OBJECTIVE:To establish a diazotize solution-spectrophotometry for the determination of bilirubin in zhenhuang oral liquid.METHODS:The solution to be determined was added with diazotize solution and the absorbability was detected at a detective wavelength of533nm.RESULTS:The calibration curve was linear for bilirubin at the range of1.07～8.56?g/ml(r=0.9999).The average recovery of loading sample was100.2%(RSD=0.73%).CONCLUSION:The method is simple,reproducible,cost-effective and suitable for the determination and quality control of bilirubin in zhenhuang oral liquid.

Objective To explore the effects of TMPRSS2 promoted suicide gene CD-5-FC combined with phytochemicals on the proliferation and apoptosis of prostate cancer (PCa) specific cell-line 22RV1.Methods From March 2016 to October 2016 TMPRSS2-VISA-CD/UPRT'vector was used to deliver pro-drug 5-FC (5-Fluorocytosine) in to PCa specific cell-line 22RV1.Transfection effect was verified by Western-blotting.5-FC,5-FU (5-Fluorouracil) and four phytochemicals epigallocatechin gallate (ECGC),Genistein,Daidzein,and Equol were selected in this study.MTS assay was performed to select dosages,which can kill 10％-40％ of 22RV1 cells,from their pre-set drug concentrations,respectively.According to their select concentrations,either 5-FC plasmid or 5-FU in combination with one of four phytochemicals were added in 22RV1 cells in orders,and then cultured them together.Besides,these four phytochemicals were added in 22RV1 cells individually regarding their selecting concentrations and cultured at the same time.Cell viability was detected by MTS assay on 24hrs,48hrs and 72hrs.Comparing the cell killing rates between the combination groups and each single drug groups on cell-line 22RV1.Results TMPRSS2-VISA-CD/UPRT'plasmid with 5-FC was transfected on PCa specific cell-line 22RV1 successfully.The most obvious transfection occurred on 48hrs.The selected concentration of the pro-drug 5-FC were 83.32 μmol/L and 833.20 μmol/L,5-FU were 76.87 mol/L and 768.70 μmol/L.The concentrations of four phytochemical agents were 2.00 μmol/L,10.00 μmol/L and 20.00 μmol/L.In term of the cell killing numbers,results showed that the combination treatment of 83.32 μmol/L 5-FC/ 76.87μmol/L 5-FU and 10.00 μmol/L/20.00 μmol/L four phytochemicals compared with 10.00 μmol/L/20.00 μmol/L single treatment of four phytochemicals,respectively,there was no significant difference (P ＞0.05),while the combination treatment of 833.20 μmol/L 5-FC/768.70 μ mol/L 5-FU and 2.00 μmol/L/10.00 μmol/L four phytochemicals compared with 2.00 μ mol/L/10.00 μmol/L single treatment of four phytochemicals,respectively,there was a significant difference (P ＜ 0.05).In addition,this study proved that 83.32 μmol/L 5-FC and 76.87 μmol/L 5-FU,833.20 μmol/L 5-FC and 768.70 μmol/L 5-FU reached the same cell killing effect,and there was no statistical difference (P ＞ 0.05).Thus the suicide gene transfection was successful,and its function was the same as the effect of cytotoxic drugs.Conclusions This study has proved that this plasmid with 5-FC suicide gene system can be successfully and efficiently transfected into PCa cells 22RV1.As the pro-drug of 5-FU,5-FC got similar treatment effect with 5-FU,and inhibited cell proliferation.There was no synergistic reaction of enhancing cell apoptosis after combined phytochemical drugs EGCG,Genistein,Daidzen,Equol with suicide gene therapy on 22RV1 cell-line.

