OBJECTIVE:To study the effects of celastrol on the proliferation and apoptosis of human hepatoma HepG2 cells, and investigate its mechanism. METHODS:CCK-8 method was used to determine the cell activity 24,48,72 h after treated by 2, 5,10 μmol/L celastrol,and the proliferation inhibition rate and half inhibitory concentration(IC50)were calculated;flow cytome-try was conducted to detect the cell apoptosis rate and cycle change 24 h after treated by 2,5,10μmol/L celastrol,and the DMSO was used as negative control;rhodamine 123 staining method was used to determine the mitochondrial membrane potential 48 h af-ter treated by 2,5,10 μmol/L celastrol,and the DMSO was used as negative control;Western blot was adopted to detect the pro-apoptotic related genes Bax and B lymphoma 2(Bcl-2)protein expressions 0,12,24,36 h after treated by 5 μmol/L celastrol. RESULTS:2,5,10 μmol/L celastrol can inhibit cell proliferation,IC50 was 5.834 μmol/L. 2,5,10 μmol/L celastrol can induce apoptosis;5,10 μmol/L celastrol can block cell in G0/G1,S phases,compared with negative control group,with significant differ-ences(P<0.05 or P<0.01),and the above effects all showing certain concentration-dependent manner. 5 μmol/L celastrol can in-crease Bax protein expression and decrease Bcl-2 protein expression after cultured for 12,24,36 h,showing certain time-depen-dent manner;compared with 0 h,there was significant difference(P<0.05 or P<0.01). CONCLUSIONS:Celastrol can obvious-ly inhibit the proliferation of human hepatoma HepG2 cells and induce their apoptosis,and the mechanism may be related with strengthening mitochondrial permeability and promoting the release of apoptosis-inducing factor.

Objective To observe the effect of rosuvastatin and memantine on EPC and learning or memory ability in VaD rats .Methods Forty adult male SD rats were randomly divided into sham operation group ,VaD model group ,memantine treatment group ,rosuvastatin treatment group , and memantine+rosuvastatin treatment group (combined treatment group) (8 in each group) .A VaD model was established by permanent ligation of bilateral common carotid arteries .The ani‐mals in rosuvastatin treatment group ,memantine treatment group and combined treatment group received gastric rosuvastatin 10 mg/(kg · d) ,memantine 10 mg/(kg · d) and memantine 10 mg/(kg · d)+rosuvastatin 10 mg/(kg · d) for 4 weeks while those in model group received gastric normal saline .Five weeks after operation ,the learning or memory ability was tested by Morris water maze test ,the escape latency and percentage of target quadrant were calculated ,the circulat‐ing EPC were detected by flow cytometry ,and the MVD in hippocampaus was assyed with vWF immunostaining .Results Five weeks after operation ,the learning or memory ability was signifi‐cantly lower in model group than in sham operation group (P< 0 .01) whereas the learning or memory ability ,the percentage of circulating EPC and the MVD in hippocampus were significantly higher in 3 treatment groups than in model group (P<0 .05) .Conclusion Rosuvastatin and me‐mantine can effectively improve the learning or memory ability of VaD rats by mobilizing their cir‐culating EPC and increasing the MVD in their hippocampus .However ,the effect of memantine or rosuvastatin does not differ greatly w hen they are used alone .

Objective To investigate the effect of different investing materials on the complete denture flasking deformation.Methods The denture wax casts were invested by plaster,dental stone,and dental stone together with silicone,respectively.8 trial dentures were randomly chosen from above each group,and 4 reference points were selected in the cast.The conventional water bath polymerization system was used for the process.The distance between reference points were measured before and after thermally processing the Acrylic base.F test and Kruskal-Wallis H test were applied for statistical analysis.Results The distance between A and B (F =3.768,P =0.040) and the mean of total distance among O,A,B and C(F =4.830,P =0.019) had statistically significant differences among the three groups.The deformation variance of Group 1 (a conventional plaster) between 175μm and 267μm was maximum.The deformation values of Group2 (a dental stone) was appropriate with Group3 (a dental stone + a silicone).Smoothness (x2 =17.575,P ＜ 0.01) was statistically different among the three groups.Conclusion The flaskingprocess with the plaster had the maximum movement.The process with dental stone showed the minimal toothmovement,the process with the dental stone along with with thesilicone caused a similar movement as from the single dental stone but resulted in a smoother denture surface.

The proliferation of Patu8988 cells pre-treated with metformin for 72h was measured by CCK-8 assay.Cell cycle was analyzed by flow cytometry.The expression of related genes was detected by RT-PCR.Metformin suppressed the proliferation of Patu8988 cells in a dose-dependent manner( r=0.994,P＜0.05 ).The proportion of cells at G0/G1 stage was increased while that of G2/M stage decreased (P＜0.05 ).The expressions of MMP-3,cyclinD1,p53 were down regulated and that of Bax was up regulated.The results show that metformin may inhibit the proliferation of Patu8988 cells,via blocking the cell cycle and affecting the expression of related genes.

