Objective] Observing Cortex Lycii Radicis' effect(Cortex Lycii) on rat insulinoma cells(INS-1) proliferation and apoptosis. To explore the mechanism of Cortex Lycii on Pancreatic Beta Cell Proliferation and apoptosis.[ Methods] After primary culture, cells were randomly divided into blank control group:control,11.1mmol·L-1 glucose,HG,30mmol·L-1 glucose, HG+Cortex Lycii(1g·L-1), HG+Cortex Lycii(2g·L-1), HG+Cortex Lycii(4g·L-1), the survival rate of cells was observed by cell counting kit-8(CCK-8);the apoptosis rate was observed by Annexin V stammg. [Results] ①Compared with HG the better effect of cell proliferation groups of Cortex Lycii(P<0.01), the best is group of Cortex Lycii(2g·L-1)(P<0.05) ②Compared with HG group the higher survival rate is group of Cortex Lycii(P<0.01), the lower apoptosis rate of Cortex Lycii(2g·L-1) compared with Cortex Lycii(1g·L-1). [Conclusion] Cortex Lycii can promote the proliferation of pancreatic beta cell, inhibit the apoptosis to protect the pancreatic beta cell. The optimal concentration is 2g·L-1.

Objective To analysis the effect of metformin on the expression of heat shock protein and VEGF in 5637 cell line of bladder cancer. Methods 5637 cell line of bladder cancer were selected, divided into blank control group, metformin 2 mM group, 5 mM group, 10 mM group and 20 mM group, 5637 cell line of bladder cancer were treated in each group.The cell proliferation inhibition rate was detected by MTT colorimetric assay, cell clone formation rate was detected by plate clone formation test, flow cytometry was used to detect cell cycle change, the expression levels of HSP-70, HSP-90α, VEGF were detected by Western blot method.Results Compared with control group, the inhibition rate of 5637 cell line of bladder cancer after 24, 48, 72 h were higher in all concentrations of metformin group(P<0.05), the higher the concentration and the longer the action time, the stronger inhibitory effect on cell proliferation.Compared with control group, the clone formation rate of 5637 cell line of bladder cancer in all concentrations of metformin group was lower, the difference was statistically significant(P<0.05).Compared with control group, the proportion of G1 phase cells were higher, the proportion of S phase cells were lower in in all concentrations of metformin group (P<0.05), the proportion of G2 phase cells in metformin 2 mM group, 5 mM group were lower(P<0.05).Compared with control group, the expression levels of HSP-70, HSP-90α, VEGF protein were lower in all concentrations of metformin group, the difference was statistically significant(P <0.05).Conclusion Metformin has obvious inhibitory effects on the expression of HSP-70, HSP-90α, VEGF in 5637 cell line of bladder cancer, can inhibit cell proliferation and cloning, promote the occurrence of G1 block, cause cell apoptosis, the effect was dose dependent.

Objective To study a new technique (lung cancer diagnossing system, LCDS) based on the computer imaging and artificial neural network for early diagnosis of lung cancer, and evaluate it's value in early cytopathological diagnosis of lung cancer. Methods The cytological smears from the specimens obtained by Percutaneous Aspiration Lung Biopsy (PALB) in 512 cases were synthetically analyzed by LCDS. Among them, 362 cases received operations. The diagnoses by LCDS were compared with postoperative histopathological diagnosis. Results In cytopathological diagnoses for the 512 specimens, LCDS can judge between cancer cells and non-cancer cells from lung lesions with its image analysis and expert system. Moreover, it can distinguish squamous carcinoma, adenocarcinoma and small cell carcinoma in cytopathological diagnosis with built-in neural network. The total coincident rate of LCDS diagnosis was 91.80% compared with the pathological diagnosis. In the 362 cases, the sensitivity of LCDS diagnosis was 94.79% (291/307), the specificity was 90.91%(50/55), and the consistent rate was 94.20%(341/362). Conclusion The diagnostic pattern of LCDS was practical and effective. It has applicable value in cytopathological diagnosis of lung cancer and may be an efficient means for early diagnosis of lung cancer.

After a description of the background and popularization of reading therapy, strategies, contents and models for the synergic innovation of reading therapy were elaborated with suggestions put forward for its implementation.

