Objective To explore the enhancement effect of caffeine in cisplatin-induced apoptosis of osteosarcoma cell line OS-U2. Methods The osteosarcoma cells were incubated with different concentrations of cisplatin (0.2, 2, 5, 10 and 20?g/L), caffeine (0.2, 2.0mmol/L) and caffeine + cisplatin for 24, 48 and 72 hours. The proliferation of OS-U2 cells was determined by MTT assay, the apoptotic levels were determined by flow cytometry (FCM), and the morphologic changes of apoptotic cells and the positive rates of apoptosis were determined by fluorescence microscopy. Results The proliferation of OS-U2 cells was inhibited, and the apoptotic level was increased when incubated with caffeine and/or cisplatin. There were dose- and time-effect relationships between the lethal effect of cisplatin on osteosarcoma cells and caffeine. Conclusion There is a remarkable enhancement effect of caffeine on cisplatin-induced apoptosis of osteosarcoma cell line OS-U2. It seems that the apoptosis-inducing activity of caffeine may enhance the lethal effect of cisplatin on osteosarcoma cells.

Objective To evaluate the comprehensive application of the CT guided transbronchial lung biopsy (CT TBLB) and CT guided percutaneous needle lung biopsy (CT_NLB) in pulmonary peripheral lesions.Methods According to the lesion location in lung field,51 patients were selected to CT TBLB and 46 patients to CT NLB.Results In the comprehensive application of the two lung biopsy methods,the comphensive biopsy success rate was 100%,pathological diagnostic positive rate 87.6% and diagnostic correct rate 97.9% (of them 100% in CT TBLB).The complications of pneumothorax and haemoptysis were decreased significantly.The positive rate and diagnostic correct rate seem higher,but there was no significant difference between the two methods (P

ObjectiveTo investigate feasibility of use of glucose injection instead of glucose powder in oral glucose tolerance test (OGTT). MethodsSixty healthy adult volunteers without history of diabetes were recruited for a standard OGTT with 75 g anhydrous glucose powder first. One week later, they were randomly divided into two groups, each of the one group (30 volunteers) orally took seven ampoules (20 ml/ampoule) and each of the other group (30 volunteer) took 7.5 ampoules of 50％ glucose injection for OGTT again, as compared to those with standard OGTT.Plasma levels of glucose and insulin were examined to evaluate whether different forms and dosage of glucose had similar results in OGTT. ResultsIn 23 volunteers with normal glucose tolerance, their plasma levels of glucose were ( 4. 8 ± 0. 4 ), ( 6. 7 ±0. 6), (5.9 ±0. 8), (5.5 ±0. 9) and (4. 8 ±0. 9) mmol/L at 0, 30, 60, 120 and 180 min after oral load with 75 g anhydrous glucose powder, respectively. These values changed to (4. 8 ± 0. 3 ), (7.5 ± 1.1 ),(6.8±1.8), (6.3 ±1.0) and (4.6 ±1.2) mmol/Lor (4.7 ±0.3), (7.2±1.3), (6.1 ±1.1),(5.6 ± 0. 9 ) and (4. 3 ± 0. 9) mmol/L after oral load with seven ampoules ( 15 volunteers) or 7. 5 ampoules of (8 volunteers ) of 50％ glucose injection, respectively.With standard OGTT, 37 cases of impaired glucose tolerance were found from 60 volunteers tested, and their plasma levels of glucose were (5. 2 ±0.6), (9. 1 ±1.4), (8.9 ±2.6), (6.7±2.0) and (4.7 ±1.0) mmol/L at 0, 30, 60, 120 and 180 min after oral load with 75 g anhydrous glucose powder, respectively; (5. 1 ± 0. 7 ), ( 8. 8 ± 1. 7 ), (9. 0 ±3.0), (7.3±2.2) and (5.1 ±1.1) mmol/L (15 volunteers) or (5.3 ±0.6), (8.8 ±1.9), (8.5 ±2. 4), (6. 6 ± 1.4) and (4. 8 ± 1.6) mmol/L (22 volunteers) at 0, 30, 60, 120 and 180 min after oral load with seven or 7.5 ampoules of 50％ glucose injection, respectively, with no statistically significant difference between varied methods.Normal serum level of insulin was found in 38 of 60 volunteers, with their logarithmic transformation of serum insulin levels of 1.5 ± 0. 3, 3.9 ± 0. 3, 3.7 ± 0. 4, 3.2 ± 0. 6 and 2.2 ±0. 8 at 0, 30, 60, 120 and 180 min, respectively after glucose load in standard OGTT, and 20 of 38 volunteers with normal serum insulin of 1.7 ± 0. 4, 3.9 ± 0.4, 3.4 ± 0. 7, 3.3 ± 0. 8 and 2. 4 ± 0. 7 at 0,30, 60, 120 and 180 min after oral load with seven ampoules of 50％ glucose injection, respectively, or 18 of 38 with normal serum insulin of 1.7 ± 0. 4, 3.9 ± 0. 4, 3.8 ± 0. 5, 3. 3 ± 0. 7 and 2. 3 ± 1.0 at 0, 30,60, 120 and 180 min after oral load with 7. 5 ampoules of 50％ glucose injection, respectively, with no statistically significant difference between varied methods. Twenty-two cases of high serum level of insulin were found from 60 volunteers with standard OGTT, with their logarithmic transformation of serum insulin of 2.2±0.6, 4.7 ±0.5, 4.9±0.7, 4.2 t 1.0 and 2. 8 ±0.9 at0, 30, 60, 120 and 180 min after oral load with 75 g anhydrous glucose powder, respectively; 10 of 22 volunteers were found with high serum insulin level of its logarithmic transformation of 2. 4 ± 0. 6, 4. 7 ± 0. 5, 4. 7 ± 0. 3, 4. 1 ± 0. 8 and 2. 8 ± 1.1 at 0,30, 60, 120 and 180 min after oral load with seven ampoules of 50％ glucose injection, respectively ; and 12 of 22 volunteers were found with high serum insulin level of its logarithmic transformation of 1.9 ± 0. 5,4. 5 ± 0. 6, 4. 6 ± 0. 6, 3. 7 ± 1.0 and 2. 4 ± 0. 9 at 0, 30, 60, 120 and 180 min after oral load with 7. 5ampoules of 50％ glucose injection, respectively; with no significant difference between varied methods.There also was no statistically significant difference in occurrence of adverse effects between these three OGTT methods. ConclusionsEither seven or 7. 5 ampoules of 50％ glucose injection can substitute 75 g anhydrous glucose powder in OGTT, with similar test results and safety.

