Objective To investigate the effect of hyperthermic intraperitoneal perfusion and cisplatinon apoptosis in human ovarian cancer cells.Methods Two human ovarian cancer cells (OVCAR-3,A2780) were divided into control group,cisplatin group,hyperthermia group and thermo-chemotherapy group;microscopy was used to observe the morphological changes of the four groups;AO/GV stain and flow cytometry(FCM) was used to analyze cell apoptosis;Apoptosis related genes caspase7,caspase8 and Bax in ovarian cancer cells were detected by fluorescence quantitative PCR.Results Inverted microscopy observeed that the ovarian cancer cells retracted and suspended partially in the cisplatin group and hyperthermia group,especially in the thermo-chemotherapy group.After AO/GV staining,the apoptotic cells were increased in the cisplatin group and hyperthermia group compared with the control group,and the thermo-chemotherapy group was more than cisplatin group and hyperthermia group.FCM results indicated that the proportion of cells apoptosis were higher in the cisplatin group and hyperthermiagroup,the thermo-chemotherapy group is the higher than the all other groups(P＜0.05).q-PCR results showed that in the thermo-treatment group the expression of pro-apoptotic genes,including caspase3,caspase6,caspase7,caspase8,caspase9,Bax,Bak and Bid,was significantly higher than other groups,apoptosis inhibitory gennes,such as Bcl-2,Bcl-xL,Mcl-1,c-FLIP,was significantly decreased than the others.Conclusions Cisplatin plus hyperthermia can promote the apoptosis in ovarian cancer cells.

The aim of this study is to explore the anti-HSV activity of P.vulgaris polysaccharides and gels in vitro and in vivo with the provision of scientific evidence for the further research of anti-HSV new drugs.Plaque reduction assay was adopted to determine the IC50 value of P.vulgaris polysaccharides on HSV-1 and HSV-2 in vitro.In addition,HSV-1 (skin) and HSV-2 (vulva) infected guinea-pig models were established for assessing the anti-HSV activity of polysaccharides and gels in vivo,and infected lesion degree,number of papulovesicle,typical lesion score and the DNA copy numbers of HSV-1 and HSV-2 viruses in the lesion tissue were taken as the indexes.It was found that the activities of HSV1 and HSV-2 viruses was inhibited by P.vulgaris polysaccharides in vitro,while HSV induced skin lesions in the guineapigs were ameliorated by P.vulgaris gel,exerting an anti-HSV action.In conclusion,it was demonstrated that P.vulgaris polysaccharides and gels performed an anti-HSV action both in vitro and in vivo with the hidden value of developing antiHSV agents.

OBJECTIVE:To study the percutaneous permeability of methoxypsoralen liposomal gel METHODS:The preparations were applied on the rat skin in vitro and in vivo with common gel and Mingbailing tincture as control,and the drug concentrations in the receiver,skin,blood and other organs were determined by RP-HPLC RESULTS:In vitro experiment,liposomal gel showed the lowest percutaneous permeation rate and the highest retention in skin In vivo studies illuminated that the skin depositions of drug encapsulated in liposome was(4 37?0 91) and (3 36?0 58) times those of the common gel and tincture respectively CONCLUSION:Methoxypsoralen liposomal gel showed an evident targeting effect on skin

OBJECTIVE:To measure the concentrations of levofloxacin in aqueous humor after a single eye dripping of drug and calculate the pharmacokinetic parameters.METHODS:32 rabbits were divided into 8 groups.The rabbit eyeballs were enucleated to make a collection of aqueous humor at 0.125,0.25,0.5,0.75,1.0,2.0,3.0 and 4.0 hours after dripping.All measurements were performed with HPLC.RESULTS:In aqueous humor,Tmax:0.940h,Cmax:0.592?g/ml,T1/2(Ka):0.479h,T1/2 (Ke)0.919h.By using 3p87 pharmacokinetic program,the concentration-time curves were fitted as one-compartment op_en model.CONCLUSION:All the concentrations of levofloxacin at different times after dripping in aqueous humor exceeded the minimum inhibitory concentration for 90%(MIC90)of most bacterial pathogens.These results show that levofloxacin lactate eye-drops may be an effective antimicrobial agent for the treatment and prevention of intraocular infections as well as other ocular infections.

