Objective:To detect gene mutations in a pedigree with dominant dystrophic epidermolysis bullosa (DDEB) .Methods:A 20-year-old male proband presented with repeated blisters, ulceration, pigmentation, scars on the limbs, and deformation of the nails/toenails after birth. There were 5 patients in the 3-generation family, and they all presented with typical skin lesions. Peripheral blood samples were obtained from 14 members of the pedigree (including the 5 patients) and 100 unrelated healthy controls. Whole-exome sequencing was performed in the proband to identify relevant mutation sites, which were then confirmed in the family by Sanger sequencing.Results:Genetic testing indicated that the proband and the other 4 patients all carried a missense mutation (c.7885G>A) in exon 107 of the COL7A1 gene, resulting in the substitution of glycine by arginine at amino acid position 2629 (p.G2629R). The mutation was identified neither in the 9 healthy relatives nor in the 100 unrelated healthy controls. The mutation co-segregated with DDEB in the family, and was not included in databases such as Pubmed, HGMD or ClinVar, suggesting it was a novel missense mutation. The amino acid encoded by this mutation may alter the structure of type Ⅶ collagen, thereby affecting its function.Conclusion:A novel missense mutation was identified in exon 107 of the COL7A1 gene in the family with DDEB, expanding the spectrum of mutations in the COL7A1 gene.

Background At present, occupational poisoning has become one of the most serious occupational diseases that jeopardize the health of workers in China, in addition to pneumoconiosis, with a wide range of impacts and heavy social and economic losses. Objective To analyze the characteristics and patterns of the incidence of occupational poisoning in Tianjin from 2006 to 2020, and to provide a scientific basis for the development of effective intervention measures and prevention strategies. Methods The Occupational Diseases and Health Hazards Monitoring Information System, a module of the China Disease Prevention and Control Information System, was the primary data source of the study. The case files of acute and chronic occupational poisoning diagnosed by occupational disease diagnostic institutions in Tianjin from January 1, 2006 to December 31, 2020 were retrieved from the system. The data included basic information, occupational history, disease characteristics, and employer information. Excel 2016 was used to establish database, and frequencies and composition ratios were reported. Results From 2006 to 2020, a total of 234 cases of occupational chemical poisoning were reported in Tianjin, including 49 cases of acute occupational poisoning (20.94%) and 185 cases of chronic occupational poisoning (79.06%). The number of acute occupational poisoning cases showed a rising and then falling trend, and the number of chronic occupational poisoning cases showed an overall decreasing trend. Acute occupational poisoning mainly occurred in the four districts around the city (22 cases, 44.90%), and chronic occupational poisoning mainly occurred in Binhai New Area (90 cases, 48.65%). More male cases were reported than female cases. The age of onset was concentrated at 45-55 years old. The acute occupational poisoning cases were concentrated in the group with less than 5 years of service (22 cases, 44.90%), mainly asphyxiating gas poisoning, and the highest incidence was reported in chemical raw materials and chemical products manufacturing industry. The chronic occupational poisoning cases were concentrated in the groups with a working age of 5-14 years (70 cases, 37.84%), mainly organic solvent poisoning, and the highest incidence was also reported in chemical raw materials and chemical products manufacturing industry. Conclusion The prevention and control of occupational poisoning as well as health surveillance and management in Tianjin shall be carried out in a hierarchical and focused manner in accordance with types of acute and chronic occupational poisoning chemicals in the city, regional economic differences, types of industries, and distribution of workers.

Sports fatigue of the lower limbs is one of the important factors affecting sports performance. How to improve the anti-fatigue ability of the lower limbs during endurance exercise is the focus of the research field of human sports biomechanics. This study systematically reviewed the relevant literature on transcranial direct current stimulation (tDCS) intervention on lower limb endurance performance, summarized the effect of tDCS on lower limb endurance performance, and analyzed the influencing factors and potential mechanisms. The results showed that: tDCS intervention has a significant effect on the endurance performance of the whole lower limbs, but there is no unified conclusion on the effect on the endurance performance of the knee joint. The researchers deem that tDCS can increase the excitability of the primary motor cortex and reduce the activation of the supplementary motor area and the premotor area to producing a lower rating of perceived exertion, but cannot affect the perception of exercise-induced pain, and stimulation protocols varied across studies, which may be partly responsible. This study can provide a theoretical basis for exploring the central mechanism of tDCS to improve endurance performance, formulating rehabilitation and sports training programsfor different groups of people, and developing new stimulation equipment to enhance the human body’s anti fatigue ability.