To investigate the expression of polyadenosine diphospho-ribose polymerase 1 (PARP-1) and p16/ retinoblastoma (Rb) protein in hydroquinone (HQ)-induced TK6 cells and their regulatory mechanisms. Methods According to the 2×2 factorial design model, TK6 cells were divided into PBS-TK6 group and HQ-TK6 group based on HQ exposure, and then sub-divided into non-DOX intervention subgroup and DOX intervention subgroup based on DOX intervention, a total of four groups. The PBS-TK6 group was treated with phosphate buffer saline, and the HQ-TK6 group was treated with HQ at a final concentration of 20.0 μmol/L. The non-DOX intervention subgroup was added with 0.05% dimethyl sulfoxide; and the DOX intervention subgroup was added with PARP-1 agonist DOX at a final concentration of 0.5 μmol/L. The distribution of cell cycle was detected by flow cytometry. The protein expression of p16/Rb, cyclin D1 (cyclinD1), multifunctional protein E2 transcription factor 1 (E2F1), Rb, and p-Rb were detected by Western blot, and the level of p16 ribosylation was detected by immunofluorescence and immunoprecipitation. Results Compared with the PBS-TK6 group, the cell cycle distribution percentage in G0/G1 phase and the relative expression levels of p16 proteins were decreased in the cells of the HQ-TK6 group (all P<0.05). The cell cycle distribution percentage in S phase and the relative expression levels of cyclinD1 and p-Rb proteins were up-regulated (all P<0.05). Compared with the non-DOX intervention group, the cell cycle distribution percentage in G0/G1 and G2/M phases and the relative expression level of p16 protein increased in the DOX intervention group (all P<0.05). The percentage of cells in S phase and the relative expression levels of cyclinD1 and p-Rb proteins were down-regulated (all P< 0.05). The results of interaction effect analysis showed that compared with the non-DOX PBS-TK6 cells, the relative expression levels of Rb and E2F1 protein in the DOX PBS-TK6 cells intervention group were down-regulated (all P<0.05). The relative expression level of Rb protein in non-DOX HQ-TK6 cell group was down-regulated (P<0.05), and the relative expression of E2F1 protein was up-regulated (P<0.05). Compared with DOX PBS-TK6 cell group, the relative expression level of Rb protein in DOX HQ-TK6 cell group was down-regulated and that of E2F1 protein was up-regulated (all P<0.05). Compared with the non-DOX HQ-TK6 cell group, the relative expression level of Rb protein in the DOX HQ-TK6 cell group was up-regulated and that of E2F1 protein was down-regulated (all P<0.05). Conclusion PARP-1 participates in cell cycle regulation by regulating the p16/Rb signaling pathway in TK6 cells.

