uate and should be standardized.

Objective To evaluate effectiveness of prevention and treatment for patients with chronic obstructive pulmonary disease (COPD) for one year at community health-care service (CHS)centers. Methods A quasi-experiment design was used to evaluate effectiveness of community management for patients with COPD who visited and registered at Xiaokunshan and Xinbang CHS centers in Songjiang district of Shanghai in 2008, with COPD patients from Xiaokunshan community CHS center as management group and those from Xinbang as control. Measures for community management included training for local general practitioners in essential knowledge of COPD prevention and treatment, allocation of necessary drugs for COPD treatment, and health education for COPD patients and their family members. No special measures were taken for control group except routine treatment. Results A total of 132 patients were diagnosed as COPD according to their pulmonary function on 2008, 61 at Xiaokunshan and 71 at Xinbang CHS canters.One hundred and two patients, 47 at Xiaokunshan and 55 at Xinbang, finished one-year follow-up. Five patients from Xiaokunshan who were smokers at their first visits and three of them quitted smoking after oneyear management. Meanwhile, none of nine smoked patients from Xinbang quitted smoking in the same period. There was no statistically significant difference in symptom scores, pulmonary function, and forced expiratory volume at the first second (FEV1) between patients in the two groups during one-year follow-up.Average score of quality of life in patients at Xiaokunshan dropped to 39. 12 after one-year management from 46. 96 at their first visits, and that in patients at Xinbang increased to 62. 11 from 56. 55 (P＜0.01).Average six-minute walking distance (6-MWD) in patients at Xiaokunshan reduced to 354. 26 meters after one-year management from 361.66 meters at their first visits, meanwhile that in patients at Xinbang reduced to 351.18 meters from 398.07 meters (P =0. 008). Scores of functional dyspnea in patients at Xiaokunshan fell to 0.34 from 0.40 at their first visit, meanwhile that in patients at Xinbang increased to 1.00 from 0.95(P =0.038). During the one-year follow-up, 13 patients at Xiaokunshan and 53 at Xinbang visited emergency departments for treatment, and four at Xiaokunshan were hospitalized and none died, and five at Xinbang were hospitalized due to acute exacerbation of COPD and four of them died. Conclusions The study suggests that strengthening prevention and treatment for COPD patients at community level by general practitioners, patients themselves and their family members can improve their quality of life, reduce emergency visits and hospitalization, alleviate dyspnea symptoms and delay the decline of 6MWD.

In this paper,we have made a preliminary analysis on the necessity,main contents and difficulties in teaching medical undergraduates the principles and applications of optogenetics experimental technology.We focused on theory and practice teaching and emphasized on the most important technological principles,including how to achieve selective intervention of specific neural pathways;how to achieve twoway control of cell excitation or inhibition effect and the main process of the experiment.Finally,we have summarized our recent experience in the teaching process,to provide reference for basic medical experiments teaching in related medical colleges,so as to promote optogenetics experimental teaching for undergraduates in domestic medical colleges.