Objective to investigate the emotional state and childhood trauma among users of amphetaminetype-stimulants under compulsory seclusion rehabilitation in Shaoxing.Methods A cross-sectional design was conducted on 140 users of ATS in Shaoxing compulsory isolation drug rehabilitation center,collecting general information using homemade questionnaires.Using the Patient Health Questionnaire (PHQ-9),Generalized Anxiety Disorder Scale (GAD-7) and Mood Disorder Questionnaire(MDQ) to investigate the emotional state of users of ATS.The Childhood Trauma Questionnaire (CTQ) was applied to explore the childhood trauma experiences of users of ATS.Results Among the 140 users of ATS,84 (60％) users had emotional problems,24 users (17.14％) of anxiety disorder,22 users (15.72％) of depression,27 users (19.28％) of depression and anxiety disorder.66 users (47.1％)experienced at least one childhood trauma.The most common childhood trauma is the physical neglect,60 users (42.9％),followed by emotional neglect in 23 users(16.4％),again sexual abuse in 18 users (12.9％),emotional abuse and physical abuse is relatively fewer.Conclusion Among users of ATS under compulsory seclusion rehabilitation,emotional problems is prevalent.Proper emotional intervention and treatment in users of ATS would promote their mental health,thereby reducing the re-use of ATS,better return to society life.

Type 2 diabetes(T2DM) is the collective result of insulin resistance in both muscle and liver andβ cell function failure. Traditional pharmaceutical therapy fails in ceasing progressive decline in β cell function. Bariatric surgery has been widely accepted as an efficient measure to treat diabetes. However, the prevalent standards mainly based on body mass index can not meet the demand for promoting remission rate. Related factors should be included in prediction of post-operative remission, like end-point setting, T2DM status, biochemical index, and operative method etc. The combination of these factors can be used to benefit more patients with diabetes.

Choroidal thickness and its relevance with retinal disease has been widely studied in recent years,as choroid is an important source of retina blood supply.Diabetic retinopathy (DR) studies have been focused on retinal vascular injury and related circulatory disorders for a long time,however recent studies have found that choroidal blood vessels are also affected by diabetes,including the thickness changes.The choroidal thickness of diabetic patients is thinner than normal.There is no conclusive conclusion about choroidal thickness and the severity of DR lesions,as the choroid thickness results are determined by multiple factors,including diseases,ocular conditions (choroidal vascular status,ocular axial length,refractory errors and other eye parameters),systemic factors (age and sex) and measurement methods.Therefore,it is necessary to calibrate the ocular and extra-ocular factors affecting choroidal thickness when trying to further clarify the relationship between the choroidal thickness and DR.