ObjectiveTo establish an ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method for detecting the bile acid expression in serum and to explore the relationship between the bile acid spectrum and the obesity.Methods After pre-treatment through solid phase extraction,serum samples were chromatographed by gradient elution on an UPLC HSS T3 column,and then mass spectrometric analysis of bile acid was performed by multiple reaction monitoring ( MRM ) mode with negative electrospray ionization (ESI).And its methodology performance, including specificity,linearity,sensitivity,imprecision and recovery,were verified according to the guideline of State Food and Drug Administration (SFDA).Furthermore,serum samples from 10 simple obesity subjects and 10 healthy controls were analyzed with this establishedmethod, and ranksum test was used for statistical analysis.Results This established UPLC-MS/MS method could simultaneously quantify 14 bile acid subgroups in serum sample,its analytical linear range was 10 - 20 000 nmol/L.For different bile acid subgroups,the limit of detection (LOD) was 0.02 - 7.90 nmol/L,limit of quantitation (LOQ) was 0.07 -44.20 nmol/L; and its within-day coefficient of variation (CV) was 0.35％ - 12.41％,between-day CV was 1.11％ - 13.04％ ; the relative recovery ratio of this method was 89.8％ - 114.6％.Some differenceswere observed in chromatogram between obesity and control subjects,and both free bile acid and conjugated bile acid concentrations in obesity subjects [ 0.49 ( 0.45 - 1.90 ),1.44 ( 0.84 - 3.72 ) μmol/L ] were lower than them in control subjects [0.98(0.53 -3.06),1.99(0.67 -2.88) μmol/L],but the difference was not significant ( Z =- 0.958,- 0.801,P ＞ 0.05 ).ConclusionsThis established UPLC-MS/MS method can efficiently differentiate and quantify 14 bile acid subgroups,and is characterized with broad analytical measurement range,good analytical sensitivity and precision.This method can be applied for the free and conjugated bile acid analysis in clinical specimens.

Objective To observe the efficacy and adverse reaction of NP and GX regimens in the treatment of the anthracycline-and-taxane-resistant advanced breast cancer.Methods Totally 75 patients with advanced breast cancer were divided into two groups,and received NP or GX regimen.NP group (n =40):NVB 25 mg/m2,day 1,day 8,iv.drip; DDP 25 mg/m2,day 1-3,iv.drip.GX group (n =35):GEM 1000 mg/m2 day 1,day 8,iv.drip; XEL 2500 mg/m2,day 1-14,bid po.Every 21 days was a cycle.The efficacy and adverse reaction were evaluated after two cycles.Results The overall response rates in the NP and GX group were 42.5 ％ (17/40) and 40.0 ％ (14/35).The median TTP of two group were 7 and 6.5 months.The MST was 15.8 and 15.0 months in the NP and GX group.The 1-and 2-year survival rates were 60.0 ％,32.5 ％ and 57.1％,31.4 ％.The increase ratio of Karnofsky were 50.0 ％ and 42.9 ％.There were not significant difference between the two groups in terms of their treatment response (P ＞ 0.05).The main adverse reactions in the two group were myelosuppression,gastrointestinal reaction and phlebitis.Hand-foot syndrome in GX was significantly higher than that in NP group,Gastrointestinal reactions in NP was significantly higher than that in GX group (P ＜ 0.05).Conclusion NP and GX regimens are effective for patients with metastatic breast cancer,their adverse reactions are tolerable,so they can be regarded as a ltermate regimens for anthracyclines and taxanes resistant patients with metastatic breast cancer.

