Gastroesophageal reflux disease （GERD） is a frequently and commonly occurring disease in clinic. In recent decades， with the development in pathophysiology and drug researches， modern medicine has achieved remarkable progress and results in diagnosis and treatment. However， the treatments for non-erosive reflux disease， refractory gastroesophageal reflux disease， proton pump inhibitor resistance， overlap of disease symptoms， and extraesophageal symptoms are limited and ineffective. Traditional Chinese medicine （TCM） was widely used in clinical practice， which has been proved effective in relieving symptoms and improving the quality of life. Sponsored by China Association of Chinese Medicine （CACM） and undertaken by the Spleen and Stomach Disease Branch of CACM， "the 12^th Youth Salon of Clinical Predominance Disease Series （GERD）" invited 18 authoritative digestive experts of TCM and western medicine to discuss "the difficulties of clinical diagnosis and treatment of GERD and TCM advantages". The focus issues such as modern medical diagnosis and treatment achievements and contributions， improvement and maintenance of symptoms， response to overlapping disease symptoms， reduction and withdrawal of acid suppressors， and treatment of extra-esophageal symptoms were discussed in depth. TCM and western medicine exchanged and complemented each other's strengths， combing the difficulties of modern medical diagnosis and treatment， which clarified the positioning and advantages of TCM and provided guidance for clinical and scientific research.

Objective To investigate the correlation between the proportion of peripheral blood follicular T helper cells(Tfh cells)and intracellular interleukin-21(IL-21)content with blue light retinal injury.Methods Brown Norway(BN)rats were randomly divided into 4 groups and were exposed to blue light for 3 hours a day to establish retinal light damage model.According to the duration of illumination,the rats was divided into 0 days(control group),3 days(3 d group),7 days(7 d group)and 14 days(14 d group).The proportion of Tfh cells and content of IL-21 in Tfh cells in peripheral blood of each group was detected by flow cytometry and ELISA sepa-rately after illumination.Electroretinogram(ERG)was used to evaluate retinal function.The changes of fundus in rats were observed by fundus photography.The thickness of outer nuclear layer of retina was analyzed by HE staining.Results After retinal blue light injury,with the extension of illumination time,the proportion of Tfh cells in peripheral blood and intracellular IL-21 content both increased(P<0.05).ERG showed that retinal function decreased after light damage and aggravated with the extension of illumination time,the latency and ampli-tudes of A-wave and B-wave increased and decreased respectively(P<0.05).The retinal fundus of rats showed depigmentation in 3 d,and the retinal vessels became thinner and exudate with the extension of illumination time.HE staining showed that the outer nuclear layer of retina(ONL)became thinner(P<0.05).Correlation analy-sis indicated that the proportion of Tfh cells in peripheral blood and the intracellular IL-21 content could jointly reflect the degree of injury(P<0.000 1),and the proportion of Tfh cells in peripheral blood was negatively corre-lated with ONL thickness and the amplitude of a and b waves,positively correlated with the peak time of a and b waves,(P<0.0001).Conclusion The proportion of Tfh cells in peripheral blood and the intracellular IL-21 content were increased after blue light damage to retina,and were significantly increased with the extension of light time with a certain correlation.

