This study investigated the effect of etoposide, an anticancer chemotherapy drug, on B7-H1 expression in retinoblastoma (Rb) cells and the role of miR-513a-5p in the process. Rb cells were divided into control and etoposide groups. In the etoposide group, cells were treated with etoposide at different concentrations (2.5, 5, 10, 20 and 40 μg/mL) for 24 h. Those given no treatment of etopside served as controls. Reverse transcription polymerase chain reaction (RT-PCR), fluorescence quantitative PCR and flow cytometry were performed to measure the mRNA and protein expression of B7-H1 in Rb cells. The mRNA expression of miR-513a-5p in Rb cells before and after etoposide treatment was also detected by fluorescence quantitative PCR. The miR-513a-5p mimics and the miR-513a-5p inhibitor were transfected into Rb cells separately, and fluorescence quantitative PCR and flow cytometry were used to detect the effect of the miR-513a-5p mimics or inhibitor on B7-H1 expression. TargetScan5.2 was employed to predict the miR-513a-5p binding sites in the 3'-untranslated region of B7-H1 mRNA. Luciferase reporter plasmids carrying this site were prepared and transfected into Rb cells and luciferase activity analyzed. The results showed that etoposide stimulated the mRNA and protein expression of B7-H1 in Rb cells, which reached a maximal level after treatment with 5 μg/mL etoposide (P<0.05). However, miR-513a-5p expression was decreased in Rb cells after etoposide treatment. When the miR-513a-5p inhibitor was added, B7-H1 expression was increased with the concentration of the miR-513a-5p inhibitor (P<0.05). Moreover, B7-H1 expression was decreased gradually with the concentration of the miR-513a-5p mimics increased (P<0.01). Additionally, the miR-513a-5p mimics were found to inhibit the luciferase activity. It was concluded that etoposide can promote B7-H1 expression in Rb cells, which may be associated with chemoresistance. The promoting effect of etoposide on B7-H1 expression can be reversed by miR-513a-5p mimics. MiR-513a-5p inhibits the mRNA and protein expression of B7-H1 via binding to the 3'-UTR of B7-H1 mRNA.

OBJECTIVETo observe the effect of Jianpi Yangzheng Xiaozheng Recipe (JYXR) on the tumor inhibition rate of subcutaneous transplanted tumor gastric cancer cell line MGC-803 in BALB/c nude mice, and to study its molecular mechanism of apoptosis and autophagy.

METHODSGastric cancer cell line MGC-803 was subcutaneously inoculated to nude mice for preparing transplanted gastric cancer models. Totally 32 BALB/c nude mice were randomly divided into 4 groups according to random digit table, i.e., the negative control group, the positive control group, the high dose JYXR group, the low dose JYXR group, 8 in each group. Normal saline was administered to mice in the negative control group by gastrogavage. 5-fluorouracil (5-Fu) at 2. 5 mg/kg was administered to mice in the positive control group by gastrogavage. JYXR at 85 and 43 g/kg was administered to mice in the high dose JYXR group and the low dose JYXR group by gastrogavage, once per day for 10 successive days. The effect of JYXR on the tumor inhibition rate of subcutaneous transplanted tumor was observed. Effects of JYXR on gene expression levels of Bax, Bcl-2, Fas, Cyclin D1, Cyclin D2, and Cyclin D3 in transplanted tumor were observed by real-time PCR. Effects of JYXR on protein expression levels of Procaspase-3, Procaspase-8, Procaspase-9, cleaved-PARP, Beclin-1, and LC3B were detected using Western blot.

RESULTS(1) Compared with the negative control group, the tumor weight was obviously reduced in the rest three groups (P <0. 05). The tumor weight was higher in the high dose JYXR group and the low dose JYXR group than in the positive control group (P <0. 05). (2) Results of RT-PCR indicated that, compared with the negative control group, expression levels of Bax were up-regulated, but expression levels of Bcl-2, Cyclin D1, Cyclin D2, and Cyclin D3 were down-regulated in the positive control group and JYXR groups (P <0. 05). The expression level of Fas was up-regulated in the positive control group and the high dose JYXR group (P <0. 05). Compared with the positive control group, expression levels of Fas, and Bax were all down-regulated, but expression levels of Bcl-2, Cyclin D2, and Cyclin D3 were all up-regulated in the high dose JYXR group and the low dose JYXR group (all P <0. 05). The expression level of Cyclin D1 was down-regulated in the high dose JYXR group, but it was up-regulated in the low dose JYXR group ( both P <0. 05). (3) Results of Western blot showed, compared with the negative control group, expression levels of Procaspase-3, Procaspase-8, and Procaspase-9 were down-regulated, but expression levels of cleaved-PARP, Beclin-1, and LC3B II were up-regulated in the high dose JYXR group and the low dose JYXR group (all P <0.05). Compared with the negative control group, expression levels of Procaspase-3, Procaspase-8, Procaspase-9, and LC3B II were down-regulated, but expression levels of cleaved-PARP, Beclin-1, and LC3B I were up-regulated in the positive control group (all P <0. 05).