BACKGROUND: Resin-modified glass ionomer (RMGI) cements have higher bond strength, especially can release fluoride. But there are fewer reports of the clinical application for the prevention of decalcification.OBJECTIVE: To test the benefit from using RMGI cement instead of a conventional composite resin in bracket bonding for patients with malocclusion, and observe bracket-failure rates and decalcification on enamel surfaces at pretreatment and at debonding.DESIGN: Observational and comparative trial.SETTING: The Second Hospital Affiliated to Hebei Medical University.PARTICIPANTS: Forty successive patients (358 teeth) with malocclusion admitted to the Department of Orthodontics in the Second Hospital Affiliated to Hebei Medical University, were selected for the study from July to August in 2002. All the patients (21 females and 19 males, mean age 16 years) had normal and complete anterior teeth, good oral hygiene. There were no obvious differences in bilateral teeth. Informed consents were obtained from all the subjects. The experiment was also approved by the ethical committee of the hospital. Experimental materials were RMGI adhesive (Fuji, GC, Japan, Lot 0005111) and composite resin cement (enamel adhesive of Beijing and Tianjin, Tianjin product, Lot 020402). Brackets produced from Hangzhou 3B and 37% phosphoric acid were used.METHODS: ①Bonding brackets: Subjects selected according to random procedure were divided into two groups, each with 20. GroupⅠ: The left buccal surfaces bonded with light-cure RMGI were etching for 30 seconds with 37% phosphoric acid, rinsed with water; the right buccal surfaces bonded with composite resin cement were etching for 60 seconds with 37% phosphoric acid, rinsed with water and dried; Group Ⅱ: After etching for 30 seconds with 37% phosphoric acid, the right buccal surfaces were rinsed with water and bonded brackets with light-cure RMGI. The left buccal surfaces were bonded brackets with composite resin cement after etching for 60 seconds with 37% phosphoric acid, rinsing with water and drying; Attachment of 0.036-cm NiTi wires with ligature to the brackets was conducted 10 minutes after light-curing. The information about differences in bilateral bonding materials was not told to patients. To ensure an equal bonding materials containing fluoride and minimize the error, all the patients were instructed to use toothpaste containing fluoride, a fluoride mouthwash was not prescribed. The treatment period was 9-26 months (mean 18 months).②Patients were rechecked at intervals of 4 weeks postoperatively. Each bonded tooth was checked for loose or missing brackets, and failures were recorded. A color transparency of anterior teeth area was taken using a standardized photographic technique. The enamel surface conditions were classified at a magnification of ×10. The condition of enamel surface recorded was made according to the scoring system by Geiger before treatment and at debonding.MAIN OUTCOME MEASURES: ①The number and site of bonding failures.②Enamel surface conditions at before treatment and debonding.RESULTS: Forty patients were all involved in the result analysis. Eliminating 4 teeth occurring bond failure and 4 teeth of opposite side at anterior teeth, a total of 232 teeth were evaluated.①The number and sit of bonding failures: There was no significant difference between the failure rates of RMGI adhesive and composite resin cement (P > 0.05). Significantly more premolar brackets failed than incisor brackets.②Decalcification of enamel surface: At debonding after treatment, the incidence rates of white spots in the surfaces bonded with the RMGI were significantly lower than that in the composite resin (25.9%, 38.8%, P < 0.05).CONCLUSION:The use of RMGI for brackets bonding results in a significant reduction in the incidence of white spot at debonding. Reducing etching time may obtain a similar survival rate with the routine etching time.

OBJECTIVE:To discuss the accuracy of detecting tacrolimus(FK506) concentration in whole blood by ELISA and its application in organ transplantation.METHODS:The ELISA was used to detect FK506 valley point concentrations in 5 liver transplanted recipients at different postoperative periods .Based on the results combined with clinical findings, the dosages were adjusted.The accuracy of the method and guidance action in FK506 clinical application were described.RESULTS: In 131 detections of 350 samples, the results of quality control samples were all in the control range .The detecting RSDs of high and low concentration quality control samples were 9.09% and 11.90%, respectively.The control range of FK506 concentrations in whole blood were similar to that reported in literature .CONCLUSION :ELISA is a sensitive and specific method for routine FK506 concentration monitoring in whole blood,which is suitable for application in hospital.