Objective To study the clinical characteristics of outpatients with hand,foot and mouth disease (HFMD) caused by different serotypes of enteroviruses.Methods This was a prospective study.From February 2017 to March 2018,563 outpatients with HFMD were enrolled by systematic sampling in the Department of Infectious Diseases,Henan Children's Hospital.Throat swabs were collected to determine the serotypes via PCR.Demographic,clinical,and laboratory data were collected by standard questionnaire.All cases were followed up twice at 2 and 9 weeks after the initial outpatient visit through telephone interview.A total of 563 cases were enrolled and 555 (98.6％) cases were positive for human enteroviruses,including 338 (60.9％) males.Analyses were stratified by enterovirus serotypes,Chi square test or Fisher's exact test,Rank sum test was used for comparison among different groups.Results The age of 555 cases was 24.2 (16.4,41.3) months.Among them 44.0％ (224 cases) were identified as coxsackievirus (CV)-A6,while 189 cases,35 cases,14 cases and 73 cases were identified as CV-A16,enterovirus (EV)-A71,CV-A10 and other serotypes,respectively.Fever (≥37.5 ℃C) was present in 51.4％(285/555) of laboratory confirmed cases.The proportions of fever in cases of CV-A6 (68.9％(168/244)) and CV-A10 (12/14) were significantly higher than those in cases of CV-A16 (31.7％(60/189),x2=57.344,14.313,both P=0.000),other serotypes (43.8％(32/73),x2=15.101 and 8.242,P=0.000 and 0.004) and EV-A71 (37.1％(13/35),x2=13.506 and 9.441,P=0.000 and 0.002) respectively.There was no significant difference between CV-A6 and CV-A10 in presentation of fever (x2=1.785,P=0.182).There were 359 cases (64.7％) with eruptions in mouth,hands,feet and buttocks.Cases infected with EV-A71 had the highest proportions (74.3％(26/35)) of rash emerging simultaneously in mouth,hands,feet,and buttocks.The proportion in cases of CV-A 16,CV-A6,CVA 10 and other serotype were 73.5％ (139/189),61.9％ (151/244),7/14 and 49.3％ (36/73),respectively.The proportion of rash on other parts of body,such as face,limbs or torso in cases infected with CV-A6 (16.8％ (41/244)) was the higherest and the proportion in cases of CV-A16,EV-A71,CV-A10 or other serotypes were 8.5％(16/189),5.7％(2/35),1/14,6.8％(5/73),respectively.None of these cases developed serious complications.Desquamation occurred in 45.5％ (179/393) cases 7.5 (5.0,9.0) days after disease onset and 13.5％ (53/393) cases showed onychomadesis 31.0 (18.0,33.5) days after disease onset.The proportion of desquamation and onychomadesis associated with CV-A6 (64.2％ (95/148) and 31.8％ (47/148)) was significantly higher than CV-A16 (31.8％ (49/154) and 1.3％ (2/154),x2=33.601 and 52.482,both P=0.000) and other serotypes (38.0％(19/50) and 6.0％(3/50),x2=10.236 and 12.988,P=0.001 and 0.000).Desquamation appeared more in cases of CV-A6 than in cases of CV-A10 (2/11,x2=9.386,P=0.002),with the proportion of onychomadesis higher in CV-A6 than in EV-A71 (3.3％ (1/30),x2=11.088,P=0.001).Conclusion Clinical manifestation such as fever,rash emerging parts,desquamation and onychomadesis are different among outpatient HFMD cases infected with CV-A16,CV-A6,EV-A71,CV-A10 and other enteroviruses.

Objective To research the role of mitochondrial DNA mediate the cultured human retinal pigment epithelium (hRPE) cell apoptosis induced by blue light and the relationship with time.Methods Established the blue light damage model of cultured hRPE cells in vitro with light emitting diode (LED) blue light density of (4.0-±0.5)mW/cm2 adjusted by FL-1D blue light illumination meter,and the illumination time was set as 0,0.5,1,2,4,6,12 and 24 hours,then the cells were grouped according to the illumination time.Immunofluorescence were used to identify the cells;the expressions of caspase-3,cleaved caspase-3,caspase-9,cleaved caspase-9,bax and bcl-2 were detected with Western blot.Quantitative PCR was used to detect the copy number of mitochondrial DNA and PCR was used to detect mitochondrial DNA 4977bp common deletion.Results Immunofluorescence results showed that the RPE65 protein was expressed in the cytoplasm.The expressions of bax were upregulated after illumination for 1 hour,cleaved caspase-3 were upregulated after illumination for 2 hours,caspase-3,caspase-9,cleaved caspase-9 were upregulated after illumination for 4 hours,while the expression of bcl-2 was downregulated after illuminated for 2 hours,with significant differences compared to the normal control group (all at P＜0.05).The copy number of mitochondrial DNA in 0.5,1,2,4,6,12 and 24 hours groups was downregulated,with significant differences compared to the normal control group (all at P＜0.05).The expressions of 4977bp common deletion in 0.5,1,2,4,6,12 and 24 hours groups were increased,with significant differences compared with the normal control group (all at P＜0.05).Conclusions Blue light can cause cell apoptosis,especially mitochondrial apoptosis,in hRPE probably motivated by mitochondrial DNA damage.

