Background and purpose:Smac/DIABLO was the only apoptosis-related protein that could inhibit IAPs directly and simultaneously.The four amino-residual AVPI(Ala-Val-Pro-lie)in its N-terminal was the very important domain that could stimulate apoptosis.This study investigated the effect of synthetic Smac peptide (SmacN7) on chemotherapy sensitivity of bladder cancer cells.Methods:SmacN7 penetratin peptide was synthesized and delivered into T24 cells.MTT assay was adopted to evaluate the viability of T24 cells induced by low-dosage of MMC. Flow cytometry was applied to analyze the proportion of apoptosis and Western blot was used to detect the expression of XIAP and caspase-3;The activity of caspase-3 was measured and the effect of SmacN7 combined with MMC on T24 cell lines was also determined.Results:SmacN7 penetratin peptide could successfully interact with endogenous XIAP and increase the proportions of apoptosis of T24 cell lines induced by low-dosage of MMC in a dose-and time- dependent manner.An obvious down-regulation of XIAP expression and up-regulation of caspase-3 was identified by Western blot.The activity of caspase-3 in experimental group was significantly increased as compared with that in the control group;Combining the treatment with SmacN7 penetratin peptide,the viability of T24 cells decreased to 55% and 72.7% in 24 hrs and 48 hrs respectively.Conclusion:SmacN7 penetratin peptide could act as a cell-permeable IAP inhibitor,inhibit the proliferation,induce apoptosis and enhance the chemo-sensitivity of bladder cancer cells to MMC. When combined with chemotherapy,it may be a very promising strategy for bladder cancer therapy.

Objective:To study the biocompatibility of fibrin sealant (FS) and human corneal fibroblasts (HCFs) obtained by small incision lenticule extraction (SMILE).Methods:The human corneal stromal tissues were selected from corneal stromal lens in 24 eyes of 12 patients underwent SMILE in the First Affiliated Hospital of Guangxi Medical University from March to April 2018.HCFs were isolated and cultured in vitro within 1 hour after the corneal stromal lens were extracted and the growth status of HCFs on FS surface was observed.HCFs were divided into 2-fold leaching solution group and normal control group, and the cells in the two groups were treated with 2-fold leaching solution or complete medium according to grouping, respectively.The apoptosis of HCFs in the two groups was observed by acridine orange (AO)/ethidium bromide (EB) double staining.The proliferation of HCFs in the two groups was assayed by methyl thiazolyl tetrazolium (MTT) method.HCFs in logarithmic phase were divided into 2-fold leaching solution group, normal control group, and the cells were treated with 2-fold leaching solution or complete medium according to grouping, respectively.In addition, a blank control group without HCFs was also set and treated with complete medium.The absorbance value and relative growth rate of HCFs in the three groups were compared.HCFs in logarithmic phase were divided into 1-fold leaching solution group, 2-fold leaching solution group and normal control group, and the cells were treated with 1-fold leaching solution, 2-fold leaching solution or complete medium culture according to grouping, respectively.The apoptosis of HCFs in the three groups was compared by Annexin V-FITC/PI flow cytometry, and the cytotoxicity of the three groups was graded.Written informed consent was obtained from each patient before the operation.The study protocol adhered to the Declaration of Helsinki and was approved by the Ethics Committee of the First Affiliated Hospital of Guangxi Medical University (No.2018[022]). Results:HCFs grew well on FS surface and the morphology was normal.MTT assay showed that HCFs in the 2-fold leaching solution group and the normal control group had a similar proliferation tendency, and the toxicity index of HCFs in the 2-fold leaching solution group was graded 0-1 at 0-72 hours after changing solution.After AO/EB staining, the HCFs in the 2-fold leaching solution group and the normal control group were normal, and only a small amount of early apoptotic cells were observed.Flow cytometry showed that the apoptosis rates of the normal control group, once leaching solution group and the double leaching solution group were (4.96±1.09)%, (3.66±1.35)% and (2.88±0.66)%, respectively, with no significant difference among them ( F=2.89, P=0.13). Conclusions:FS has no cytotoxicity and has good biocompatibility with HCFs in vitro.