Objective To investigate the clinical efficacy of surgical treatment for stage Ⅳ gastric cancer after conversion therapy.Methods The retrospective cohort study was conducted.The clinicopathological data of 50 stage Ⅳ gastric cancer patients who were admitted to the First Affiliated Hospital of Dalian Medical University between January 2012 and June 2016 were collected.All the 50 patients who were diagnosed with single distal metastasis underwent chemotherapy.After chemotherapy,21 patients with operation indication receiving gastrectomy (R0 or R1 resection) were allocated into the conversion surgery group and 29 without operation indication continuing to chemotherapy were allocated into the chenotherapy group.Patients received S-1 + oxaliplatin or S-1 + docetaxel regimen.Patients underwent open distal or total gastrectomy.Observation indicators:(1) response assessment of chemotherapy (complete remission,partial remission,stable disease and progressive disease),grading of of adverse reactions;(2) intra-and post-operative situations of conversion surgery group:operation procedures,intraoperative situations (operation time,volume of blood loss,number of lymph node dissected and surgical margin) and postoperative situations (complications and duration of hospital stay);(3)comparison of follow-up and survival between the 2 groups;(4) prognostic factors analysis affecting stage Ⅳ gastric cancer patients.Follow-up using outpatient examination,correspondence and telephone interview was performed to detect postoperative survival of patients up to September 2016.Survival time was from operation to the last follow-up or death.Measurement data with normal distribution were represented as x±s.Comparison of count data and univariate analysis were done using the chi-square test.Ordinal data were analyzed by the nonparametric test.The survival rate was calculated by the Kaplan-Meier method,and the Log-rank test was used for survival analysis.Multivariate analysis was done using the COX regression model.Results (1) Response assessment of chemotherapy:of 50 patients,24 received S-1 + oxaliplatin regimen and 26 received S-1 + docetaxel regimen.Twenty-one patients in the conversion surgery group underwent chemotherapy,with negative peritoneal metastasis,N2 and below of lymph node metastasis (No.16 lymph node disappeared or reduced),invasive depth ＜T4b and narrowing or disappeared hepatic metastasis.A median preoperative chemotherapy cycle was 4.2 cycles (range,2.0-9.0 cycles).Chemotherapy reaction of 21 patients:15 had partial remission and 6 had stable disease.Twenty-nine patients without operation indication in the chemotherapy group didn't receive surgery.The median cycle of first-line chemotherapy was 5.5 cycles (range,2.0-10.0 cycles).Chemotherapy reaction of 29 patients:13 had partial remission,11 had stable disease and 5 patient had progressive disease.Chemotherapy adverse reactions of 50 patients:26 had reduced white blood cells (WBCs),including 6 with grade Ⅲ-Ⅳ of adverse reactions;29 had reduced neutrophils,including 12 with grade Ⅲ-Ⅳ of adverse reactions;18 had anemia,including 6 with grade Ⅲ-Ⅳ of adverse reactions;12 had reduced platelets,including 2 with grade Ⅲ-Ⅳ of adverse reactions;27 had apositia,including 5 with grade Ⅲ-Ⅳ of adverse reactions;7 had stomatitis;9 had diarrhea;3 had elevated serum creatinin;4 had hand-foot syndrome;3 had abnormal sensory nerve.There was no chemotherapy-related death.(2) Intra-and post-operative situations of conversion surgery group:of 21 patients in the conversion surgery group,8 underwent radical total gastrectomy + D3 lymph node dissection,6 underwent radical distal gastrectomy + D3 lymph node dissection and 7 underwent radical distal gastrectomy + D2 lymph node dissection (including 4 combined with resection of hepatic metastatic tumors and 1 combined with radiofrequency ablation of hepatic metastatic tumor).Operation time,volume of blood loss,number of lymph node dissected and recovery time of gastrointestinal function of 21 patients were (216±31)minutes,(128±52)mL,31±8 and (3.0± 0.7)days,respectively.There were 17 patients receiving R0 resection and 4 receiving R1 resection (3 with positive gastric margin and 1 with positive hepatic margin).There was no death.Seven of 21 patients with complications were cured by conservative treatment,including 2 with pancreatic fistula,1 with intra-abdominal hemorrhage,1 with intestinal obstruction,1 with pneumonia,1 with intra-abdominal infection and 1 with wound infection.Duration of hospital stay of 21 patients was (13.0±3.0) days.(3) Comparison of follow-up and survival between the 2 groups:50 patients were followed up for 6-46 months,with a median time of 24 months.The 3-year cumulative survival rates in the conversion surgery and chemotherapy groups were respectively 33.3％ and 6.9％,with a statistically significant difference between the 2 groups (x2 =7.678,P＜0.05).Results of further analysis showed that R0 resection of 17 patients and R1 resection of 4 patients in the conversion surgery group were respecgtively (25.3±2.8)months and (8.3±0.9)months,with a statistically significant difference between the 2 groups (X2=16.242,P＜0.05).(4) Prognostic factors analysis affecting stage Ⅳ gastric cancer patients:results of univariate analysis showed that T stage,N stage,response assessment of chemotherapy,surgery after chemotherapy and degree of tumor radical resection were related factors affecting prognosis of stage Ⅳ gastric cancer patients (X2 =5.288,12.645,25.581,8.372,12.001,P＜0.05).Results of multivariate analysis showed that R1 resection after conversion therapy was an independent risk factor affecting prognosis of stage Ⅳ gastric cancer patients (HR=14.021,95％ confidence interval:1.928-10.938,P＜0.05).Conclusion Radical resection after conversion therapy can increase survival rate of stage Ⅳ gastric cancer patients,and R1 resection after conversion therapy is an independent risk factor affecting poor prognosis of stage Ⅳ gastric cancer patients.

Angiogenesis Inducing Agents ; metabolism ; Angiogenesis Inhibitors ; therapeutic use ; Animals ; Antibodies, Monoclonal ; therapeutic use ; Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; therapeutic use ; Bevacizumab ; Drug Delivery Systems ; Endostatins ; therapeutic use ; Humans ; Neoplasms ; drug therapy ; Neovascularization, Pathologic ; drug therapy ; Receptors, Vascular Endothelial Growth Factor ; metabolism ; physiology ; Vascular Endothelial Growth Factors ; metabolism ; physiology

Objective To establish an evaluation index system for access to ophthalmic surgery so as to standardize the access. Method Collect 14 ophthalmologists' recommendations on access criteria for ophthalmic surgery in two rounds of questionnaire survey by Delphi method. Results The return rate of rounds of consultation questionnaire was 93.33%, and the average expert authority coefficient was 0.78.Nine ophthalmic surgery access indicators were established. The coordination of expert coefficient was 0.521 after two consultation. It had significant difference ( P＜0.01 ). Conclusion The ophthalmic surgery access indicator system is highly credible and allows quantitative assessment of ophthalmic surgery from various aspects. It is more demanding for ophthalmologist, and gets more involvement of patients in the evaluation of ophthalmic surgery access. It could contribute to scientific and rational decision-making on access for ophthalmic surgery.

