the differences between the two groups were statistically significant(P < 0.05). Conclusion Regular and professional family oral health management can effectively reduce the incidence of dental caries and improve the oral health in preschool children.

The aim of this study was to explore the inhibitory effect of newly synthesised emodin derivatives on the proliferation of leukemia cell lines and to select the most effective one from these emodin derivatives for further research. Emodin derivatives were synthesized by modifying the structure of emodin. MTT method was used to detect the proliferative inhibition in leukemia cell lines treated with emodin derivatives. The results showed that the half inhibitory concentration (IC50) for K562 cells treated with emodin derivatives E10-19 for 48 h was 0.84 - 12.01 µmol/L. E19 displayed the best anti-proliferative activity, while E16 and E17 did not show effects on K562 cells. Emodin derivative E19 was chosen for treating U937, NB4, Molt-4 and CA-46 cells, their IC50 for 48 h were 0.85, 0.9, 0.76, 0.8 µmol/L respectively. The IC50 of E19 for LQ2 cells was 3.60 µmol/L, and the IC50 range of E19 for normal human peripheral blood mononuclear cells at 48 h was 4.01 - 4.78 µmol/L. It is concluded that emodin derivative E19 can strongly inhibit the growth of leukemia cells and its inhibiting effect on proliferation of leukemia cells has a certain specificity. The specific mechanism of E19 anti-leukemia effect should be further studied.
Cell Proliferation ; drug effects ; Emodin ; analogs & derivatives ; pharmacology ; Humans ; K562 Cells ; Leukemia ; pathology

Objective: To explore the distribution features of microvolt T-wave alternation(MTWA) through exercise stress test (EST) in coronary artery disease (CAD) patients with MTWA changes after revascularization. Methods: MTWA was measured in pre-cordial ECG leads in 326 patients with suspected CAD. Based on coronary angiography and coronary CTA, the patients were divided into 4 groups: ① Control group, patients without coronary stenosis, n=101, ② Coronary stenosis<50％ group, n=99, ③ 50％ ≤ Coronary stenosis<70％ group, n=53 and ④ Coronary stenosis ≥ 70％ group, n=73; MTWA was compared among different groups. 95 patients with coronary stenosis ≥ 50％were further divided into 2 subgroups: R (right coronary)stenosis ≥ 50％ subgroup, n=23 and LAD (left anterior descending branch) stenosis ≥ 50％ subgroup, n=72; MTWA was respectively compared to Control group. In addition, MTWA was collected from 103 patients with percutaneous coronary intervention (PCI) as PCI group, MTWA was compared to Coronary stenosis ≥ 70％ group. Results: MTWA was obviously higher in Coronary stenosis ≥ 70％ group than the other 3 groups, all P<0.01. Compared with Control group, Rstenosis ≥ 50％ subgroup had increased MTWA in V4-V6 pre-cordial leads, P<0.05; LAD stenosis≥50％ subgroup had increased MTWA in V1-V2 pre-cordial leads, P<0.01. Compared with Coronary stenosis ≥ 70％group, PCI group showed reduced MTWA, P<0.01. Conclusion: CAD patients with severe coronary stenosis (≥70％) had increased MTWA; MTWA distribution in body surface was approximately corresponding to coronary stenosis site and PCI may decrease MTWA in CAD patients.