The study is to develop an HPLC method for simultaneous determination of rhamnazin (1), rhamnocitrin (2), rhamnetin (3), rhamnazin-3-O-beta-D-glucopyranoside (4), rhamnazin-3-O-beta-D-xylopyranosyl-(1-->4)-beta-D-glucopyranoside (5), rhamnazin-3-O-beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranoside (6), and rhamnocitrin-3-O-beta-D-glucopyranosyl-(1-->4)-beta-D-glucopyranoside (7) in Nervilia fordii. The separation was performed on a Kromasil C18 column (250 mm x 4.6 mm, 5 microm) with 0.4% phosphoric acid-acetonitrile as the mobile phase in a gradient elution at a flow rate of 1.0 mL x min(-1). The detect wavelength was set at 256 nm, and the column temperature was set at 40 degrees C. There were good linear relationships between the logarithm values of concentrations and those of the peak areas of seven flavonoids (1-7) in the range of 0.55-70.00 microg x mL(-1) (r = 0.9997), 0.86-110.00 microg x mL(-1) (r = 0.9997), 0.39-50.00 microg x mL(-1) (r = 0.999 7), 0.55-70.00 microg x mL(-1) (r = 0.999 5), 1.33-170.00 microg x mL(-1) (r = 0.9998), 1.33-170.00 microg x mL(-1) (r = 0.9998), 0.16-20.00 microg x mL(-1) (r = 0.9995), respectively. The recoveries of the seven flavonoids were between 97.19%-99.45%, the relative standard deviations (RSDs) were between 0.91%-2.69%. The established method is rapid, accurate with high repeatability, which could provide scientific evidence for the quality control of Nervilia fordii.

Objective To investigate the effect on the differentiation of bone marrow mesenchymal stem cells (BMSC) with non-contact co-culture with mechanical stimulated ligament fibroblasts. Methods A cyclic 10% uniaxia strain at 1 Hz was applied on rat pelvic ligament fibroblasts, then were co-cultured with BMSC for 3, 6 and 12 days in non-contact condition. The protein expression of collagen Ⅰ ,Ⅲ in BMSC were detected by SP method and revealed by the mean gray value. The mRNA expressions of collagens type Ⅰ and type Ⅲ in the BMSCs were measured with real-time (RT)-PCR ,and the results were indicated by the ratio between the mRNA and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) . Results (1) Protein expression; after 3 days co-culture with pelvic ligament fibroblasts, expression of collagen Ⅰ and Ⅲ in BMSC are 82. 4 ± 3. 4 and 76. 8 ± 2. 5. When compared with 80. 2 ± 2. 6 and 74. 6 ± 1. 1 in BMSC without co-culture, there was no significant difference (P > 0. 05) . After 6 days co-culture with pelvic ligament fibroblasts, the expression of collagen Ⅰ and Ⅲ of 126. 6 ±2. 2 and 118. 6 ± 1. 4 in BMSC were significantly higher than 82. 7 ±3. 0 and 76. 2 ± 1. 3 in BMSC without co-culture (P < 0. 05). Similarly, after 12 days co-culture with pelvic ligament fibroblasts, the expression of collagen Ⅰ and Ⅲ of 135. 3 ±3. 4 and 128. 7 ± 2. 6 in BMSC were significantly higher than 86. 6 ± 1. 3 and 81. 8 ± 1.4 in BMSC without co-culture (P <0.05). (2)mRNA expression:after 3 days co-culture with pelvic ligament fibroblasts , the mRNA expression of type Ⅰ and type Ⅲ collagens in BMSC are 2. 10 ±0. 20 and 1. 20 ±0. 30. When compared with mRNA expression of 2. 01 ±0. 12 and 1. 13 ±0.21 in BMSC without co-culture, no significant difference were observed (P > 0. 05). After 6 days co-culture with pelvic ligament fibroblasts , the mRNA expressions of type Ⅰ and type Ⅲ collagens mRNA were 5. 60 ±0. 21 and 2. 61 ±0. 20, which were significantly higher than 3. 70 ±0. 33 and 1. 82 ± 0. 14 in BMSC without coculture (P < 0. 05). After 12 days co-culture with pelvic ligament fibroblasts, the mRNA expressions of type Ⅰ and type Ⅲ collagens of 5. 91 ±0.31 and 2. 92 ±0. 23 were significantly higher than 4. 04 ±0. 21 and 2. 04 ±0. 13 in BMSC without co-culture (P <0. 05). Conclusion Non-contact co-culture with mechanical stretch stimulated ligament fibroblasts, it might promote synthesis of types Ⅰ and Ⅲ collagen in rat BMSCs and induced BMSC differentiated into pelvic ligament fibroblasts.