Objective To establish a UV spectrophotometry method and an HPLC method respectively for the determination of the total content of coumarin and contents of four main constituents of coumarin in Fraxini Cortex extract.Methods UV spectrophotometry was used for the determination of the content of total coumarin in Fraxini Cortex extract. The reference substance was Aesculin, and the maximum ultraviolet absorption wavelength was 334 nm. The HPLC method was used to determine the contents of Aesculin, Fraxin, Aesculetin and Fraxetin in Fraxini Cortex extract, using gradient elution with acetonitrile-phosphate solution (0.01%) as mobile phase on Agilent ZORBAX SB-C18 chromatographic column (4.6 mm × 250 mm, 5μm) at room temperature.Results For the UV method, the linear range of the mass concentration of Aesculin was 5.76-23.04μg/mL (r=0.999 9), and the average recovery was 100.6% (RSD=1.8%). For the HPLC method, the linear ranges of the mass of Aesculin, Fraxin, Aesculetin and Fraxetin were 0.055 0-3.850 0μg (r=0.9997), 0.053 9-3.773 0μg (r=0.999 8), 0.060 0-0.660 0μg (r=0.999 9), and 0.056 2-0.618 2μg (r=0.999 9), respectively, and the average recoveries were 96.97% (RSD=1.26%), 100.80% (RSD=2.22%), 99.04% (RSD=2.47%), and 98.77% (RSD=1.94%), respectively.Conclusion Both of the two methods are simple, accurate and reliable, and can be used for the quality control of total coumarin and the main constituents of coumarin in Fraxini Cortex extract.

Objective To investigate the value of bone mineral density and bone resorption markers around the mini-screw implant in the orthodontic treatment.Methods 178 orthodontic patients were prospectively collected.According to the organization condition around the mini-screw implant loaded,all patients were assigned into stable group (n =160) and loose group (n =18).The levels of interleukin-1 β (IL-1 β) and matrix metalloproteinase-9 of the liquid around the mini-screw implant were detected at 1,2 and 3 months after loading.Moreover,the bone miner al density around the mini-screw implant before and after loading were studied as well.Results Compared with the stable group,the loose groups at 1,2 and 3 months after loading got significantly higher levels of IL-1β [(35.48 ± 4.39)pg/mL vs.(29.48 ±3.92)pg/mL,t =3.348,P =0.004;(41.45 ±5.39)pg/mL vs.(26.29 ±4.12)pg/mL,t =6.493,P =0.000;(54.39 ± 12.82) pg/mL vs.(23.58 ± 3.62) pg/mL,t =11.589,P =0.000].Compared with the stable group,the loose group at 1,2 and 3 months after loading got significantly higher levels of MMP-9[(5.68 ± 3.54) ng/mL vs.(1.74 ± 0.88) ng/mL,t =8.496,P =0.000;(6.84 ± 2.82) ng/mL vs.(1.25 ± 0.62) ng/mL,t =9.835,P =0.000;(9.84 ± 4.39) ng/mL vs.(1.21 ± 0.58) ng/mL,t =12.548,P =0.000].Compared with the sta ble group,the loose group got significantly lower levels of bone mineral density of maxillary bones around the miniscrew implant before and at 3 months after loading[(620.48 ±67.82) HU vs.(694.39 ±84.58)HU,t =2.459,P =0.015;(597.39 ± 58.93) HU vs.(693.59 ± 83.29) HU,t =2.909,P =0.008].Conclusion The decrease of bone mineral density around the mini-screw implant is related to the loosening of the mini-screw implant,and IL-1 β and MMP-9 of the surrounding fluid can reflect the peripheral inflammation.

OBJECTIVE:To study the inhibitory effect of dihydrotanshinone(DTS)on human lung cancer GLC-82 cell and its mechanism. METHODS:After treated with 0(blank control),5,10,20,40,80 and 100 μg/ml DTS for 24 and 48 h,MTT as-say was used to measure the inhibition rates and IC50 of cells;cell apoptosis was detected by flow cytometry after treated with 17.85 μg/ml DTS for 12,24 and 48 h to calculate apoptotic rate;Western blot was used to detect the protein expressions of Bcl-2, Bax and Caspase-3. RESULTS:Compared with blank control group,different concentrations of DTS inhibited the proliferation of cells;24 and 48 h maximal inhibition rate were 54.48% and 64.95%,respectively;IC50 were 62.36 and 33.94 μg/ml. DTS could induce cell apoptosis in positive time dependent manner,and the range of inhibition rate was 5.6%-29.6%;Western blot showed DTS could down-regulate the expression of Bcl-2 protein and up-regulate the expression of Caspase-3 protein (P<0.01 or P<0.05). CONCLUSIONS:DTS have significant inhibitory effect on GLC-82 cells and also induce cell apoptosis,by a possible mech-anism of down-regulating the expression of Bcl-2 protein and up-regulating the expression of Caspase-3 protein.