BACKGROUND: Bistort rhizome is also named as caoheche, which is characterized by clearing heat, relieving convulsion, regulating damp and reducing swelling. Additionally, its water extract is characterized by antiarrhythmia and central inhibition; however, analgesia should be studied further.OBJECTIVE: To observe analgesic effect of polygonum bistorta L. Water extract, and compare with morphine and amidazofen.DESIGN: Completely randomized digital table and randomized controlled animal study.SETTING: Department of Pharmacology, Gannan Medical College.MATERIALS: The experiment was carried out in the Laboratory of Scientific Center of Gannan Medical College from March to May 2004. ① A total of 150 healthy adult Kunming mice were used in the 4 independent experiments. ② Medicines: Polygonum bistorta L. Water extract (Department of Phytochemistry, Shenyang Pharmaceutical University; batch number: 2003061001); morphine hydrochloride solution (Shenyang First Pharmaceutical Factory; batch number: 000305); naloxone hydrochloride solution (Yanqiao pharmaceutical Co. Ltd.; batch number: 20021109).METHODS: ① Effect of polygonum bistorta L. Water extract on twisting-body reaction of mice induced by acetic acid: Forty mice were randomly divided into 4 groups: saline group, 0.10 and 0.15 mg/g polygonum bistorta L. Water extract groups and amidazofen group with 10 in each group. All mice were intraperitoneally injected with 0.02 mL/g saline,0.10 and 0.15 mg/g polygonum bistorta L. Water extract solution and 0.10 mg/g amidazofen, respectively. Fifteen minutes later, mice were intraperitoneally injected with 6 g/L 0.01 mL/g acetic acid glacial to record times of twisting-body reaction within 15 minutes. ② Effect of polygonum bistorta L. Water extract on pain response of mice induced by hot-plate test: Forty female mice were randomly divided into 4 groups:saline group, 0.10 and 0.15 polygonum bistorta L. Water extract groups and morphine group with 10 in each group. All mice were intraperitoneally injected with 0.02 mL/g saline, 0.10 and 0.15 mg/g polygonum bistorta L. Water extract solution and 0.01 mg/g morphine solution, respectively. GJ-8402 hot-plate pain response threshold detector was used in this study; pain response temperature was (55.0±0.5) ℃; pain response after licking hindfoot was regarded as reactive marker; latency of pain response threshold was within 60 s. Pain response was measured at 15, 30 and 60 minutes after administration with hot-plate test. ③ Effect of morphine, naloxone and polygonum bistorta L. Water extract on pain response of mice induced by hot-plate test: Thirty female mice were randomly divided into 3 groups: saline group, naloxone+morphine group and naloxone+polygonum bistorta L. Water extract group with 10 in each group. All mice were intraperitoneally injected with 0.02 mL/g saline, 0.004 mg/g naloxone solution+0.01 mg/g morphine solution and 0.004 mg/g naloxone solution+0.15 mg/g polygonum bistorta L. Water extract solution, respectivelu. Pain response was measured at 15, 30 and 60 minutes after administration with hot-plate test. ④ Effect of polygonum bistorta L. Water extract on pain response of mice induced by electric stimulation: Forty mice were randomly divided into 4 groups with 10 in each group. All mice were intraperitoneally injected with 0.02 mL/g saline, 0.10 and 0.15 mg/g polygonum bistorta L. Water extract and 1 g/L morphine, respectively. Pain response was measured at 20, 35, 50 and 70 minutes after administration with electric stimulus method.MAIN OUTCOME MEASURES: ① Times of twisting-body reaction; ②duration of pain response induced by hot-plate test; ③ analgesic rate induced by electric stimulation.RESULTS: All 150 healthy adult Kunming mice were involved in the final analysis. ① Times of twisting-body reaction: At 15 inutes after administration, times of twisting-body reaction were less in 0.10 and 0.15 mg/g polygonum bistorta L. Water extract group and amidazofen group than those in saline group [(15.1±11.1), (8.0±6.5), (6.3±3.2), (54.1±20.2) times, t=3.532-3.681, P ＜ 0.01]. ② Duration of pain response induced by hot-plate test:At 15, 30 and 60 minutes after administration, durations of pain response induced by hot-plate test were longer in 0.10 and 0.15 mg/g polygonum bistorta L. Water extract group and morphine group than those in saline group (t=2.676-3.683, P ＜ 0.05-0.01). ③ Duration of pain response was longer in naloxone + polygonum bistorta L. Water extract group than that in saline group at each time point after administration (t=2.676-3.563, P＜ 0.05-0.01); however, duration in naloxone + morphine group was close to that in saline group (P ＞ 0.05). ④ Analgesic rate induced by electric stimulation: At 20, 35, 50 and 70 minutes after administration, analgesic rate induced by electric stimulation was higher in 0.10 and 0.15 mg/g polygonum bistorta L. Water extract group and morphine group than that in saline group (t=3.455-3.634, P ＜ 0.01).CONCLUSION: ① Polygonum bistorta L. Water extract has the obviously analgesic effect, whose intensity is close to that of amidazofen and morphine. ② Naloxone, an opiate receptor antagonist, can resist analgesic effect of morphine but not that of polygonum bistorta L. Water extract. This suggests that analgesic effect of polygonum bistorta L. Water extract dose not react through exciting opiate receptor.