Objective To explore the therapeutic effect influencing factors of intra-aortic balloon pump(IABP) support during cardiac surgery perioperative period and countermeasures.Methods Clinical data of 42 patients using IABP in the cardiac surgery was analyzed retrospectively.The patients were classified into 2 groups:living group (32 patients) and died group (10 patients).The time of IABP implantation,preoperative cardiac function,the duration of cardiopulmonary bypass(CPB) and aorta block,mean arterial pressure (MAP) and inotropic score (IS) before using IABP and postoperative complications were recorded and compared.Results In died group,2 patients were used IABP preoperatively,1 patient was used intraoperatively,7 patients were used postoperatively,8 patients were in NYHA cardiac function class Ⅲ or Ⅳ preoperative,the duration of CPB and aorta block were (144.43 ± 49.03),(97.29 ± 39.99)min respectively,MAP and IS before using IABP were (57.34 ±7.25) mm Hg (1 mm Hg =0.133 kPa) and (28.22 ±17.72) scores,IABP time was (86.00 ±52.31) min.Compared with living group,all above comparisons showed significant difference [(100.43 ± 35.03) min,(60.45 ± 20.55) min,(69.34 ± 8.05)mm Hg,(10.82 ± 15.75) scores,(49.00 ± 25.23) min] (P ＜ 0.05).Postoperative complications in died group included 7 patients of ventilator dependency,6 patients of acute renal failure,5 patients of refractory metabolic acidosis,2 patients of malignant arrhythmia,1 patient of acute myocardial infarction,significantly higher than those in living group (0,4,2,0,0) (P ＜ 0.05).Conclusions Erroneously choose the timing of using IABP,poor preoperative cardiac function,prolonging CPB and aorta block time,MAP less than 60 mm Hg and high dose positive inotropic agent before using IABP,and postoperative complications are main influencing factors for clinical outcomes of IABP in cardiac surgery.Reasonably choosing adaptive indication and timing of IABP,preventing and treating postoperative complications effectively can improve effects of IABP in cardiac surgery.

Objective To investigate the significance of renal tubular dysfunction in patients with refractoriness nephrotic syndrome(RNS)and the effect of interference treatment of Valsartan (VAL).Methods 79 cases of RNS and 68 healthy controls were recruited into the study. The 79 patients of RNS were divided randomly into the VAL group and the dipyridam group. On the basis of routine therapy, the VAL group was given VAL(80mg/d),and the dipyridam group taken dipyridam (150mg/d)orally for 12 weeks The glomerular tubular function(u-RBP,α1-MG,β2-MG,mAlb,NAG)were detected and the pathologic changes of tubukinterstitium were observed by using the methods of ELISA, biochemistry and scoring of the pathologic damage of tubulointerstitium before and after 12 weeks of VAL treatment in all of the cases. Results Urinary RBP,α1-MG,β2-MG,mAlb and NAG in all patients with RNS was evidently higher than that in healthy controls(P＜0.01).Those had positive correlations with damages of tubulointerstitium(r=0.436,0.626,0.499,0.668,0.657,P＜0.01).The interference outcome displayed that the excretion rates of urinary series of protein after oral use VAL in treatment group were markedly lower, while the control group had no distinct change. Conclusion There were various injury of tubulointerstitium and the disfiguration of renal tubular function in all cases with RNS. Damages of renal tubular function had positive correlation with tubulointemtitium injury and renal globular injury. Interference treatment with ARB in patients with RNS could improve renal tubular function, which is of great significance in delaying the progress of RNS.

OBJECTIVE:To evaluate the biological compatibility and clinical therapeutic effect of the medical sterilized bone wax.METHODS:Embedding test and pathological test as well as hemolytic test were performed by using rabbits as test animal.12531case-times were investigated in respect to the therapeutic effect,prognosis and satisfactory rate.RESULTS:The embedded bone wax was coated by connective tissue and has not been absorbed without surrounding inflammation,edema or necrosis.The bone wax could not be absorbed after stanching bleeding of the bone broken surface.No hemolytic phenomena were observed.The wound healing obtained a satisfactory rate of99.1%.CONCLUSION:The sterilized medical bone wax has good biological compatibility and is safe and effective to stanch bleeding.It is convenient to use due to its singer dose sterilized packaging.

OBJECTIVE:To prepare the sterilized medical bone wax and to establish the standard of quality control.METHODS:The bone wax was identified with chemical approach and the quality of bone wax was evaluated by saponification value.RESULTS:The bone wax was appropriate in formula,feasible in preparing technique and satisfactory in therapeutic efficacy with a satisfication rate of 98%.CONCLUSION:There are no obvious differences between the bone wax developed by our hospital and imported bone wax in quality,therefore the prepared bone wax can take the place of imported products.