Objective To study the effects and mechanisms of molecular targeted drug combination on multi-driven proliferation hepatocellular carcinoma SK-Hep-1 cells. Methods Four molecular targeted drugs (HG6-64-1, Dasatinib, Crizotinib, and Sunitinib) were used to treat SK-Hep-1 cells, and the monophasic kinetic analysis curve and two-phase analysis curve were drawn. Western blot analysis was used to detect the effects of the above drugs on key signaling pathways in SK-Hep-1 cells. MTT assay was used to detect the effects of the above drugs and their combination on the proliferation of SK-Hep-1 cells. Results Compared with the monophasic kinetic analysis curve, the biphase analysis curve could better fit the effects of molecular targeted drugs on SK-Hep-1 cells, which predicted that the combination of HG6-64-1, Dasatinib, and MK-2206 could effectively inhibit the proliferation of SK-Hep-1 cells. Conclusion Two-phase kinetic analysis can quantitatively describe the response of multi-driven proliferation hepatocellular carcinoma SK-Hep-1 cells to molecular targeted therapy. The combination of HG6-64-1, Dasatinib, and MK-2206 is a potential drug combination for the treatment of hepatocellular carcinoma.

HYD-PEP06, an endostatin-modified polypeptide, has been shown to produce effective anti-colorectal carcinoma effects through inhibiting epithelial-mesenchymal transition (EMT). However, whether HYD-PEP06 has similar suppressive effect on hepatocellular carcinoma (HCC) remained unknown. In this study, HYD-PEP06 inhibited metastasis and EMT but not proliferation

Objective To understand the status of fluoride content in brick tea (pressed tea) in China,and conduct health risk assessment according to the situation of drinking brick tea in fluorosis areas.Methods Twenty national designated brick tea manufacturers were selected to collect 1-5 varieties of brick tea samples of mainstream varieties with large production volume and large sales volume.If low-fluoride brick tea was produced,an additional sample would be collected to determine the fluoride content of tea.The retrospective analysis method was used to collect the monitoring data of brick tea-type fluorosis in the whole country in 2017,and analyze the use of brick tea in the residents of the brick tea-type fluorosis areas in six provinces (regions) of Inner Mongolia,Sichuan,Gansu,Ningxia,Xinjiang and Qinghai.According to the fluoride content of brick tea,combined with daily average consumption data of brick tea in the brick tea-type fluorosis areas,daily average brick tea intake,and hazard quotient of the daily average brick tea fluoride and the standard recommended fluoride intake were calculated.A health risk assessment was conducted based on the hazard quotient (hazard quotient ＞ 1 indicated a health risk).Through the total fluoride intake standard of the Chinese population,the safe consumption of brick tea with different fluoride content was simulated by using the value of the hazard quotient.The tea fluoride was measured by an ion selective electrode method.Results A total of 54 samples were collected,of which 49 were brick tea samples and 5 were low-fluoride brick tea samples.The geometric mean of the fluoride content of 49 brick tea samples was 460 mg/kg,the median was 430 mg/kg,and the range was 102-1 797 mg/kg.There were 14 samples with a fluoride content ＜ 300 mg/kg,accounting for 28.6％.The average fluoride content of 5 low-fluoride brick teasampleswas 193 mg/kg,ranging from 102 to 290 mg/kg.Forty-nine brick tea samples were distributed in 5 provinces,the fluoride content of brick tea was the lowest in Yunnan,128 mg/kg;the highest in Hubei,712 mg/kg.The lowest fluoride content of brick tea in different varieties was Tuo tea,130 mg/kg,and the highest was Jinjian tea,745 mg/kg.There were 1 000 households drinking brick tea in the brick tea-type fluorosis areas in 6 provinces,73.9％ (739/1 000) of households drinking the products of the national assigned brick tea manufacturers;a total of 1 000 brick tea samples were collected,and the geometric mean of brick tea fluoride content was 551 mg/kg,and only 17.5％ (175/1 000) of brick tea had a fluoride content ＜ 300 mg/kg.Compared with the national standards and the standards of World Health Organization,hazard quotient of agriculture,animal husbandry,semi-pastoral industry,urban residents through the brick tea fluoride was 0.34,1.38,0.75,and 0.21,respectively;0.29,1.21,0.66,and 0.19,respectively.Animal husbandry hada health risk through drinking brick tea among them.According to the health standard of total fluoride intake in China,when drinking brick tea with fluoride contents of 300,430,460 and 500 mg/kg,the safe consumption of brick tea for school-age children was 2.9,2.0,1.9 and 1.8 kg,respectively,and for adult was 4.3,3.0,2.8 and 2.6 kg,respectively.Conclusions Brick tea has high fluoride content,and percent of qualified brick tea is low and few of low-fluoride brick tea are consumed.There is a health risk in the animal husbandry who has a large amount of brick tea.