Objective To investigate thermal degradation kinetic characteristics of Cefmenoxime Hydrochloride in infusion solutions, and predict its thermal stability. Methods The HPLC was applied to determine the contents of Cefmenoxime Hydrochloride. Classical isothermal kinetic method and multivariate linear model were used to predict the expiration date of the injection. Results It was found that the thermal degradation kinetics of Cefmenoxime Hydrochloride in two infusion solutions corresponded with the first-order kinetics. The expiration dates of Cefmenoxime Hydrochloride in 0. 9%sodium chloride injection calculated by two different methods were 2. 20 days and 1. 52 days,and in 5% glucose injection were 2. 09 days and 1. 53 days,respectively. Conclusion The thermal stability of Cefmenoxime Hydrochloride in infusion solutions is poor and its expiration dates are the same calculated by two different methods.

Objective To evaluate the effects of betulinic acid preconditioning on oxidative stress response during cerebral ischemia-reperfusion (I/R) in mice.Methods Seventy-two male Kunming mice,aged 3 months,weighing 25-35 g,were randomly divided into 3 groups (n =24 each) using a random number table:sham operation group (group S),I/R group,and betulinic acid preconditioning group (group BP).Cerebral I/R was induced by middle cerebral artery occlusion in mice anesthetized with 10％ chloral hydrate 40 ml/kg.In group BP,betulinic acid 50 mg/kg was administered by intragastric gavage everyday for 7 days before ischemia,while the equal volume of solvent dimethyl sulfoxide was given in S and I/R groups.At 22 h of reperfusion,neurological function was assessed and scored.The mice were then sacrificed and brains were removed for determination of infarct size,expression of NADPH oxidase (Nox1,Nox2 and Nox4) and p22phox mRNA,activity of ROS and apoptosis rate in the infarcted zone.Results Compared with S group,neurological score,cerebral infarct size,activity of ROS and apoptosis rate in the infarcted zone were significantly increased,and the expression of Nox1,Nox2,Nox4 and p22phox mRNA was up-regulated in group I/R.Compared with group I/R,neurological score,cerebral infarct size,activity of ROS and apoptosis rate in the infarcted zone were significantly decreased,and the expression of Nox1,Nox2,Nox4 and p22phox mRNA was down-regulated in group BP.Conclusion Betulinic acid preconditioning mitigates cerebral I/R injury through inhibiting oxidative stress response in mice.

Objective To evaluate thetherapeutic effect of sodium deoxyribonucleotide injection at Zusanli point for chemotherapy-induced leukopenia in patients with lung cancer.MethodsA total of 116 chemotherapy-induced leukopenia patients with lung cancer were randomly divided into a therapy group and a control group, 60 in the treatment group and 56 in the control group. The patients in the treatment group were treated with sodium deoxyribonucleotide injection atZusnlipoint, and those in the control group were treated with vein injection of sodium deoxyribonucleotide.Results The total effective rate in the treatment group was significant higher than that in the control group (91.7%vs.76.8%;χ2=4.890,P=0.032). The effective rates on days 3,5,7 and 10 in the treatment group were also significant higher than those in the control group (on day 3: 51.7%vs. 32.1%,χ2=4.530,P=0.036; on day 5: 76.7%vs. 53.6%,χ2=6.840,P=0.018; on day 7: 85.0%vs. 67.9%,χ2=4.770,P=0.026; on day 10: 78.3%vs. 53.6%,χ2=7.960,P=0.011).Conclusion Sodium deoxyribonucleotide injection atZusanlipoint has definite efficacy for chemotherapy-induced leukopenia in patients with lung cancer.

Objective To establish a method for determining cinnamic aldehyde content in Xianggui Huazhuo capsules by gas chromatography-mass spectrometry (GC-MS). Methods The content of cinnamic aldehyde was determined by GC-MS. Separation was performed on a capillary column (30 m×0. 25 mm, 0. 25 μm) with HP-5 as the stationary phase. A programmed temperature was employed. The flow rate was 1 mL·min-1 with He as carrier gas, and split ratio was 50∶1. The injection volume was 1. 0 μL. Results The cinnamic aldehyde was well isolated from the other ingredients. A good linear relationships of cin-namic aldehyde in range of 0. 02-4. 00 mg·mL-1 was observed. The correlation coefficient was 0. 999 4. The average recovery of cinnamic aldehyde was 96. 2% , and RSD was less than 2. 11% . Conclusion The method is simple, accurate and suitable for determination of cinnamic aldehyde content.