Objective: To detect the mutation of epidermal growth factor receptor (EGFR) gene in peripheral blood of non-small cell lung cancer (NSCLC) patients in Yunnan area with Super-ARMS, and to explore its correlation with clinicopathological characteristics. Methods: A total of 222 blood samples from patients with NSCLC were collected between January 2017 to December 2018 in the Molecular Diagnostic Center of Yunnan Cancer Hospital. The EGFR gene mutation in peripheral blood samples was detected by SuperARMS, and the relationship between EGFR gene mutation and clinicopathological features was analyzed. Meanwhile, the independent risk factors influencing EFGR mutation were also analyzed. Results: In the peripheral blood of 222 NSCLC patients, there were 81 cases (36.5%) with EGFR gene mutation. Among them, exon 19 deletion and L858R gene point mutation were the most common (75.3% of total mutation); female patients had a higher mutation rate than male patients (45.9% vs 27.0%); patients <60 years old had a higher incidence of mutation than patients≥60 years old (43.2% vs 28.8%) (P<0.05 or P<0.01); moreover, patients with no history of smoking, no history of radical surgery, adenocarcinoma, advanced stage and no history of chemotherapy had higher incidence of EGFR mutation (43.9% vs 21.6%, 39.2% vs 21.2%, 43.9% vs 4.8%, 39.7% vs 23.3% and 44.0% vs 23.5%) (P<0.05 or P<0.01). Multivariate logistic analysis showed that young, no smoking history, adenocarcinoma and no surgical history were independent risk factors for EGFR gene mutation (all P<0.01). Conclusion: In the peripheral blood of patients with NSCLC in Yunnan, the mutation rate of EGFR gene is higher in patients with age<60 years old, adenocarcinoma and non-smoking. Super-ARMS method is more sensitive in the detection of EGFR mutation in peripheral blood of lung cancer patients.

ObjectiveFrom the changes of expression of cold-inducible RNA-binding protein (CIRP) in rats with traumatic brain injury under mild hypothermia treatment with Shenfu decoction as a subsidiary, to speculate the mechanism of protective effect of the decoction on the injury.Methods Ninety Sprague-Dawley (SD) rats were divided into three groups by random number table: non-transfection control group, adenovirus mediated immune flourescent reverse transcription virus group (blank AD5-GFP transfection group) and adenovirus mediated immune flourescent reverse transcription virus carrying CIRP silent expression gene group (AD5-GFP-CIRP-SiRNA transfection group), 30 rats in each groups. Then, each group was subdivided into three subgroups: model group, traditional Chinese medicine (TCM) low and high dose groups, 10 rats in each subgroup. After the mild hypothermia treatment for 48 hours, in the TCM low dose group and high dose group, a dose of TCM 1 mL/kg and 5 mL/kg was injected via a tail vein into the rat respectively, while in the model group, 1 mL/kg normal saline was injected into the same vein, once a day for consecutive 2 days in all the groups. Before modeling in the blank AD5-GFP transfection group and AD5-GFP-CIRP-SiRNA transfection group, virus transfection models were reproduced at first by one-time intrathecal injection of 0.1 mL AD5-GFP and 0.1 mL (1×1010 pfu/mL) AD5-GFP-CIRP-SiRNA virus vector respectively, and in model group, 0.1 mL normal saline was given. The rat cortex, hippocampus and hypothalamus part were collected, the brain cell apoptosis was detected by transferase-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL), the CIRP mRNA expression in the cortex, hippocampus and hypothalamus part was measured by reverse transcription-polymerase chain reaction (RT-PCR), the protein expressions of rat sarcoma protein Raf, Ras, extracellular signal-regulated kinase (ERK), phosphorylation ERK (p-ERK), mitogen activated protein kinase (MEK), p-MEK were determined by Western Blot.Results The brain tissue cell apoptosis indexes (AI) in the cortex, hippocampus and hypothalamus part in TCM low, high dose group of non-transfection control and blank AD5-GFP transfection group were lower than those in model group, and the expressions of CIRP mRNA were higher than those in model group, there were no significant differences in AI and CIRP mRNA in the cortex, hippocampus and hypothalamus between model, TCM low and high dose groups of AD5-GFP-CIRP-SiRNA transfection group, but AI was significantly higher and CIRP mRNA was significantly lower than that in corresponding subgroups of AD5-GFP transfection control group and blank AD5-GFP transfection group. Western Blot detection showed that: Raf/Ras, p-MEK/MEK protein expressions revealed no statistical significant differences in different parts of each group (allP > 0.05), the p-ERK/ERK protein expression in the cortex, hippocampus, and hypothalamus part was significantly lower in TCM low and high dose group than that in the model group of non-transfection control group and blank AD5-GFP transfection group, the degree of descent in the TCM high dose group being more significant (the cortex: non-transfection control group was 7.2±1.0 vs. 15.3±1.8, AD5-GFP transfection group was 8.1±0.7 vs. 16.2±1.5; hippocampus part: non-transfection control group was 6.6±0.8 vs. 14.7±2.0, AD5-GFP transfection group was 6.8±1.0 vs. 14.9±1.3; hypothalamus part: non-transfection control group was 9.4±1.1 vs. 12.7±1.7, AD5-GFP transfection group was 10.6±1.3 vs. 9.4±1.1, allP < 0.05). There were no significant statistical differences in p-ERK/ERK protein expression in above brain parts between AD5-GFP-CIRP-SiRNA transfection subgroups (allP > 0.05).Conclusions The Shenfu decoction used in rats with brain trauma under treatment of mild hypothermia is possibly by promoting CIRP over-expression, lowering ERK expression and inhibiting the initiation of signal transduction of the secondary transcription factor phosphorylation, thereby the neural cell apoptosis is decreased and play a subsidiary role of anti-apoptosis of mild hypothermia.