Objective:To evaluate the effect of orexin-A on programmed necrosis during cerebral ischemia-reperfusion (I/R) in rats.Methods:Thirty clean-grade healthy adult male Spraugue-Dawley rats, weighing 280-320 g, were divided into 3 groups ( n=10 each) using a random number table method: sham operation group (Sham group), cerebral I/R group (I/R group) and orexin-A group (OA group). In I/R and OA groups, a rat model of global cerebral I/R injury was established by transesophageal cardiac pacing-induced cardiac arrest and cardiopulmonary resuscitation in anesthetized animals.Orexin-A 30 μg/kg (diluted to 0.5 ml in phosphate buffer solution) was intravenously injected at 10 min before establishing the model in OA group.Phosphate buffer solution 0.5 ml was intravenously injected at 10 min before establishing the model in Sham and I/R groups.The neurological deficit score (NDS) was assessed at 24 h of reperfusion, then the rats were sacrificed, and bilateral hippocampal tissues were obtained.The morphological structure of pyramidal cells in the hippocampal CA1 region was examined after HE staining, and normal pyramidal cells were counted.Western blot was used to detect the expression of receptor-interacting protein 1 (RIP1), RIP3 and mixed-lineage kinase domain-like protein (MLKL). Immuno-histochemistry was used to count RIP1, RIP3 and MLKL positive cells in the hippocampal CA1 region.The activity of superoxide dismutase (SOD) in hippocampi was determined by xanthine oxidase method, and the content of malondialdehyde (MDA) in hippocampi was determined by thiobarbituric acid method. Results:Compared with Sham group, the normal pyramidal cell count in the hippocampal CA1 region was significantly decreased, NDS was increased, the expression of RIP1, RIP3 and MLKL protein was up-regulated, the positive cell count was increased, the content of MDA in hippocampi was increased, and the activity of SOD in hippocampi was decreased in I/R and OA groups ( P<0.05). Compared with I/R group, the count of normal pyramidal cells in the hippocampal CA1 region was significantly increased, NDS was decreased, the expression of RIP1, RIP3 and MLKL protein was down-regulated, the count of positive cells was decreased, the content of hippocampal MDA was decreased, and the activity of hippocampal SOD was increased in OA group ( P<0.05). Conclusion:The mechanism by which orexin-A reduces cerebral I/R injury may be related to inhibiting programmed necrosis in rats.

Objective To investigate the effect of Kinesio Taping on interleukin-1β(IL-1β),tumor necrosis factor α(TNF-α)and matrix metalloproteinase-3 (MMP-3)in synovial fluid of patients with knee osteoarthritis in early and middle stage.Methods A total of 84 patients with knee osteoarthritis who met the inclusion/exclusion criteria were randomly selected and divided into the control group treated with placebo(kinesiology tape without elasticity)and the experimental group treated with Kinesio Tape.Both two tapes were changed every two days,with 15 times of changes as a course of treatment.Synovial fluid sample was drawn before and one week after treatment,and was used for measuring Levels of 1L-1β,TNF-α and MMP-3 by enzyme-linked immunosorbent assay (ELISA).and knee function was evaluated before and 1 week after treatment.Results Compared with pre-treatment,the levels of IL-1β,TNF-α and MMP-3 in synovial fluid were significantly decreased in the experimental group after treatment [(20.07 ± 6.94) ng/Lvs.(38.12 ± 5.93) ng/L,(42.42±8.76)ng/Lvs.(58.23±9.54)ng/L,(11.28±1.99)μg/L vs.(15.67±2.21)μg/L,t =12.81,7.91 and 9.57,all P＜0.05].The levels of IL-1β,TNF-α and MMP-3 in synovial fluid were lower in the experimental group after treatment than before treatment[(20.07±6.94)ng/L vs.(38.12±5.93)ng/L,(42.42±8.76)ng/L vs.(58.23±9.54)ng/L,(11.28 ±1.99)μg/L vs.(15.67±2.21)ng/L,t =12.81,7.91 and 9.57,all P＜0.05],and were lower in the experimental group after treatment than in control after treatment[(20.07±6.94)ng/L vs.(37.97±6.21)ng/L,(42.42±8.76)ng/L vs.(57.04 ±8.73)ng/L,(11.28± 1.99)μg/Lvs.(15.01± 2.56)μg/L,t =12.46,7.66 and 7.46,all P＜0.05]Lysholm score was significantly higher in the experimental group after treatment than before treatment[(74.5 ± 2.6) vs.(44.7 ± 2.8),t =50.54,P ＜0.05],and was also higher in the experimental group after treatment than in the control group after treatment[(74.5±2.6) vs.(50.2± 2.3),t =45.37,P＜0.05].Conclusions Kinesio Taping can significantly reduce the levels of IL-1β,TNF-α and MMP-3 in synovial fluid of patients with knee osteoarthritis through inhibiting the inflammatory response in articular cavity.

OBJECTIVETo investigate the effect of poly-ADP-ribosylation in hexavalent chromium Cr(VI) induced cell damage.