One new dicyclopeptide cyclo-(L-N-methyl Glu-L-N-methyl Glu) (1), together with one new natural dicyclopeptide cyclo-(L-methyl Glu ester-L-methyl Glu ester) (2), and two known dicyclopeptides cyclo-(L-methyl Glu ester-L-Glu) (3), and cyclo-(L-Glu-L-Glu) (4), were isolated from the aerial parts of Dianthus chinensis L. Their structures were determined by spectroscopic analyses and chemical methods.
Dianthus ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry

An experimental model of rhegmatogenous retinal detachment (RRD) in rabbits was established to simulate the pathophysiologic condition of human RRD. 24 rabbits were randomly divided into 3 groups and underwent vitrectomy with a vitrector and/or retinotomy with a Charles flute needle, with 12 in group I (vitrectomy and retinotomy), 7 in group I (retinotomy) and 5 in group III (vitrectomy). All animals underwent follow-up examinations with direct and indirect ophthalmoscopy and fundus photography 12 h and day 1, 3, 5, 7, 10, 14, 21, and 28 after the procedure(s). Retinal changes were recorded. As a result, 10 RRDs were successfully established in group I. Direct and indirect ophthalmoscopy and fundus photography demonstrated typical features of RRD. No RRD developed in group II and III. It was concluded that the experimental rhegmatogenous retinal detachment produced in a rabbit model after vitrectomy with retinotomy in this study was a convenient and reliable one. This RRD model mimicked the typical pathophysiological changes in humans.
*Disease Models, Animal ; Random Allocation ; Retina/surgery ; *Retinal Detachment ; Vitrectomy

Background A main cause of visual impairment in proliferative diabetic retinopathy (PDR) is vitreous hemorrhage and retinal detachment due to contraction of fibrovascular membrane.To explore the pathogenic mechanism of fibrovascular membrane is a new target for the prevention and management of PDR.Objective This study was to determine the change in expression of vascular endothelial growth factor (VEGF),connective tissue growth factor(CTGF) and pigment epithelium derived factor(PEDF) in the proliferative membranes of patients with PDR after intravitreal injection of avastin,an anti-VEGF agent.Methods This study was approved by the Medical Ethic Committee of Tianjin Medical College,and written informed consent was obtained from each patient before enrollment.A prospective randomized-controlled study was designed.Twenty-six eyes of 24 patients with PDR scheduled for surgery were enrolled from January to June,2008 in Tianjin Medical College Eye Hospital.The patients were randomized into the simple vitrectomy group and avastin injection combined with vitrectomy group,with matched gender,age and disease duration.1.25 mg (0.05 ml) of avastin was intravitreally injected prior to surgery,and vitrectomy was performed 10 days after injection in the avastin injection combined with vitrectomy group,and only vitrectomy was given in the simple vitrectomy group.Preretinal membrane was collected during the surgery.Expression of VEGF,CTGF and PEDF in the preretinal membranes was assayed by immunochemistry.Results VEGF,CTGF and PEDF were expressed in the cytoplasm.The rate of VEGF expression in the preretinal membranes was 30.77％ in the avastin injection combined with vitrectomy group,showing a significant reduction in comparison with the simple vitrectomy group(100.00％)(U =4.000,P＜0.01).The rate of expression CTGF was remarkable elevated in the avastin injection combined with vitrectomy group compared with the simple vitrectomy group (92.31％ vs.62.54％)(U=7.500,P=0.048).However,no significant difference was found in the expression rate of PEDF between the two groups(100.00％ vs.92.31％) (U =65.500,P =0.299).Conclusions The results suggest that intravitreal injection of anti-VEGF drugs resulted in the decrease of VEGF expression and increased CTGF expression in proliferative membranes from patients with PDR.

