Objective: To establish a method for determination of methone and pulegone in Herba Schizonepetae. Methods: The contents of menthone and pulegone were determined by GC (equipped with FID) with HP-5 fused capillary column (5% phenyl methyl siloxane 30m?0.32mm?0.25?m) after the samples were extracted by the solvents. Results: The linear ranges were 0.002-5.0g/L (r=0.9999) for menthone and 0.002-5.0g/L (r=0.9996) for pulegone, the recoveries of menthone and pulygone were 96.30%-103.9% and 95.7%-102.5%, respectively. Conclusion: The method was simple and accurate, which could be applied to the determination of menthone and pulegone in Herba Schizonepetae.

Objective To explore the necessity of inferior vena cava(IVC) filter implantation during catheter directed thrombolysis (CDT) for acute deep vein thrombosis (DVT). Method Ninety-three patients with acute DVT were reviewed from Nov. 2006 to Dec. 2008. There were 35 men and 58 women, age averaging at (60 ±29) year old, treated with CDT followed by percutaneous transluminal angioplasty ( PTA). The course of DVT was from 5 h ～ 15 d (6. 28 ±7.08) d with 80 left lower limbs and 13 right lower limbs involved. Results There were 30 patients with prior IVC filter implanted compared with 63 patients without filters (67. 7% ,63/93). Among the patients without filter, there were 93. 6% (59/63) left lower limbs. DSA was redone after thrombolysis. Seventy-seven iliacfemoral thrombosis was resolved completely and stenosis or occlusion of the iliac vein were found in 90. 9% (70/77) cases. Endovascular treatment was performed in 57 patients. There was no patients suffering from symptomatic pulmonary embolism (PE). PE was found in 3 cases with a filter and 1 case without filter was suspected of PE on pulmonary CTA after treatment. Conclusion It is not necessary to routinely place an IVC filter during CDT for treating DVT on left lower limb when the thrombosis is not involing the IVC.

The expression of heat shock proteins(HSPs) can protect against acute lung injury(ALI).However,HSPs are restrained from clinical application due to the toxicity of most of the former inductors.Glutamine,which also has the ability to induce the expression of HSPs,can protect against ALI and sepsis,and may serve as a candidate for clinical application.

Objective To investigate the in vivo effects of glutamine(Gln) on inflammatory cytokine release in murine peritoneal macrophages during sepsis. Methods Sixty Kunming mice were randomly divided into sham-operation group(Sham group,n=20),operation control group(Con group,n=20) and Gln-treatment group(Gln group,n=20).Sepsis was induced by cecal ligation and puncture in Gln group and Con group,and Gln(0.75 g/kg) or saline was immediately administered via single tail vein injection.Serum was collected and macrophages were harvested from peritoneal lavage at 6 h in these three groups.Intracellular and serum cytokines of tumor necrosis factor-?(TNF-?),interleukin(IL)-6 and IL-10 were detected by ELISA.The expression of TNF-? mRNA in macrophages was analyzed by RT-PCR,and the expression of heat shock protein(HSP) 72 in macrophages was evaluated by Western blotting. Results Gln group demonstrated significantly lower intracellular TNF-? and IL-6 levels than Con group(P0.05).The serum TNF-? level was significantly lower in Gln group than in Con group(P