Objective To observe the ultrastructure of dimorphic Penicillium marneffeiisolates from wild bamboo rats (Rhizomys pruinosus) in Guangxi region as well as from a patient with penieilliosis marneffei,and to compare their biological characteristics and anti-oxidative mechanisms.Methods Two Penicillium marneffei strains,including one isolated from wild bamboo rats (Rhizomys pruinosus) in Guangxi region and one from a patient with penicilliosis marneffei,were cultured with or without the presence of 2.0 mmol/L hydrogen peroxide in potato dextrose agar (PDA) at 25 ℃ and in brain-heart infusion (BHI) broth at 37℃ for seven days.The shape of colony and growth of both strains were observed.Light microscopy was carried out to study the morphology,and transmission electron microscopy to observe the ultrastructure,of both isolates.Results Ater incubation with hydrogen peroxide,there was a slowdown in the growth of both Penicillium marneffei isolates at both mycelial phase and yeast phase,with an increase in the production of pigment at mycelial phase at 25℃.No obvious changes were observed at 37 ℃ in the morphology of either the clinical isolate or the bamboo rat isolate when cultured with hydrogen peroxide compared with those cultured without hydrogen peroxide.Light microscopy showed attenuated spore formation by the clinical isolate when cultured at 25 ℃ with hydrogen peroxide,crenation of both isolates when cultured at 37 ℃ with hydrogen peroxide.Under a transmission electron microscope,the mycelial cells of both isolates exhibited smooth cell walls,intact cell membranes,with nuclei,mitochondria,endoplasmic reticulum,lipid body,vacuoles of various sizes in the cytoplasm at 25 ℃,and even microbodies at 37 ℃,when cultured without the presence of hydrogen peroxide.After incubation with hydrogen peroxide,the cell wall of both isolates became incomplete with defects in some areas and uneven thickness,the cell membrane discontinuous with shrinkages and projections,and the cytoplasm was inhomogeneous with obvious phagocytosis and numerous phagocytic vacuoles.Conclusions The clinical and bamboo rat isolates of Penicillium marneffei experience different biological and morphological changes under oxidative stress,hinting differences in antioxidative mechanism between them.

Objective To explore clinical effect of surgical treatment of displaced intra-articular calcaneal fractures via modified lateral L-shaped incision.Methods From January 2005 to October 2011,133patients (143 feet) with displaced intra-articular calcaneal fractures,including 125 males and 8 females,aged from 19 years to 65 years (average,43.2 years),underwent open reduction and internal fixation via modified lateral L-shaped incision.There were 56 cases of left calcaneal fractures,67 cases of right calcaneal fractures,and 10 cases of bilateral calcaneal fractures,and all of them were closed fracture.According to Sanders classification,15 feet were classified as type Ⅱ,107 type Ⅲ,and 21 type Ⅳ.The American Orthopaedic Foot and Ankle Society (AOFAS) ankle-hindfoot scale were used to access outcomes.Results One hundred and twenty five patients (135 feet) were followed up for 12 to 28 months (average,18.5months).All fractures healed after an average of 13 weeks (range,8-16 weeks).There were on nerve injury or osteomyelitis.Incision dehiscence occurred in 4 feet,which healed after removing the plate.Subtalar joint traumatic arthritis occurred in 17 feet,with walking pain.Collapse of articular surface occurred in 5 feet after weight-bearing.According to AOFAS ankle-hindfoot scale,excellent result was got in 94 cases,good in 29 cases,fair in 14 cases,poor in 6 cases; and the excellent and good rate was 92.9％.Conclusion Open reduction and internal fixation via modified lateral L-shaped incision for treating displaced intra-articular calcaneal fractures can obtain satisfactory results,but the skilled surgical techniques are needed.