CONCLUSIONSJYXR showed significant inhibition on subcutaneous transplanted tumor gastric cancer cell line MGC-803 in BALB/c nude mice. Its mechanism might be associated with activating apoptosis and autophagy correlated factors.

Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; Autophagy ; drug effects ; Caspase 3 ; metabolism ; Caspase 8 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Chemistry, Pharmaceutical ; methods ; standards ; Cyclin D1 ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Fluorouracil ; Humans ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Stomach Neoplasms

The medium and process parameters were optimized in batchaminoglycoside antibiotic JI-20A fennentation. The optimal medium consists mainly of comstarch 60g/L, peanut meal 30g/L, com slurry 8g/L, maltose 10g/L, inorganic compound and amylase moderate , methionin 1g/L and cobalt chloride 6?g/mL. It was significant to adjust medium pH after autoclaving and oxygenate the fennentation medium for the cell growth and JI-20A product.

Objective To investigate the expression of c-Myc in gastric mucosa associated lymphoid tissue (MALT) lymphoma and its implications on prognosis.Methods The clinical data of 79 patients hospitalized in our hospital from 2009 to 2015 were retrospectively analyzed.In these patients,38 patients were low-grade MALT lymphoma,20 patients were high-grade MALT lymphoma,21 patients were diffused large B cell lymphoma (DLBCL).Real-time PCR was used to detect the levels of c-Myc expression in gastric MALT tissues and pair-matched adjacent normal tissues.The relationship between the expression of c-Myc and prognosis of patients was evaluated combing with the clinical data.Results Compared with the normal tissues,the expression levels of c-Myc protein were 15.7％ (6/38),25％ (5/20) and 28.5％ (6/21)in patients with low-grade MALT lymphoma,high-grade MALT lymphoma,and DLBCL.The relative expression levels of cMyc mRNA were gradually elevated in low-grade MALT lymphoma,high-grade MALT lymphoma and DLBCL.The tumor size and depth of invasion can influence the expression level of c-Myc.Survival analysis found that the overall survival rates and relapse-free survival rates were lower in patients with c-Myc positive expression than those of patients with negative expression (P ＜ 0.05).Conclusion C-Myc plays a key role in the malignant transformation of gastric MALT lymphoma.

Objective To monitor the quality changes of iodized salt and analyze its impact factor in Chongqing between 2001 and 2009. Methods Salt samples were collected according to the east, west, south,north and center locations in iodized salt production, wholesale and household sectors. Two units in iodized salt production and wholesale segment were sampled from north, south, east and west places and only 1 unit was sampled from the central place. Nine samples were collected every month in each place. If the place had less than 9 units, and then taken all the units. About resident household, 2 townships were sampled from north, south, east and west places, and 1 township was sampled from the central place, then 20 samples were collected from each township. Iodine content was detected by oxidation-reduction assay. The index of mean iodine, qualified rate from factories and wholesale, coverage rate and taking rate of qualified iodized salt in residents were calculated.Significance was analyzed by trend test, analysis of variance and X2 test. Results The qualified rate of iodized salt from the manufacturers was 92.9%(13/14) in 2001 and the rate was 100.0% each year from 2002 to 2009. The qualified rates of iodized salt from the wholesale were 88.7%(282/318) - 99.8%(431/432). The rates of 2001 and 2002 were lower than that of other years(X2 = 4.98 - 45.69, all P< 0.05 or < 0.01). The coverage rate and taking rate of qualified iodized salt in residents were 94.2% (11 154/11 841 ) - 98.9% ( 14 061/14 217), 83.5% (9 887/11 841 ) -95.8% (13 449/14 039), respectively. The rates showed an increasing tendency (F = 9.27, 26.39, all P < 0.05).The districts(counties) with qualified iodized salt consumption rate > 90% kept increasing. The mean iodine from the manufacturers and wholesale were 29.71 - 36.25, and 31.26 - 36.13 mg/kg, respectively. The iodine level showed a descending trend(F = 35.45, 140.59, all P < 0.01 ). The mean iodine level from the inhabitants were 28.84 - 30.98 mg/kg which remained stable (F = 3.05, P > 0.05 ). The iodine level from manufacturers, wholesale to inhabitants showed an descending trend(F = 38.46 - 671.23, all P < 0.01 ). Conclusions The surveillance results of iodized salt shows an increasing tendency in quality of iodized salt, eoverage rate and taking rate of qualified iodized salt. Factors that affect the quality of iodized salt is that the enterprise does not add iodine to salt strictly by the standard.