Background5-Nitro-2-(3-styrene-acrylic amine) benzoic acid ( NPPB),a chloride channel inhibitor,has a promoting effect on cell apoptosis in myocardial ischemia and reperfusion of domestic rabbit.The CIC chloride channel has been found in the ocular trabecular cells.However,the effect of NPPB on the shape and function of trabecular cells is unclear. Objective This study was performed to investigate the effect of NPPB on the proliferation,cell cycle progression and apoptosis of human trabecular meshwork cells.MethodsThe immortalized human trabcular cell strain was cultured,and logarithmic-phase cells were incubated in 96-well plates at a density of 1 ×106/ml.Different concentrations of NPPB (10,50,100 μ mol/L) were added to the medium,and the MTT assay was used to assess the growth and proliferation of the cells.Flow cytometry was used to evaluate the cell cycle.Then,100 mg/L 5-FU or 100 mg/L 5-FU + 100 μmol/L NPPB was used to induce cell apoptosis,which was assessed by Annexin V-PI.The membrane potential of mitochondria was examined using rhodamine 123 (△ψm).Results After 48 hours of treatment with NPPB,the abosorbency (A value) of the cells was gradually lowered with the increasing dose of NPPB,with significant differences among the 4 groups (F =7.230,P =0.006).Compared with the 10 μmol/L NPPB group,the A values were significantly declined in the 50 and 100 μmol/L NPPB groups (t =1.610,P =0.025 ;t =12.270,P =0.001 ).Forty-eight hours after exposure to NPPB,the percentage of cells in G0/G1 phase was increased and that in the S phase was decreased.The percentages of cells in different phases of cell cycle were significantly different in comparison with their control groups (without NPPB)( P＜0.05 ).Twenty-four and 48 hours after the treatment with 100 mg/L 5-FU,the apoptosis rates of the cells were raised in the 100 mg/L 5-FU group and 100 mg/L 5-FU + 100 μmol/L NPPB group compared to the without NPPB group (t24h =2.130,P =0.023;t48h =4.810,P=0.011 ) ;while that in the 100 mg/L 5-FU+100 μmol/L NPPB group was higher than the 100 mg/L 5-FU group ( t24 h =1.980,P =0.037 ; t48 h =1.290,P =0.028 ),and the mitochondrial membrane potential was lowered ( t24h =1.580,P =0.029 ; F48 h =6.200,P =0.015 ).Conclusions NPPB suppresses the proliferation of human trabecular cells and promotes the cells to enter S phase via the G1/S check point.In addition,ClC might be involved in an anti-apoptosis mechanism through the internal mitochondrial pathway.

Objective To analyze the setup error during tumor radiotherapy by linear accelerator cone beam CT(CBCT),and to determine the presence of correlation between subcutaneous fat thickness and setup error. Methods From May 2016 to June 2017 59 patients with pelvic tumors underwent CT scan after body positioning with thermoplastic model,CBCT images were obtained before treatment and registered automatically and manually along with localizable CT images. The displacement data at X(left-right), Y(head-foot) and Z(front-back) directions as well as pelvic subcutaneous fat thickness were recorded,and SPSS 18.0 was used for data statistics.Results The 59 patients had the mean setup errors at X,Y and Z directions being(1.07±2.18),(-0.07±5.00)and(1.15±2.38)mm respectively,and the subcutaneous fat thickness ranging from 0.3 to 2.9 cm with a mean value of 1.6 cm.Spearman analysis showed the correlations between the absolute value of the errors and the subcutaneous fat thickness being 0.15, 0.11 and 0.20 respectively at X, Y and Z directions, and there were no significant differences between the correlations(P>0.05).Conclusion The setup method verifies its feasibility by restraining its errors at X, Y and Z directions between -5 and +5 mm, and there is a weak correlation between the setup error and subcutaneous fat thickness.[Chinese Medical Equipment Journal,2018,39(5):64-67]

OBJECTIVETo build a type 2 diabetes mellitus rat model with cardiac ischemia.

METHODSMale Wistar rats were fed high sucrose and high fat diet for four weeks and then injected with streptozoticin (STZ) (40 mg/kg .i.p.). The levels of fasting blood glucose and serum insulin were monitored every week. The body weights of rats were also measured every week. The blood levels of creatine kinase and lactate dehydrogenase (LDH) were measured following the electrocardiograph used BL-410 biological experiment system.

RESULTSThe serum insulin levels of diabetic rats were 4.05 ng/ml after four weeks high sucrose and high fat diet. The fasting blood glucose levels of diabetic rats were 17.9 mmol/L after injection. Compared with normal group, there was obvious change of S-T segment in the electrocardiograph of diabetic group at the fourteenth week. The levels of creatine kinase and lactate dehydrogenase in diabetic group significantly increased in comparison with those in normal group.

CONCLUSIONThe cardiac ischemia of diabetic rats model is suitable for investigating cardiac disease of diabetes mellitus.

Animals ; Creatine Kinase ; blood ; Diabetes Mellitus, Experimental ; physiopathology ; Diabetes Mellitus, Type 2 ; chemically induced ; physiopathology ; Diet, High-Fat ; adverse effects ; Dietary Sucrose ; adverse effects ; Disease Models, Animal ; L-Lactate Dehydrogenase ; blood ; Male ; Myocardial Ischemia ; physiopathology ; Rats ; Rats, Wistar ; Streptozocin

The aim of this study is to develop a rapid and accurately species typing method for Brucella isolates by using High Resolution Melting (HRM ) analysis .Six pairs of primers were used according to the reference for the sequence of pur‐pose gene .Nineteen biotypes of six species Brucella standard strains were identified by PCR‐HRM analysis and this analysis was used to detect the 35 clinical isolates .Results showed Brucella amplified specific melting curves were different from con‐trasted strains with primer Bspp .The six species Brucella standard strains have own characteristic curve shape from each oth‐ers by PCR‐HRM analysis with five pairs of primers .Thirty‐five clinical isolates of Brucella have entirely consistent with PCR‐HRM curve shape with Brucella melitensis standard strains .So ,PCR‐HRM analysis methods can accurately identify Brucella strains ,especially clinical isolated Brucella melitensis ,and may be used in clinical microbiology laboratories .