AIM:To investigate the roles of ClC-3 chloride channels in the regulation of cell cycle and the re-lationship between ClC-3 chloride channels and the cell cycle regulators , such as cyclin D1, cyclin-dependent kinase (CDK)4, CDK6, P21 and P27 in the HeLa cells.METHODS:ClC-3 genes were silenced by the siRNA technique in the HeLa cells.The transfection efficiency of ClC-3 siRNA was detected by real-time PCR.The cell cycle distribution was ana-lyzed by the flow cytometry .The protein expression of ClC-3, P21, P27, CDK4, CDK6 and cyclin D1 was determined by Western blot .RESULTS:ClC-3 was knocked down by ClC-3 siRNA in the HeLa cells .Transfection of the cells with ClC-3 siRNA arrested the cells at G0/G1 phases, decreased the expression of cyclin D1, CDK4 and CDK6, and increased the expression of P21 and P27.CONCLUSION:ClC-3 plays an important role in the cell cycle of HeLa cells through the G 1-S transition point.ClC-3 may regulate the cell cycle progression by up-regulation of cyclin D1, CDK4 and CDK6 expression and/or by down-regulation of P21 and P27 expression.

ObjectiveTo evaluate the intermediate term efficacy of posterior stabilized (PS) total knee arthroplasty(TKA) and posterior cruciate-retaining(CR) TKA and explore the clinical related problem.MethodsThe clinical data of 84 patients (87 knees) who treated with primary TKA from May 1992 to May 2006 were analyzed retrospectively.The intermediate term efficacy was compared between PS TKA (PS group,41 cases with 43 knees) and CR TKA (CR group,43 cases with 44 knees).ResultsAll the patients were followed up for 5-10(7.6 ± 1.5 ) years,no infection,dislocation or neurovascular injury occurred in both groups.But 1 knee occurred intraoperative posterolateral femoral condyle fracture in PS group.The lateral retinacular release rate was 2.3％ ( 1/43 ) and 2.3％ (1/44) in PS group and CR group,respectively.The incidence of anterior knee pain was 4.7％(2/43 ) and 4.5％(2/44) after 6 months surgery in PS group and CR group.The incidence of anterior knee pain was 0 at the end of follow-up in both groups.There was no significant difference in the lateral retinacular release rate and the incidence of anterior knee pain between two groups (P ＞0.05).The range of motion and American knee society score(KSS) was similar and no statistical difference was found between two groups(P ＞ 0.05 ).The incidence of patellar tilt or subluxation in X-ray was 2.3％ (1/43) and 2.3％ (1/44) at the end of follow-up in PS group and CR group.There was no significant difference between two groups (P ＞ 0.05 ).At the end of follow-up,no osteolysis,X-ray radiolucent zone and prosthesis loosening were found in both groups.There was no revision owing to loosening or other reasons.ConclusionsThe intermediate term efficacy of PS TKA and CR TKA is near a tie.However,the future efficacy still need further follow-up.

Objective To evaluate the role of haeme oxygenase-1 (HO-1) in remote limb ischemic preconditioning (RLIP)-induced attenuation of lung ischemia-reperfusion (I/R) injury in rabbits.Methods Twenty-four Japanese White Rabbits,aged 4-5 months,weighing 2.0-2.5 kg,were randomly divided into 4 groups (n =6 each) using a random number table:sham operation group (S group),I/R group,RLIP group and zinc protoporphyrin (ZnPP,an inhibitor of HO-1) plus RLIP group (ZnPP + RLIP group).Lung I/R was produced by 60 min occlusion of the left lung hilum followed by 180 min of reperfusion in I/R,RLIP and ZnPP + RLIP groups.RLIP and ZnPP + RLIP groups received 3 cycles of 10 min ischemia followed by 10 min reperfusion in the bilateral hind limbs immediately before occlusion of the left lung hilum.In ZnPP + RLIP group,ZnPP 10 μmol/kg was injected intravenously 10 min prior to hind limb ischemia and the rest of the procedures were similar to those previously described in RLIP group.At the end of reperfusion,arterial blood samples were collected for blood gas analysis.The animals were then sacrificed and pulmonary specimens were obtained for microscopic examination of the pathological changes which were scored (lung injury score,LIS) and for determination of wet/dry lung weight ratio (W/D ratio),myleoperoxidase (MPO) activity,malondialdehyde (MDA) content and expression and activity of HO-1 in the lung tissues.Results Compared with group S,PaO2 was significantly decreased,and LIS,W/D ratio,MPO activity,MDA content,and HO-1 expression and activity were increased in I/R group (P ＜ 0.01).Compared with I/R group,PaO2 and HO-1 expression and activity were significantly increased,and LIS,W/D ratio,MPO activity and MDA content were decreased in RLIP group (P ＜ 0.01).Compared with RLIP group,PaO2 and HO-1 expression and activity were significantly decreased,and LIS,W/D ratio,MPO activity and MDA content were increased in ZnPP + RLIP group (P ＜ 0.01).Conclusion RLIP up-regulates HO-1 expression and enhances HO-1 activity,thus reducing lung I/R injury in rabbits.