Objective:To observe the living cells on the surface of intraocular lenses and to compare the cellular numbers and morphology of prestaining to that of poststaining.Methods:Twenty adult pigment rabbit eyes were given posterior chamber intraocular lens implantation.Lenses were extracted and put into 1640 cell cultural liquid immediately on the 1 st,3 rd,7 th,14 th and 28 th day respectively after operatrion.The samples were observed under inverted phase contrast microscope subsequently.Results:We found cells deposited on the surface of lenses from 1 st to 28 th day after operation.These cells contracted and lost three-demensional appearance while its number was less than that of prestaining obviously.Conclusion:A part of cells remove from the surface of lenses during staining;however,we could count the number but could not affirm cellular types which would make error in cellular classification and counting.The fact suggests that we should discover new accurate and reliable methods to avoid mistakes during experiment.

Objective To determine an early and noninvasive predictor of chronic kidney diseaase(CKD),by detecting the correlation between the urinary Col-Ⅳ concentration,the renal tissue and renal lesions in patients with CKD Col-Ⅳ expression.Methods 98 CKD patients proved by renal biopsy were selected and 76 healthy people were enrolled as control.Urinary and serum Col-Ⅳ concentration were examined by enzyme-linked immunosorbent assay(ELISA).Renal Col-Ⅳ expression was detected by immunohistochemistry.Renal injury grade was semiquantitative by CMIAS image analysis system.The association between urinary Col-Ⅳ level and renal Col-Ⅳexpression,as well as renal lesion was analyzed,meanwhile the proteinuria and glomerular filtration rate(GFR)were analysed simultaneously.Results Urinary Col-Ⅳ level in patients with CKD was significantly higher than that in control,however,there was no difference in blood Col-Ⅳ level.Expression of Col-Ⅳ could be found in the sclerosing glomeruli and tubulointerstitial tissue,and Col-Ⅳ expression degree was consistent with lesion degree.Urinary Col-Ⅳ level was correlated positively with renal Col-Ⅳ expression,as well as with density of glomerular basement membrane and interstitial damage.Elevated urinary Col-Ⅳ concentration was observed in patients with mild renal sclerosis prior to the change of GFR.No correlation between urinary Col-Ⅳ concentration and proteinuria was observed.Conclusions These results suggested urinary Col-Ⅳ concentration reflected renal Col-Ⅳ expression and renal sclerosis,and could predict the progression of CKD.

Sixty-four patients with Graves' diseases were divided into methimazole group ( n =30 ) and propylthiouracil group( n =34 ).20 healthy volunteers were used as the control group.The levels of interleukin (IL) -2,IL-6,and TSH receptor antibody (TRAb) were determined by ELISA after serum samples were eollected before treatment and after treatment for 3 and 6 months.The results showed that the general data of two groups were not significantly different before treatment.IL-2 and IL-6 levels at 6 months after treatment were significantly different from the baseline (P＜ 0.05 ),IL-2 being gradually increased while IL-6 decreased with time.The level of IL-6 in methimazole group was lower than that in propylthiouracil group after treatment for 6 months ( P＜ 0.05 ).There was no significant difference in TRAb levels between two groups before treatment while differences became significant at 3 and 6 months of treatment ( all P＜ 0.01 ),being gradually decreased with time. IL-2/IL-6 ratio became significantly greater at 3 and 6 months of treatment compared with that before treatment in the same treatment group( P＜0.05 ) and the ratio in methimazole group was higher than that in propylthiouracil group by 6 months of treatment ( P＜0.05 ).IL-6level was positively correlated with FT3 and FT4 levels,and IL-2 level was negatively correlated with FT3,FT4,and TRAb levels in GD patients before treatment.The correlations of IL-2 and IL-6 with FT3 and FT4 disappeared by 3 and 6 months of treatment.IL-2 and IL-6 levels were related with TRAb level before and after treatment in methimazole group. These results suggest that the immunosuppressive effect of methimazole is more evident than that of propylthiouracil in patients with Graves' disease.