Objective To investigate the association between polymorphisms of thyroid-stimulating hormone receptor(TSHR)gene intron 1(rs179247, rs12101261)and Graves′ disease(GD)in the China Han population from Xuzhou city, Jiangsu Province. Methods Total 1 066 GD patients and 1 107 control subjects were recruited for genotyping by Taqman probe technique on Fluidigm EP1 platform. Meanwhile, serum concentrations of thyroid hormone and TSH receptor antibodies(TRAb)were determined. Results The rs179247_A, rs12101261_T were significantly associated with GD risk(OR=1.35, 95%CI 1.19-1.54, P=5.92×10-6; OR=1.32, 95%CI 1.16-1.50, P=2.22×10-5). Logistic regression identified that rs179247 was an independent susceptibility locus of GD. Serum TRAb concentration showed a significant difference(P=0.015)among rs179247_AA, AG, and GG genotypes. Conclusion rs179247 and rs12101261 in TSHR intron 1 are both associated with GD, and rs179247 may contribute risk to GD independently. The polymorphism is associated with TRAb, but not with serum concentration of thyroid hormones, age of onset, diffused thyroid goiter, ophthalmic signs, and relapse.

Objective To investigate the effect of 17β-estradiol on ketamine-induced long-term cognitive deficits in neonatal rats.Methods 80 SD male rats aged 7 days were randomly divided into group C,V,E,K and K+E,and 16 per group.Group C was intraperitoneally injected with same volume of saline for three consecutive days,Group V was subcutaneously injected with same volume of sesame oil for three consecutive days,Group E was subcutaneously injected with 600 μg · kg-1 17β-estradiol for three consecutive days,group K was intraperitoneally injected with 75 mg · kg-1 ketamine for three consecutive days,group K+E was intraperitoneally injected with 75 mg · kg-1 ketamine in combination with 600 μg · kg-1 17β-estradiol injected subcutaneously for three consecutive days.At 2 months of age,learning and memory abilities were tested with the Mortis water maze.After Morris water naze test,ten rats from each group were decapitated and the prefrontal cortex (PFC) was isolated to detect acetylcholine esterase(AchE) activity with ELISA assay and to measure acetylcholine(Ach) level by hydroxylammonium chloride method.Results The escape latency ((40.26±2.36) s,(30.25±2.20) s,(21.55±2.42) s) and path length((1019.35±58.13) cm,(811.16±27.58) cm,(598.34±34.74) cm) of group K were more than those of group C on the third,fourth aud fifth training days (all P＜0.05),while escape latency ((29.46±2.20) s,(24.86± 2.14) s,(17.20±1.91) s) and path length((913.90±41.89) cm,(729.42±31.36) cm,(487.64±18.61)cm) of group K+E were significantly lower than those of group K(all P＜0.05).On test day 6,rats were subjected to a probe trial,ratio of time spent in the target quadrant ((24.5±2.7) ％) and the number of crossings over previous platform locations (1.9±0.5) in group K were fewer than those of group C (all P＜0.05),while ratio of time spent iu the target quadrant((42.3±3.0) ％) and the number of crossings over previous platform locations(3.5±0.5) of group K+E were more than those of group K (all P＜0.05).The AchE activity((0.69±0.04) U · mg pro-1) in rats PFC of group K was significantly higher than that of group C ((0.52±0.06) U · mg pro-1) (P＜0.05).The AchE activity of group K +E ((0.58±0.12)U · mg pro-1) was lower than that of group K(P＜0.05).The Ach level ((2.59±0.34)mg · g-1) in rats PFC of group K was significantly lower than that of group C ((4.35±0.56) mg · g-1) (P＜0.05).The Ach level of group K+E((3.88±0.61) mg · g-1) was higher than that of group K(P＜0.05).Conclusions These results indicate that ketamine impairs learning and memory abilities as rat matures,while 17β-estradiol treatment improves these impairments by inhibiting AchE activity and increasing Ach level.