Background Light-induced retinal damage results in the damage of retinl pigment epithelial (RPE) cells and therefore affects the pathogenesis and development of age-related macular degeneration (AMD).Studies showed that tissue factor (TF) is overexpressed in oxidative damaged RPE cells and the choroidal neovascularization (CNV) of AMD,speculating that the suppression of TF can prevent the damage of RPE cells and inhibit CNV.Objective This study was conducted to observe the protective effects of TF targeting peptide (TFTP),a new drug of autologous synthesis,on human RPE-cells induced by blue light.Methods Human RPE cells were isolated from donor eye and cultured.Cultured cells were divided into blank control group,model group and TFTP treated group.Light-induced RPE cell damage model was established by exposuring the cells in the blue light of (4.0±-0.5) mW/cm2 for 12 hours in the model group,and different concentrations (10,100,150,200,300 μmol/L) of TF-TP were added into the medium to pretreat the cells for 24 hours and then exposed the cells to the blue light for 12 hours in the TF-TP groups.The cell viability was determined by CCK-8 assay.The morphology and ultrastructure in the cells were observed under the inverted microscope and transmission electron microscope.The apoptosis of the cells was assayed by Hoechst staining.The expressions of TF and apoptosis-related protein bax,bcl-2 in the cells were determined by Western blot.Results CCK-8 assay showed that there was no significant difference in the cell viability among blank control group and different concentrations TF-TP groups (F=2.15,P =0.11).The cell survival rate of blank control group,model group and 150 μmol/L TF-TP group was (100.0±0.00) ％,(43.79±6.55) ％ and (63.45±3.57) ％,and the survial rate was increased in the 150 μmol/L TF-TP group compared with the model group (P =0.00),and 150 μmol/L was detemined as a optimal concentration of TF-TP.A lot of shrinkage,deformation,suspension cells were exhibited under the optical microscope,and decrease of microvilli structure,rupture of mitochondrial cristae and vacuolar degeneration of the cells were found in the model group,and the damage of the cells were evidently lightened in the 150 μ mol/L TF-TP group.The apoptosis rate of the cells were (0.98 ±0.19)％,(9.98 ±0.82) ％ and (5.73 ±0.88) ％ in the blank group,model group and 150 μmol/L TF-TP group,respectively,with a significant difference among the groups (F =206.18,P =0.00),and the apoptosis rate of the cells in the 150 μmol/L TF-TP group was significantly lower than that in the model group (P＜0.05).Compared with the blank control group,the relative expression of bax and TF was obviously increased and that of bcl-2 was decreased in the model group;while the expression of bax and TF was lower,and that of bcl-2 was higher in the 150 μmol/L TF-TP group compared with the model group (all at P ＜ 0.05).Conclusions Pretreation of TF-TP can lessen cell apoptosis and increase cell survival rate and therefore plays a protective role to blue light-induced human RPE cells possibly by inhibiting bax/bcl-2 apoptotic pathways mediated by TF.