The method was established for determination of geniposidic acid, chlorogenic acid, pinoresinoldiglucoside, which are three kinds of constituents of Eucommia ulmoides absorbed into the blood components. LC-MS/MS technique was applied to determine the blood components of the bloodstream after administration of E. ulmoides extract. At the same time, HPLC was used for detection of the ingredients content of the blood sample from 23 batches of E. ulmoides. The results showed that geniposidic acid, chlorogenic acid and pinoresinoldiglucoside are prototype into the blood in rats after oral administration of E. ulmoides extract, The linear range of geniposidic acid, chlorogenic acid and pinoresinoldiglucoside was good, and the average recoveries geniposidic acid, chlorogenic acid and pinoresinoldiglucoside were 98.69%, 100.8% and 98.39%, respectively. The method is simple and feasible with good reproducibility.
Animals ; Chlorogenic Acid ; blood ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; Eucommiaceae ; chemistry ; Glycosides ; blood ; Iridoid Glucosides ; blood ; Lignans ; blood ; Male ; Plasma ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tandem Mass Spectrometry

OBJECTIVEThe biological safety of a new developed silicone rubber for inflatable silastic prosthesis (SRISP) was evaluated.

METHODSFollowing the GB/T 16886.10-2005 standard, YY/T 0127.13-2009 standard, and GB/T 16886.11- 2011 standard, samples were prepared and tested by animal experiments, such as guinea pig maximization test, oral mucous membrane irritation test, and short-term systemic toxicity test (oral route).

RESULTSNo obvious erythema and edema in the guinea pig abdominal skin were observed after 24, 48, and 72 h of stimulating touch, thus indicating that SRISP does not cause potential skin sensitivity. No local response to SRISP was found, and the visual observation and pathological findings of oral mucosa were normal and similar to that of the control group. Therefore, SRISP had no irritation response to oral mucosa. No clinical signs of toxicity were observed in rats, and no significant differences in weight and weight relative growth rate between extract group and blank control group (P > 0.05) were found. Thus, SRISP had no short-term systemic toxicity.

CONCLUSIONThese results indicated that SRISP met the requirement of biomedical materials and had good bio- security.

Animals ; Biocompatible Materials ; Cosmetics ; Dimethylpolysiloxanes ; Guinea Pigs ; Prostheses and Implants ; Rats ; Silicone Elastomers ; Toxicity Tests

Objective Toinvestigatetheriskoflong-termrecurrenceofstrokeanditspredictorsin youngpatientswithischemicstroke/transientischemicattack(TIA).Methods Theclinicaldataofthe consecutive young patients (18-45 years)with ischemic stroke/TIA (within 2 weeks after onset)admitted to the department of neurology in the First Affiliated Hospital of Sun Yat-Sen University between August 2008 and July 2013 were enrolled prospectively. All patients were regularly followed up for a long time (The patients were followed up at the 1 st,6 th,and 12 th month after onset;then they were followed up once for every 6 months)in order to investigate stroke recurrence. The Kaplan-Meier curves were used to analyze the cumulative stroke recurrence rate of all patients. The last contact time for patients lost to follow was used as censored data to be enrolled in the analysis. The univariate analysis of the related risk factors for stroke recurrence using Log-rank test. Multivariate Cox proportional hazard regression was used to detect the related risk factors associated with stroke recurrence (adjusting for age and sex). The variables of the results of Log-ranktestP≤0.1wasselectedandenrolledinthemultivariateregressionanalysis.Results Atotalof 312 patients were enrolled in the analysis,including 294 with ischemic stroke and 18 with TIA. Their mean follow-up time was 34 ± 19 months. Thirty-four patients had recurrent stroke,including 23 with ischemic stroke,7 with TIA,and 4 with cerebral hemorrhage. The cumulative recurrence rates of stroke at 1 ,3 , and 6 years after onset were 6. 2%,10. 3%,and 16. 4%,respectively. The results of multivariate Cox proportional hazards regression analysis showed that hypertension (risk ratio [RR]2. 159;95% confidence interval [CI]1. 048-4. 447,P=0. 037)and cardioembolism (RR,2. 869;95%CI 1. 119-7. 357,P=0.028)weretheindependentpredictorsforstrokerecurrence.Conclusion Theoverall6-yearriskof recurrent stroke is not high in the Chinese young patients with ischemic stroke/TIA,but the risk of stroke recurrence is relatively higher in the first year. Hypertension and cardioembolism are the potential predictors of stroke recurrence;therefore,attention should be paid in clinical practice.