Objective To investigate the expression and significance of TGF‐β receptor Ⅰ /TGF‐β receptor Ⅱ (TGF‐β Ⅰ /TGF‐β Ⅱ) in human skin malignant melanoma A375 cell line .Methods The reverse transcription‐real time polymerase chain reac‐tion (RT‐PCR) and Western blot were utilized to assess the expressions of TGF‐βR Ⅰ /TGF‐‐βR Ⅱ in A375 cell line and human normal melanocytes .Results The expressions of TGF‐βR Ⅰ /TGF‐‐βR Ⅱ mRNA and protein of A375 cells line were significantly lower than those of human normal melanocytes .Conclusion The down‐regulated expression of TGF‐βR in the TGF‐β/Smad signal pathway of human skin malignant melanoma may be one of the pathogenesis of skin malignant melanoma .

Objective To evaluate the effects of problem-based learning (PBL) teaching mode and lecture-based learning (LBL) teaching mode applied in clinical teaching of dermatology in China.Methods All studies on PBL teaching mode and LBL teaching mode applied in clinical teaching of dermatology in China published from 1990 to 2015 were identified by searching in CNKI,VIP database,Wanfang data and so on.Meta-analysis was performed by RevMan 5.3 software.Results Six random controlled trials on 710 clinical students qualified for the meta-analysis according to our criteria.The students in PBL group got significant higher scores than those of the students in LBL group in theoretical scores [WMD=3.75,95％CI=2.58-4.92,P＜0.05],clinical skills tests [WMD=5.27,95％CI=4.60-5.94,P＜0.05] and total scores [WMD=7.93,95％CI=6.49-9.37,P＜0.05].Conclusion PBL teaching mode is an effective mode on teaching of dermatology in China,particularly for theoretical scores and clinical skills,compared with LBL mode.

Objective To observe the hypoglycemic mechanism of total saponins of Momordica charantia (MC) in type 2 diabetes mellitus (DM) rats. Methods Among the selected 60 male Specific Pathogen Free (SPF)rats, 8 were random- ly chosen as control group, while others were fed with high fat and high glucose diet following streptozotocin injection from caudal vein 8 weeks after to construct type 2 DM models. After the DM models were successfully built, rats were then ran- domized into five groups: DM control group (n=8), the metformin group (n=8) and three groups of total saponins of MC with different dosage (n=8 in each group). The total saponins of MC groups include DM rats administrated with total saponins of MC 100, 200, 400 mg/(kg·d) for 8 weeks, the metformin group include DM rats administrated with metformin 50 mg/(kg·d) for 8 weeks. At the end of the experiment, the fasting blood glucose and insulin were examed. At the same time, a part of pan- creas islet, liver and skeletal muscle were preserved. The pancreas islet structure, the hepatic glycogen and Glucose trans- porter 4 (GLUT4) expression were observed by electron microscope, glycogen dyeing and immunoblot respectively. Results After 8 weeks treatment, compared to type 2 DM control group, fasting blood glucose and insulin values in MC groups were reduced more obviously. However, skeletal muscle GLUT4 expression level, insulin granules and hepatic glycogen increased obviously in MC groups. Conclusion Total saponins of MC has hypoglycemic effect. It’s mechanisms maybe include pro- moting the hepatic glycogen synthesis, inhibiting the hepatic glycogen decomposition and promoting insulin sensitivity by in- creasing peripheral tissue’s GLUT4 expression.