Objective To investigate the effects of artesunate (Art) on cell proliferation and apoptosis of human Tenon's capsule fibroblasts (HTFs),and discuss the countermeasures of bleb scarfing in glaucoma.Methods In vitro,HTFs were cultivated and applicated by different concentrations (50 μg · mL-1,100 μg · mL-1,150 μg ·mL-1,200 μg · mL-1) of Art for 48 hours.The effect of Art on cell proliferation was assessed by MTT method.The rate of apoptosis induced by Art was determined by flow cytometry.Western Blot was performed to detect the relative expression levels of Bax and Bcl-2 after Art was treated.Results After treated with Art for 48 hours,compared with blank control group,Art (50 μg · mL-1,100 μg · mL-1,150 μg · mL-1,200 μg · mL-1) group exhibited notable anti-proliferative effect on HTFs with concentration-dependence (all P ＜ 0.05).The results of flow cytometry showed that the apoptosis rates (8.80％ ±0.88％,11.60％ ±0.56％,16.30％ ±1.03％,23.40％ ±1.62％) of HTFs were significantly enhanced with the increase of Art concentration (all P ＜ 0.05).The relative expression levels of Bax were obviously high with the increase of Art concentration,while Bcl-2 levels were significantly low with the increase of Art concentration (all P ＜ 0.05).Conclusion Art can inhibit the proliferation and induce cell apoptosis of HTFs possibly by enhancing the expression of Bax and reducing the expression of Bcl2.Art may be a potential drug in preventing fibrous scar formation after glaucoma filtration surgery.

Mitochondrial DNA (mtDNA) is a genetic effect DNA molecule of double closed loop, and is crucial for cells and their functions. Mitochondria take an active part in physiological activities of retinal pigment epithelium (RPE) cells. The oxidative stress is usually occurred in RPE for its active metabolism, which can lead to mitochondria and mtDNA dam?age. Once mitochondria and mtDNA lesions have not been repaired timely, the lesions can be accumulated, which can cause dysfunctions and damaged-structures of RPE and mitochondria, and can motivate the progression of cell apoptosis. In the end it can result in some ocular related diseases such as aged-related macular degeneration (AMD). This study reviewed the functional relationship between mtDNA and RPE, and repair and detection methods of mtDNA damage.

Background Researches showed that mitochondria and oxidative stress play a crucial role in retinal photochemical injury,but the relationship between the damage of human retinal pigment epithelium (RPE) cell-induced by blue light and light-irradiated time is less studied.Objective The aim of this study was to research the possible mechanism of RPE oxidative damage induced by blue light in vitro.Methods Human RPE cells were isolated from healthy donors and cultured.The cells were divided into the normal control group and the light exposure group.The cells of light exposure group were irradiated using the blue light of (4.0±0.5) mW/cm2 for 0.5,1,2,3,4,5,6,12 and 24 hours,respectively,and the cells of the normal control group were cultured in dark environment.Cellular viability was detected by MTT method,and the ultrastructure change of subcellular organelles in RPE cells was examined under the transmission electron microscope (TEM).The content of reactive oxygen species (ROS) was assayed by flow cytometry for the assessment of oxidative stress reaction.The relative expressions of nicotinamide adenine dinucleotide phosphate (NADPH) mRNA and cyclooxygenase 1 (COX1) mRNA in the cells were detected by real-time fluorescence quantitative PCR to evaluate the mitochondria function.Results The percentages of cellular viability were (100.00±20.00) ％,(95.73±0.89) ％,(94.67±2.56) ％,(84.23±0.16) ％,(78.57±3.09)％,(75.43±2.18)％,(66.13±1.42)％,(53.43±1.91)％ and (47.97±1.36)％ in the normal control group and light exposure for 1-hour,2-hour,3-hour,4-hour,5-hour,6-hour,12-hour and 24-hour groups,respectively,showing a significant difference among the groups (F =172.270,P =0.000),and the percentages of light exposure for the more than 3 hours groups were significantly lower than those of the normal control group (all at P＜ 0.05).The vacuoles-like degeneration,mitochondrial swelling,decreased microvilli were seen under the TEM.The contents of ROS in RPE cells were (14.75±2.49)％,(19.04± 1.02) ％,(22.81 ±3.20)％,(28.75±2.15)％,(33.06±0.96) ％,(40.64±2.11) ％,(48.25±2.50) ％ and (60.44±2.68) ％ in the normal control group and light exposure for 0.5-hour,1-hour,2-hour,3-hour,4-hour,5-hour,6-hour groups,and with significant increases in ROS contents in various light exposure groups compared with the normal control group (all at P＜0.05).The relative expression levels of NAPDH mRNA in the cells were gradually elevated 3 hours after light exposure with the increase of time in comparison with the normal control group (all at P＜0.05),and the relative expression levels of COX1 mRNA in the cells were higher in the light exposure for 2-hour,3-hour,4-hour and 5-hour group compared with the normal control group (all at P＜0.05),and after that the COX1 mRNA levels were gradually declined and were close to the normal level.Conclusions Blue light irradiation for more than 3 hours causes oxidative stress damage of mitochondria in RPE in vitro,and the damage was more obvious after irradiation for 5-6 hours.