METHODSThe study object, poly (ADP-ribose) glycohydrolase (PARG) deficient human bronchial epithelial cells (16HBE cells), was constructed previously by our research group. Normal 16HBE cells and PARG-deficient cells were treated with different doses of Cr (VI) for 24 h to compare the differences to Cr (VI) toxicity, meanwhile set up the solvent control group. On this basis, 5.0 µmol/L of Cr (VI) was selected as the exposure dose, after the exposure treatment, total proteins of both cells were extracted for two dimension fluorescence difference gel electrophoresis (2D-DIGE) separation, statistically significant differential protein spots were screened and identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS/MS), and further validated by Western blot.

RESULTSAfter Cr (VI) treatment, the survival rate of PARG-deficient cells was higher than normal 16HBE cells. When the doses reached up to 5.0 µmol/L, the survival rate of 16HBE cells and PARG-deficient cells were respectively (59.67 ± 6.43)% and (82.00 ± 6.25)%, the difference between which was significant (t = -4.32, P < 0.05). 18 protein spots were selected and successfully identified after 2D-DIGE comparison of differential proteins between normal 16HBE cells and PARG-deficient cells before and after exposure. The function of those proteins was involved in the maintenance of cell shape, energy metabolism, DNA damage repair and regulation of gene expression. The differential expression of cofilin-1 was successfully validated by Western blot. The expression level of cofilin-1 in the 16HBE cells increased after Cr (VI) exposure with the relative expression quantity of 1.41 ± 0.04 in treated group and 1.00 ± 0.01 in control group, the difference of which was statistically significant (t = -18.00, P < 0.05), while the expression level in PARG-deficient cells had no statistically significant difference (t = -8.61, P > 0.05).

CONCLUSIONMost of the identified differential proteins are closely related to tumorigenesis, suggesting that poly-ADP-ribosylation reaction may resist the cytotoxicity of Cr(VI) by inhibiting Cr (VI) induced tumorigenesis, which provides important reference data to clarify the mechanisms of poly-ADP-ribosylation in Cr (VI) induced cell damage.

Bronchi ; Cell Transformation, Neoplastic ; genetics ; Chromium ; Cofilin 1 ; DNA Repair ; Epithelial Cells ; Glycoside Hydrolases ; deficiency ; physiology ; Humans ; Tandem Mass Spectrometry

OBJECTIVETo reveal the role of poly-ADP-ribosylation and DNA methylation in carcinogenic process induced induced by Cr (VI), and to discuss the relations between them.

METHODSThe pre-established Poly (ADP-ribose) glycohydrolase (PARG) deficient cells and 16HBE cells were treated with different concentrations of Cr (VI), and the changes of total genomic DNA methylation level in different groups were detected by methylation immunofluorescent detection, as well as the changes of the activity of methyltransferases. Moreover, RT-PCR and western blotting method were applied to analyze the changes of expression of DNMT1, DNMT3a, DNMT3b and MBD2, upon the protein level.

RESULTSAfter treated by Cr(VI) for 24 h, the healthy 16HBE cells showed a significant lower level of genomic DNA methylation; however, there was no significant changes (P > 0.05) found in PARG deficient cells by immunofluorescence assay. When the dose of Cr (VI) reached 5.0 µmol/L, the activity of methyltransferases in 16HBE cells and PARG deficient cells (49.33 ± 2.65, 80.05 ± 2.05) decreased by 20% and 50% comparing with contrast group (99.27 ± 1.10, 99.30 ± 0.60) . After treated by Cr (VI) for 24 h, the expression of mRNA and protein level among DNMT1, DNMT3a, DNMT3b and MBD2 decreased significantly in healthy 16HBE cells; and the expression of DNMT1 and DNMT3a decreased in PARG deficiency cells. The relevant expression levels of mRNA of DNMT1 were separately (0.99 ± 0.09), (0.79 ± 0.10), (0.59 ± 0.13) and (0.39 ± 0.02) (F = 247.17, P < 0.01), the expression levels of protein were separately (1.00 ± 0.03), (0.69 ± 0.15), (0.65 ± 0.10) and (0.55 ± 0.13) (F = 214.12, P < 0.01), the expression levels of DNMT3a mRNA were separately (1.00 ± 0.04) , (0.93 ± 0.11) , (0.79 ± 0.07) , (0.59 ± 0.05) (F = 498.16, P < 0.01) , and the expression levels of protein were separately (1.00 ± 0.14) , (0.97 ± 0.11) , (0.79 ± 0.17) , (0.57 ± 0.15) (F = 390.11, P < 0.01) when the dose of Cr (VI) at 0, 0.3, 1.2 and 5.0 µmol/L. However, there were no significant changes of expression found in DNMT3b and MBD2.