Objective To investigate the risk factors, diagnostic evaluation, etiology, and treatment in young patients with ischemic stroke. Methods The clinical data of young patients (age range 18 - 45 years) with ischemic stroke (within 2 weeks of stroke onset) admitted to the Stroke Center, the First Affiliated Hospital of Sun Yat-sen University from August 2008 to July 2013 were registered prospectively. Results A total of 300 patients were enroled, their age was 37. 8 ± 6. 8 years. There were 227 males (75. 7% ). The age of 84 patients (28. 0% ) was ≤35 years. The most common risk factors were smoking (43. 3% ), hypertension (38. 7% ), and hypercholesterolemia (38. 0% ). The positive result detection rates were lower in the antinuclear antibody (8/278), anticardiolipin antibody (34/250), 24-h Holter monitoring (2/60 ), and transthoracic echocardiography (38/232). According to the MRI findings, the detection rate of the white matter changes, old infarcts, single acute infarcts, and multiple acute cerebral infarcts were 40/282, 77/282, 145/282, and 137/282, respectively. The stroke subtypes of TOAST: large artery atherosclerosis (26. 7% ), smal artery occlusion (18. 0% ), cardioembolism (10. 0% ), other determined etiology (23. 0% ), and undetermined etiology (22. 3% ). Large artery atherosclerosis was mainly involved in anterior circulation (69/80) and intracranial arteries (75/80). In other definite causes, cerebral artery dissection was most common (36/69), and other causes included moyamoya disease (7/69) and infectious vasculitis (7/69), etc. The average length of hospital stay was 15. 4 d. The mean National Institutes of Health Stroke Scale score on admission was 7. 0, and the mean modified Rankin scale score at discharge was 2. 5. The complication rate during hospitalization was 9. 7% . 80. 3% and 48. 3% of patients received anti-platelet drugs and statins therapy during hospitalization.Conclusions This study used a prospective single-center method. It conducted a comprehensive analysis of risk factors, diagnostic evaluation, treatment, and etiology in current Chinese young patients with ischemic stroke. Its research data wil provide useful information for establishing a diagnostic strategy of high performance cost ratio, in-depth understanding of its pathophysiological mechanisms, and improving treatment strategies in Chinese young patients with ischemic stroke.

Objective To assess the clinical application value of phleboplasties combined with microvascular anastomotic device in venous anastomosis with diameter discrepancy in head and neck defects reconstruction.Methods Sixty-six pairs of veins with significant diameter discrepancy were anastomosed in head and neck reconstructive surgeries with free flaps.Forty of them were anastomosed with microvascular anastomotic device (the coupler group) after phleboplasties including lateral incision, Y-T enlargement and wedge excision while the other 26 pairs of veins were conventionally sutured (the sutured group).Diameter of each vein, anastomosis time, post-operative vascular crisis, flap survival and complications related to the microvascular anastomotic device were recorded.Results The average anastomosis time of the coupler group was (4.78 ± 1.14) min for lateral incision, (5.16 ± 2.07) min for Y-T enlargement and (11.09 ± 3.21) min for wedge excision, and all of them were significantly shorter than that of the sutured group.In the sutured group, all flaps survived except for 2 veins with poor blood flow were cut and re-anastomosed during the operation;1 flap with venous crisis within 72 hours after the operations was explored and replaced with the pectoralis major myocutaneous flap.All veins in the coupler group were successfully anastomosed in a single coupling procedure without anastomotic impatency, blood leak, vessel tearing and ring shedding.No vascular crisis occurred postoperatively.One patient underwent cervical haematoma 5 hours after the operation, and the flap blood supply was unaffected after the haematoma was removed.All flaps in the coupler group survived completely.Patients in both two groups were followed up 6 to 18 months.All flaps healed perfectly and no obvious surgical complications or microvascular anastomotic device rejection happened.Conclusion When anastomoses are carried out using microvascular anastomotic device between veins of different size, phleboplasties including lateral incision., Y-T enlargement and wedge excision can not only reduce the size discrepancy and the anastomosis time, but also ease the difficulty level and guarantee the patency of the venous anastomoses.Wedge excision enjoys the advantage of haemodynamics, and obstruction of venous reflux hardly occurred for size reduction.It should be considered preferentially when external jugular veins are used as the anastomotic vein of the recipient sites in head and neck reconstruction.