A sensitive, rapid, simple and economical ultra-performance liquid chromatography–tandem mass spectro-metric method (UPLC–MS/MS) was developed and validated for simultaneous determination of imatinib, dasatinib and nilotinib in human plasma using gliquidone as internal standard (IS). Liquid-liquid extraction method with ethyl acetate was used for sample pre-treatment. The separation was performed on an Xtimate Phenyl column using isocratic mobile phase consisting of A (aqueous phase: 0.15% formic acid and 0.05% ammonium acetate)and B(organic phase:acetonitrile)(A:B=40:60,v/v).The flow rate was 0.25 mL/min and the total run time was 6 min. The multiple reaction monitoring (MRM) transitions, m/z 494.5→394.5 for imatinib, 488.7→401.5 for dasatinib, 530.7→289.5 for nilotinib and 528.5→403.4 for IS, were chosen to achieve high selectivity in the simultaneous analyses. The method exhibited great improvement in sensitivity and good linearity over the concentration range of 2.6–5250.0 ng/mL for imatinib, 2.0–490.0 ng/mL for dasatinib,and 2.4–4700.0 ng/mL for nilotinib.The method showed acceptable results on sensitivity,specificity, recovery, precision, accuracy and stability tests. This UPLC–MS/MS assay was successfully used for human plasma samples analysis and no significant differences were found in imatinib steady-state trough concentra-tions among the SLC22A5?1889T>C or SLCO1B3 699G>A genotypes(P>0.05).This validated method can provide support for clinical therapeutic drug monitoring and pharmacokinetic investigations of these three tyrosine kinase inhibitors(TKIs).

Objective To study the anti-tumor activity of quercetin in NB4 leukemia cells and the roles of PI3K/Akt,bcl-2,and Bax on the quercetin-induced apoptosis,and to investigate the potential underlying mechanism.Methods MTT assay was used to monitor cell proliferation,Hoechst 33258 fluorescent staining and flow cytometry were employed to detect apoptosis in NB4 cells.Western blot was used to detect the expression changes of related proteins in quercetin treated NB4 cells.Confocal laser microscopy was used to test the distributional variation of Akt between cytoplasm and nucleus.Results Quercetin significantly inhibited the NB4 cell proliferation in a dose-and time-dependent manner (20-160 μ mol/L).In addition,treated by 20,40 80 μmol/L quercetin,the rates of apoptosis were (9.25±0.11) ％,(20.83±2.10) ％and (41.43±2.90) ％,there were statistical difference compared with blank cells (t were 4.14,6.56 and 7.02,all P ＜ 0.05).This was concentration dependent and accompanied by morphological changes characteristic of apoptosis.Further,quercetin induced a G~M arrest,which might account for its cytotoxic effects.Quercetin decreased PI3k/Akt expression and caused an inhibition of the anti-apoptotic protein bcl-2,while increasing the expression of Bax.Quercetin had no effects on total Akt,but it promoted Akt translocation from cell nucleus to the cytoplasm (F =15.12,P ＜ 0.05).Conclusion Quercetin induces the leukemia NB4 cell apoptosis by affecting multiple signal pathways and plays a strong anti-leukemia effect.In addition,our results suggest that PI3K/Akt pathway could be a novel target for the leukemia chemotherapy.

Facing current situation of integrative medicine (IM), authors put forward that clinical and diagnosis program of IM could be carried out from clinical path, pathogenesis, treatment theory and philosophy, and so on, but with different integration degrees. Meanwhile, formulation of concrete program should be disease-targetedly set up, and adjusted from person to person, from place to place, from time to time. As for settled IM program , authors could evaluate it from whether Chinese medicine and Western medicine have formed complementary, synergistic, excitatory actions, and toxicity attenuation; whether more problems could be solved in efficacy, safety, practicability, and economy than previous single mode.
Critical Pathways ; Integrative Medicine ; trends ; Medicine, Chinese Traditional ; trends

Improving medical students' calculation ability in statistics has become the focus and difficulty of medical statistics course teaching, and its application relies heavily on statistical calculation software. In order to explore a new teaching approach which combined the advantages of traditional method and web-based calculation, we intended to build a web computing platform applying the Browser/Server (B/S) mode based on the campus network, and to revise the current syllabus of medical statistics, as well as to create a virtual web-lab containing a typical case library. Practice has proved that the new mode effec-tively improved the practice capability of students and changed the traditional teacher-centered teaching approach.