CONCLUSIONPoly-ADP-ribosylation could regulate the activity of DNMT3b and MBD2, protect cells against the DNA methylation alteration induced by Cr(VI) and maintain the global genomic DNA methylation level.

Cell Line ; Chromium ; toxicity ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; metabolism ; DNA Methylation ; drug effects ; DNA-Binding Proteins ; metabolism ; Epithelial Cells ; metabolism ; Genome ; Humans ; Poly Adenosine Diphosphate Ribose ; metabolism ; RNA, Messenger ; genetics

OBJECTIVETo investigate the levels and influencing factors of phthalate internal exposure in pregnant women (gestation age ≤ 16 weeks).

METHODSDuring April to June in 2013, 1 020 pregnant women (gestation age ≤ 16 weeks) who had established the maternal care manual were recruited in maternal and child health hospital of Siming District, Xiamen city. Participators were asked to complete a questionnaire to obtain information on socio-demographic characteristics, lifestyle behaviors, and antenatal examination and to provide a urine sample. Finally, 998 pregnant women who provided a urine sample and completed the questionnaire were enrolled. Adopting systematic sampling method, 100 ones were selected randomly among 998 pregnant women. High performance liquid chromatography-electrospray ionization-tandern mass was used to determine the concentration of five phthalate monoesters in each urine, including mono-n-methyl phthalate (MMP), mono-ethyl phthalate (MEP), mono-butyl phthalate (MBP), mono-benzyl phthalate (MBzP), mono-ethylhexyl phthalate (MEHP). Based on the measurements and questionnaire data, multivariate logistic regression was used to analyze the association between the phthalate monoester levels and potential influential factors.

RESULTSThe detection rates of MMP, MEP, MBP, MBzP and MEHP in 100 pregnant urine samples were 94%, 93%, 87%, 83%, 99%, respectively. And the urinary median uncorrected concentrations of MMP, MEP, MBP, MBzP and MEHP in 100 urine samples were 20.56, 17.62, 10.15, 2.03, and 5.12 ng/ml, respectively. Specific gravity-corrected concentration were 20.81, 20.36, 12.88, 2.58, 5.00 ng/ml, respectively. The results of multivariate logistic regression analysis indicated that: education degree was negatively associated with urinary concentration of MMP, MEP, MBP, MBzP and MEHP, OR (95% CI) were 0.495 (0.253-0.966), 0.380 (0.191-0.755), 0.379 (0.186-0.774), 0.401 (0.196-0.819), 0.373(0.183-0.762), respectively. Participants who had hair permed and dyed during pregnancy had higher urinary level of MBP and MBzP, OR (95% CI) were 12.867 (1.240-133.525), 15.982 (1.367-186.911), respectively; Participants who use cosmetics during pregnancy had higher urinary level of MEP and MBP, OR (95% CI) were 2.977 (1.012-8.757), 4.440 (1.485-13.272), respectively; plastic bottled water consumption was positively associated with urinary concentrations of MEP and MEHP, OR (95% CI) were 3.780 (1.417-10.083), 2.699 (1.039-7.010), respectively; annual household income was negatively associated with urinary concentration of MMP, OR (95% CI) was 0.597 (0.372-0.959); individuals who took medications during pregnancy had higher urinary level of MEHP than non-takers, OR (95% CI) was 4.853 (1.084-21.732).