Objective To analyze the voluntary blood donors of Shaoguan city in recent 10 years HIV infection status and distri‐bution characteristics ,and to take effective prevention and control measures ,in order to reduce the risk of HIV infection through blood transfusion .Methods HIV screening test in the early by the reagent of two different manufacturers ,from 2005 to 2014 vol‐untary blood donors specimens of 265 771 doses of blood donation in Shaoguan ,and early screening positive specimens confirmed by Center for Disease Control of Shaoguan .Results The voluntary blood donors of Shaoguan in the 10 years that initial screening test anti‐HIV positive were 310 cases ,and 35 cases confirmed positive;The confirmed positive rate was 1 .317/10 000;The HIV infec‐tion rate in 2012 was highest (2 .867/10 000) and there were 9 cases ,which was significant difference (P<0 .05) .The district of the maximum rate was New Fengxian ,which was 5 .369/10 000 ,and different districts had significant difference (P<0 .05) .The epidemiology survey about HIV infections found that male were 30 cases(85 .71% ) ,which was significantly more than women;By age distribution ,most HIV infections were 18 to 35 years old young adults ,which accounted 26 cases(74 .29% ) .Married and un‐married people infected respectively accounted 28 .57% and 71 .43% .The occupations of HIV infections were scattered with farm‐ers ,staffers ,the service industry ,cadres in the majority .The HIV infection rate with low education was relative higher ,which had some relationship to educational level .The residents and the non‐residents infected respectively accouted 71 .43% and 28 .57% ;Blood donors for the first time of HIV infections were 24 cases (68 .57% ) .Blood donors for the second time of HIV infections were 11 cases (31 .43% );Other virus co‐infection respectively were 2 HBV infections ,1 HCV infection and 3 syphilis infections .The route of transmission was mainly through sexual transmission that was mainly heterosexual transmission ,which was 33 cases (94 .58% ) .Conclusion The voluntary blood donors of Shaoguan with HIV is on the rise ,and give priority to with male youths spread through heterosexual sex .It should be pay high attention to by the government ,society and blood stations ,and strengthen the propaganda work of AIDS prevention and control checks ,and minimize the risk of HIV transmission by blood transfusion in or‐der to guarantee the safety of clinical blood transfusion .

BACKGROUNDHerpes simplex virus thymidine kinase phosphorylates ganciclovir to ganciclovir monophosphate, which is then converted to ganciclovir triphosphate by endogenous cellular nucleoside kinases. The ganciclovir triphosphate acts as a DNA chain terminator due to the lack of a functional 3'-OH group and terminates the process of DNA replication, hence leading to cell apoptosis. At present, HSVtk gene usually acts as suicide gene to kill tumor cells. The aim of this study was to investigate the selective cytotoxicity of the herpes simplex virus thymidine kinase/ganciclovir (HSVtK/GCV) suicide gene system controlled by the a-fetoprotein (AFP) promoter on hepatocellular carcinoma (HCC) cells in vitro.