Objective To explore the significance of day surgery mode on cataract patients for achieving win-win among patient, hospital and medical insurance.Methods 120 cataract patients (selected from December 2015 to June 2016, in the Ophthalmic Center of Leshan People′s hospital) were included in this retrospective study and day surgery group (n=60) was observation group and hospitalization surgery group was control one (n=60).All cases underwent cataract phacoemulsification and intraocular lens implantation.Treatment process, average day on the waiting list, average preoperative stay, average length of stay, medical cost and patient satisfaction were compared between the two groups.Results Days on the waiting list in day surgery group (1.2 d) reduced 8 days than in hospitalization surgery group (9.2 d).Preoperative stay in day surgery group reduced 1.5 days than in hospitalization surgery group.Length of stay ware reduced by 3 days as compared to the control group.The average medical cost in day surgery group was much less than the control group and the patient satisfaction was much higher.Conclusion The high efficiency of day surgery mode can shorten the length of hospital stay, lessen the cost of medical expense, realize the security management of medical service, so as to provide more convenience, more safety, better quality and cheaper medical service for patients.It achieves social and economic benefits and finally achieves win-win among patient, hospital and medical insurance.

Objective To study the effect of oxymatrine (OMT) combined with radiotherapy on the growth of HO8910 cells of human ovarian cancer in vitro.Methods All human ovarian cancer samples were divided into four groups:control group,treated with culture solution; OMT group,treated with 4 mg/ml OMT; radiotherapy group,treated by 4 Gy radiotherapy; and combination group,firstly treated by 4Gy radiotherapy,and then cultured with 4 mg/ml OMT.Observed the changes of cell morphology by invert microscope at 10,24,48,and 72 hours respectively.Flow cytometry was adopted to detect cell apoptosis.Results ① invert microscope observation showed that compared with the control group,HO8910 cells demonstrated apoptosis of diminution in volume,thickening in cytoplasm,and gathering in nucleus in all the other three groups at 24 and 48 hours.② Flow cytometry showed that apoptosis rate of HO8910 cells and cell population in G1 phase increased in the combination group,which were significantly higher than the other three groups (P＜0.05).Conclusion Both OMT (4 mg/ml)and radiation (4Gy)can induce cell apoptosis,while the combination of them showed better therapeutic results.

BACKGROUND: Vascular endothelial growth factor (VEGF) is a very effective way to make tissue engineer bone vascularization.However, because of expensive and short half-life, VEGF cannot maintain effective concentration in blood after injection. To resolve the problem effectively, gene transfection technique is used in this experiment to transfer human VEGF into seed cells-mesenchymal stem cells (MSCs) of tissue engineer bone and to make it secrete VEGF which could vascularize bone.OBJECTIVE: To explore the possibility of human vascular endothelial growth factor 165 (VEGF165) to transfect rabbit MSCs, and establish the experimental foundation of angiogenesis tissue engineering organization and the treatment of ischemic disorders.DESIGN: Observation control trail.SETTING: First Hospital of Jilin University and Institute of Frontier Medical Sciences of Jilin University.MATERIALS: The experiment was conducted in the Key Laboratory (BSL-2) of Frontier Medical Sciences of Jilin University between June 2003 and August 2004. Health New Zealand white rabbits, 4.0-5.0 months old, weighing 2.5-3.5 kg, half male and half female, were provided by Animal Center of Jilin University. The rabbits were handled under asepsis and anesthetized condition,corresponding to the animal ethical standard. Medicine and reagents: Ham F12 culture media (Gibco, U.S), MTT (Sigma, U.S)PLXSNKDRp-VEGF165 and pcDNA 3.0 vectors were prepared in the present laboratory. ELISA detection kit (Jingmei company,Shenzhen), DH5 α, restriction endonucleases Barn H I, Xhol Ⅰ, Hind Ⅲ, EcoR Ⅰ and standard DNA molecule (Promega,U.S) were also used in this study.METHODS: Rabbits' MSCs were separated and cultivated. The pcDNA 3.0-hVEGF165 expression vector was constructed and identified, pcDNA3.0-VEGF165 eukaryotic expression vector was constructed, the vector was used directly to transfect MSCs. The cultural supernatant then was collected and the soluble protein of human VEGF gene expression was analyzed with ELISA method.The proliferation capability of human umbilical vein endothelial cells (HUVEC) stimulated by the supernatant was measured with MTT methods, untreated MSCs and pcDNA3.0 transfected MSCs were used as control groups.MAIN OUTCOME MEASURES: ① Result of restriction enzyme digestion and DNA sequencing of the recombinant plasmid pcDNA3.0-VEGF165;② the secretion of human VEGF165 proteins of the transfected MSCs analyzed by ABC-ELISA; ③ MTT method was used to detect the effects of MSCs culture supematant transfected with VEGF165 on HUVEC cells proliferation ability.RESULTS: ①Result of restriction enzyme digestion and DNA sequencing of the recombinant plasmid: The constructed plasmid was digested with Hind Ⅲ and XHol Ⅰ, and then two pieces fragments were isolated with agarose gel electrophoresis, which was accordance with expected results. And sequencing results showed that PeDNA3.0-VEGF165 eukaryotic expression vector was successfully constructed. ② ABC-ELISA method: Compared with the control group, concentration of human VEGF protein in the supernatant of the cultured cells increased significantly after the MSCs were transfected with pcDNA3.0-VEGF165 for 24, 48, 72 hours (P＜0.05).③ MTT method was used to detect the effects of MSCs culture supernatant transfected with VEGF165 on HUVEC cells proliferation ability. The results showed MSCs supematant transfected with VEGF165 (2%, 4%,8%, 16%, and 32%) had statistical significance in promoting HUVEC cells proliferation rate compared with the normal control (P＜0.05).CONCLUSION: Human VEGF gene can be successfully transfected into MSCs and expressed effectively.