Objective To investigate the regulations of Bax , Bcl-2 in the protection of lipoic acid-niacin diad in acrolein-induced apoptosis in ARPE-19 cells. Methods The ARPE-19 cells were cultured in medium containing 10% fetal bovine serum , at 37 ℃ with 5% CO2. The ARPE-19 was transferred to 6-well plate after reaching to 70% confluence. After starvation for 24 h , the cells in 6-well plates were divided into three groups , including the blank control group , the acrolein treatment group with 50 μmol/L acrolein for 24 h , and the protection group with 100 μmol/L lipoic acid-niacin diad for 24 h and with the acrolein for another 24 h. The apoptotic cells were detected by flow cytometry assay , and expressions of Bcl-2 , Bax protein were detected by Western Blot assay. Results The percentages of normal healthy cells were 94.8%, 60.98%, and 91.34% in the blank control group , 50 μmol/L acrolein group and 100 μmol/L diad contained of lipoic acid and niacin group , respectively. The ratios of Bax/Bcl-2 protein expression were 0.293 9, 1.389 2, and 0.555 8 in the blank control group, 50 μmol/L acrolein group and 100 μmol/L diad contained of lipoic acid and niacin group, respectively. Conclusion The protective effect of lipoic acid-niacin diad on acrolein-induced apoptosis in ARPE-19 cell through promoting Bcl-2 expression and inhibiting Bax expression.

OBJECTIVETo observe the influence of flap-on or flap-off Epipolis laser in situ keratomileusis (epi-LASIK) on the release of transforming growth factor-β1 (TGF-β1) in tear fluid and corneal haze formation.

METHODSThirty patients (60 eyes) with myopia underwent epi-LASIK surgery with epithelial flap repositioning (flap-on) in the right eyes and epithelial flap removal (flap-off) in the left eyes. The level of TGF-β1 in tears was measured preoperatively and on days 1, 3, and 7 postoperatively. Corneal haze was graded at 1, 3 and 6 months after surgery.

RESULTSThe mean preoperative spherical equivalent refraction was -4.98∓2.28 D (-2.50 to -7.25 D) in flap-on group and -5.20∓4.02 D (-1.75 to -7.00 D) in flap-off group, showing no significant difference between the two groups (P=0.80). TGF-β1 levels in the tear fluid were similar in the two groups preoperatively (P=0.11) and at 1, 3, and 7 days postoperatively (P=0.55, 0.45, 0.19, respectively). TGF-β1 levels in tears gradually decreased after the first postoperative day in both groups, but were still higher than the preoperative value till the 7th postoperative day. Corneal haze scores in the two groups were similar at 1 month (P=0.98), 3 months (P=0.52), and 6 months (P=0.72) after the operation.

CONCLUSIONFlap-on and flap-off epi-LASIK surgeries do not differ significantly in postoperative TGF-β1 levels in the tear fluid or in the postoperative haze scores. TGF-β1 may play a role in corneal wound healing.

Adult ; Cornea ; surgery ; Epithelium, Corneal ; pathology ; surgery ; Female ; Humans ; Keratomileusis, Laser In Situ ; methods ; Male ; Postoperative Period ; Surgical Flaps ; Tears ; metabolism ; Transforming Growth Factor beta1 ; metabolism ; Young Adult