METHODSpAFP-HSVtk-IRES2-EGFP recombinant plasmid vectors driven by the AFP promoter were constructed. HL-7702 liver cells, HUH-7 HCC, and HepG2 HCC were transfected with the recombinant plasmids. HSVtK gene expression was detected using Western blotting analysis. HepG2 cells line stably expressing HSVtk gene was selected by G418 reagent. The cytotoxicity of HSVtK/GCV suicide gene system on hepatoma cells was measured by CCK-8 reagents when different doses of ganciclovir were added.

RESULTSPlasmid pAFP-TK-IRES2-EGFP-expressed HSVtk gene was constructed successfully. HSVtk gene expression level was significantly higher in AFP-positive hepatoma cells than in AFP-negative liver cells. After G418 selection, a HepG2 cells line stably expressing HSVtk gene was acquired. With the increase of the dose of ganciclovir the optical density at 450 nm of HepG2 cells stably expressing HSVtk gene gradually decreased (P < 0.05).

CONCLUSIONThe HSVtK gene-specific expression in hepatoma cells as well as the cytotoxicity of the suicide gene system in HepG2 cells provided the basis for the targeted gene therapy of HCC.

Apoptosis ; drug effects ; genetics ; Carcinoma, Hepatocellular ; genetics ; Cell Line, Tumor ; Fetal Proteins ; genetics ; Ganciclovir ; analogs & derivatives ; pharmacology ; Hep G2 Cells ; Humans ; Liver Neoplasms ; genetics ; Promoter Regions, Genetic ; genetics ; Transfection

BACKGROUND:A group of nuclease-like proteins were previously purified from Eisenia foetida tissues, exploring primary structures of these proteins wil help to uncover basic structure characteristics of them and provide foundations for the study addressing the relationship of their structures and functions.
OBJECTIVE:To explore primary structures of nuclease-like proteins EWD1 and EWD2.
METHODS:Edman degradation method was used to sequence the N-terminal amino acids of EWD1 and EWD2, acid hydrolisis method was used to analyze amino acid compositions of EWD1 and EWD2, LC-MS/MS was used to analyze some peptide sequences within the proteins, and MALDI-TOF-MS was used to calculate the number of the disulfide bonds and the contents of polysaccharides.
RESULTS AND CONCLUSION:Among the amino acid compositions in EWD1 and EWD2, the sum contents of aspartate and asparagines were the highest (al nearly 10%), the contents of hydrophobic amino acids were also high, and the contents of cysteine was low. The EWD1 and EWD2 had similar amino acid compositions with other nucleases. Edman degradation results showed that, the N-terminal sequences of the large subunit of EWD1 were in turn as fol ows:D, E, W, V, Y, P;the N-terminal sequences of EWD2 were as fol ows:L, L, G, P, Y, K, P, K, C. The results of LC-MS/MS indicated the two proteins were novel proteins;MALDI-TOF-MS results showed that 8 cysteine residues formed 4 disulfide bonds in EWD1, 6 cysteine residues formed 3 disulfide bonds in EWD2. EWD1 and EWD2 were al glycoproteins, the content of polysaccharides was 17.3%in EWD1 and 15.6%in EWD2.

Objective To investigate the effects of Triptolide on apoptosis of cultured rat mesangial cells treated by TGF-β1 and the role of Smac in this process. Methods The mesangial cells were pre-treated with different concentrations of Triptolide for 24 hours, then stimulated with TGF-β1 for 24 hours. Apoptotic cells were detected by TUNEL assay. Smac transcription level was determined by Real time-PCR analyses. Smac expression level was assessed using Western blot anal-yses. Localization of Smac was shown by confocal fluorescence microscopy. Results Compared with control group, TGF-β1 inhibited apoptosis and Smac transcription and expression in rat mesangial cells. By contrast, Triptolide promoted mesangial cells apoptosis. In Triptolide groups, Smac mRNA and protein levels were up-regulated. Additionally, in normal and TGF-β1 groups Smac protein was mainly localized in mitochondriawhile in Triptolide groupit was mainly localized in cytoplasm and nucleus with increased fluorescence intensity. Conclusion Triptolide could promote the effect that TGF-β1 inhibited apop-tosis of mesangial cells, through both up-regulation the expression of Smac and stimulating it translocation from mitochon-dria to cytoplasm and nucleus.