CONCLUSIONPregnant women whose gestation age was less than 16 weeks are generally exposed to phthalate. Phthalate internal exposure levels are significantly associated with most measured factors and the influencing factors with different phthalates internal exposure levels are different.

Chromatography, High Pressure Liquid ; Dibutyl Phthalate ; urine ; Female ; Humans ; Life Style ; Maternal Exposure ; Phthalic Acids ; urine ; Pregnancy ; Surveys and Questionnaires ; Tandem Mass Spectrometry

Objective To analyze the clinical courses and outcomes of COVID-19 cases and the influencing factors in Guangdong province and provide basis for the formulation or adjustment of medical care and epidemic control strategy for COVID-19. Methods We collected demographic data, medical histories, clinical courses and outcomes of 1 350 COVID-19 patients reported in Guangdong as of 4 March 2020 via epidemiological investigation and process tracking. Disease severity and clinical course characteristics of the patients and influencing factors of severe illness were analyzed in our study. Results Among 1 350 cases of COVID-19 cases in Guangdong, 72 (5.3%) and 1049 (77.7%) were mild and ordinary cases, 164 (12.1%) were severe cases, 58 (4.3%) were critical cases and 7 (0.5%) were fatal. The median duration of illness were 23 days ( P 25 - P 75 : 18-31 days) and the median length of hospitalization were 20 days ( P 25 - P 75 : 15-27 days). For severe cases, the median time of showing severe manifestations was on the 12th day after onset ( P 25 - P 75 : 9th to 15th days), and the median time of severe manifestation lasted for 8 days P 25 - P 75 : 4-14 days). Among 1 066 discharged/fetal cases, 36.4% (36/99) and 1.0% (1/99) of the mild cases developed to ordinary cases and severe cases respectively after admission; and 5.2% (50/968) and 0.6% (6/968) of the ordinary cases developed to severe cases, and critical cases respectively after admission. In severe cases, 11.4% developed to critical cases (10/88). The influencing factors for severe illness or worse included male (a HR =1.87, 95% CI : 1.43-2.46), older age (a HR =1.67, 95% CI : 1.51-1.85), seeking medical care on day 2-3 after onset (a HR =1.73, 95% CI : 1.20-2.50) pre-existing diabetes (a HR =1.75, 95% CI : 1.12-2.73) and hypertension (a HR =1.49, 95% CI : 1.06-2.09). Conclusions The course of illness and length of hospitalization of COVID-19 cases were generally long and associated with severity of disease clinical outcomes. The severe cases were mainly occurred in populations at high risk. In the epidemic period, classified management of COVID-19 cases should be promoted according to needs for control and prevention of isolation and treatment for the purpose of rational allocation of medical resources.

Objective: To provide effective reference of laboratory detection and prevention-control in avian influenza epidemic via analyzing the detection result of the first case infected avian influenza H5N6 virus in Fujian province. Methods: The viral RNA was extracted from the patient’s throat swab and specimens of surrounding environment, and detected by real-time RT-PCR. The gene sequences of HA and NA gene segments were obtained by RT-PCR and sequencing, the evolution characteristics of the virus were elementarily analyzed by bioinformatics. Results: The avian influenza H5N6 virus was confirmed from the patient’s throat swab, termed influenza A/Fujian-Sanyuan/21099/2017(H5N6)virus. The throat swabs of case from 5 different time points were collected and the H5N6 nucleic acid were detected from the first three times collection. Among 43 specimens of surrounding environment, there were 16 H5 virus samples. The HA and NA gene segments of A/Fujian-Sanyuan/21099/2017 were closely related to A/Cygnus atratus/Hubei/2Z2-O/2016(H5N8) and A/chicken/Hubei/ZYSJF16/2016(H5N6), with a similarity of 99.6% and 99.0% respectively. The cleavage site of HA gene contained multiple basic amino acids. Conclusions The suspected case was the first case infected with avian influenza H5N6 virus in Fujian province, and the HA and NA genes of virus were highly similar to those of H5N8 and H5N6 virus respectively.