Objective To study the bone turnover and its related molecular mechanism in STZ-induced diabetic rats. Methods Of 30 male SD rats studied, 15 were induced diabetics by intravenous injection of streptozotocin (50 mg/kg)and fed for 8 weeks. After the sacrifice of both the diabetic and control groups, serum Ca, P, alkaline phosphatase (ALP), and osteocalcin were determined, and 24 h urinary Ca and urinary cross-linked N-telopeptide of type Ⅰ collagen (NTx)and creatinine (Cr)ratio were also determined. The left tibia was dissected for bone histomorphometry analysis. Right femur and lumbar vertebrae (L1-L4) were reserved for bone mineral density (BMD) determination. The right tibia was separated for the study of bone tissue RANKL/osteoprotegerin, Core binding factor 1 (Cbfa1) ,osterix and osteocalcin mRNA level which was performed by real-time quantitative reverse transcription polymerase chain reaction assay. Results No significant difference was found in serum Ca, P, and ALP levels between 2 groups of rats. ST-Z-induced diabetic rats were characterized by extreme hyperglycemia, marked weight loss, polyuria, and hypercalciuria. A low-turnover osteopenia was evidenced in diabetic rats by decreased BMD in both femur [(0. 099±0.013) vs (0. 139 ± 0.013 g/cm~3) , P < 0.01] and lumbar vertebrae [(0. 107±0.011)vs (0. 149±0.009) g/cm~3, P<0.01] , reduced serum osteoealcin level [a marker of formation, (3.03±0.52) vs (6. 18±0.71) ng/ml ,P<0. 01]) ,decreased urine NTx/Cr ratio [(5. 67±0.86) vs (5.23±0.98) nmol/g Cr, P<0. 05], decreased trabeeular volume and thickness, and reduced bone label surface and bone formation rate [(0. 44±0. 11) vs (0. 78±0. 14) μm/d,P<0. 01] by bone dynamic study. The RANKL/ osteoprotegerin [(0.57±0.11)vs (0.89±0.13) ,P<0.01] ,osterix [(1.93×10~(-4)±0.65×10(~-4))vs (4.19×10~(-4)± 0.71×10~(-4)) ,P<0.01] ,Cbfa1 [(26.68×10~(-4)±6.53×10~(-4))vs (37.21×10~(-4)±7.14×10~(-4)) ,P<0.01] ,and osteocalcin [(2.25×10~(-4)±1.19×10~(-4))vs (3.43×10~(-4)±1.63×10~(-4)) ,P<0.01] mRNA expressions were declined in the bone tissue of the tibia in the ST-Z-induced diabetic rats, as compared with the control. Conclusion A low-turnover osteopenia is evidenced in STZ-induced diabetic rats by significant decrease of both osteoclastic marker(RANKL/ osteoprotegerin)and osteoblastic marker (osterix ,Cbfa1 ,osteocalcin)mRNA levels in tibia.

Aim To investigate the relationship be-tween the depression-like behaviors of rats induced by chronically unpredictable stress( CUS) and the protein expression of tumor necrosis factor alpha( TNF-α) , in-dole-2 , 3-dioxygenase ( IDO ) and 3-hydroxyl amino acid oxygenase( HAAO) . Methods Thirty male Spra-gue Dawley( SD) rats were randomly divided into con-trol group and model group. CUS plus solitary condi-tion were used to establish the depression model. The open field test ( OFT ) and the force swimming test ( FST ) were used to evaluate the depression-like behaviors of rats ;Western blot was used to determine the protein expressions of TNF-α, IDO and HAAO in the prefrontal cortex and hippocampus of rats. Results Model group rats showed obvious depression-like be-haviors with increasing immobile time in FST ( P <0. 01) and decreasing locomotive activity in OFT( P<0. 01 ) , and up-regulating the protein expression of TNF-α, IDO and HAAO in the prefrontal cortex and hippocampus significantly ( P <0. 05 ) , compared with control group rats. Conclusion The depression-like behaviors of rats induced by CUS may be associated with the activation of IDO-HAAO pathway mediated by TNF-α.