Objective To investigate the effect of the eye-acupuncture therapy on serum TNF-α levels in rats with cerebral ischemia-reperfusion injury and the related mechanism. Methods Healthy SD rats were randomly divided into four groups: normal group, sham-operated control group, ischemia-reperfusion model group and eye-acupuncture group according to body weight. Sham-operated control group, ischemia-reperfusion model group and eye-acupuncture group were divided into the 3h group, the 24h group and the 72h group, a total of 10 groups (n=8). To establish the rat model of cerebral ischemia-reperfusion by suture method in ischemia-reperfusion model group and eye-acupuncture group. Eye-acupuncture was separately started immediately after reperfusion and at 30 min before sampling in the 3h eye-acupuncture group, besides, eye-acupuncture was separately taken every 12h in the 24h eye-acupuncture group and in the 72h eye-acupuncture group. The rats in normal group were not treated, those in sham-operated control group were inserted the fishing thread 1cm, and the others were identical with those in ischemia-reperfusion model group. At 3h, 24h, 72h after reperfusion, the neurophysical behaviours were accessed by ZeaLonga neurophysical impairment marks in ischemia-reperfusion model group and eye-acupuncture group. The method of ELISA was taken to detect the change of serum TNF-α levels in rats after the eye-acupuncture therapy. Results Compared with the ischemia-reperfusion model group, the neurologic impairment score of the eye-acupuncture group decreased obviously; The levels of serum TNF-αin ischemia-reperfusion model group at 3 h, 24 h, 72 h respectively were(76.803±18.325)pg/ml、(85.511±13.334)pg/ml、(86.831±9.232)pg/ml. The level of serum TNF-α in normal group was(24.304±6.511)pg/ml. The level of serum TNF-α in Sham-operated control group at 3 h, 24 h, 72 h respectively were(24.928±3.792)pg/ml,(27.533±5.362)pg/ml,(29.366±5.874)pg/ml. Ischemia-reperfusion model group compare with normal group and sham-operated control group, the difference were significant(P＜0.01). The levels of serum TNF-α in rats after the eye-acupuncture therapy at 3 h, 24 h, 72 h respectively were(40.185±3.335)pg/ml, (48.523±7.687)pg/ml, (51.611± 6.403)pg/ml. Compared with ischemia-reperfusion model group, the levels of serum TNF-α were strongly reduced(P＜0.01). Conclusion The eye-acupuncture therapy could play a role in improving cerebral ischemia-reperfusion injury evidently and the mechanism was related to its reducing serum TNF-α levels.