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.

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.

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

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.

Objective To identify antimicrobial susceptibilities of community-acquired Staphylococcus aureus infections and the risk factors of severe infections.Methods Clinical data of 184 cases of community-acquired Staphylococcus aureus infections collected from 4 hospitals in Ningbo during May 2008 and May 2013 were reviewed.Microbial sensitivity test and virulence genes ( pvl and tst) detection were performed in clinical isolates, and SCCmec genotyping was performed in methicillin-resistant Staphylococcus aureus ( MRSA) strains.Binary logistic regression analysis was used to identify the risk factors for severe infections.Results Among 184 cases of community-acquired Staphylococcus aureus infections, 39 ( 21.20%) were severe cases. Staphylococcus aureus strains were highly resistant to penicillin, erythromycin and clindamycin, but more than 75% strains were sensitive to oxacillin, aminoglycosides, quinolones, rifampicin and vancomycin.Logistic regression analysis showed that advanced age (OR=1.024, 95%CI:1.005-1.043, P<0.05), malignant tumor (OR=15.288, 95%CI:1.609-145.229, P<0.05) , autoimmune diseases or long-term hormone therapy ( OR=12.102, 95%CI:2.082-70.338, P <0.01 ) were risk factors for severe community-acquired Staphylococcus aureus infections. Conclusions Strains isolated from the patients with community-acquired Staphylococcus aureus infections in Ningbo are usually sensitive to oxacillin, aminoglycosides, quinolones, rifampicin and vancomycin, which may be recommended for clinical use.Elder patients and those with malignant tumor, autoimmune diseases or long-term hormone therapy are more likely to develop severe Staphylococcus aureus infections.

Objective To investigate the clinical efficacy of laparoscopic radical gastrectomy in elderly patients with advanced gastric cancer.Methods The clinical data of 85 elderly patients with advanced gastric cancer who were admitted to the Ningbo First Hospital from January 2012 to June 2014 were retrospectively analyzed.Laparoscopic radical gastrectomy was performed on 46 patients (LRG group) and open radical gastrectomy on 39 patients (ORG group).All the patients underwent primary tumor resection for gastric cancer + D2 lymph node dissection,and the postoperative recovery plans were done according to enhanced recovery program.The volume of blood loss,number of lymph node dissected,operation time,intraoperative arterial partial pressure of carbon dioxid (PaCO2),time to anal exsufflation,indwelling time of gastric tube,time for out-off-bed activity,time for fluid diet intake,postoperative hemoglobin,duration of hospital stay and occurrence of complications in the 2 groups were analyzed.The follow-up by outpatient examination and telephone interview was carried out on patients up to August 2014.The count data were analyzed by the chi-square test and Fisher exact probability.The measurement data with normal distribution were presented as x ± s and analyzed using the t test.The t' test was used if the data were deficient.Results Surgical procedures in the 2 groups were successfully carried out and no perioperative death occurred.There was no conversion to open surgery in the LRG group.The resection margins in all the patients were negative.The operation time and number of lymph node dissection in the LRG group were (239 ±68)minutes and 27 ± 10,compared with (227 ±50)minutes and 26 ± 10 in the ORG group,with significant differences (t =0.919,0.179,P ＞ O.05).PaCO2 in the LRG group was (41 ± 5) mmHg (1 mmHg =0.133 kPa),which was significantly higher than(36 ± 5) mmHg in the ORG group (t =4.745,P ＜ 0.05).The volume of blood loss was (102 ± 44)mL in the LRG group,which was significantly less than (200 ± 120) mL in the OPG group (t' =-4.807,P ＜ 0.05).The postoperative level of hemoglobin in the LRG was (110 ± 15) g/L,which was significantly higher than (98 ± 27)g/L in the ORG group (t' =2.471,P ＜ 0.05).The time to anal exsufflation,indwelling time of gastric tube,time for out-off-bed activity,time for fluid diet intake,duration of hospital stay in the LRG group were (2.6 ± 0.7) days,(2.1 ± 0.7) days,(1.1 ± 0.3) days,(4.1 ± 0.7) days and (11 ± 4) days,which were significantly different from (4.8 ± 1.5) days,(4.0 ± 1.8) days,(4.5 ± 0.6) days,(5.9 ± 1.8) days and (18 ± 3) days in the OR G group (t' =-8.415,-6.206,-33.831,-5.879,t =9.632,P＜0.05).Eight patients in the LRG group and 15 patients in the ORG group had complications,with the incidence of complications of 17.4％ (8/46) and 38.5％ (15/39),respectively,showing a significant difference (x2 =4.748,P ＜ 0.05).Forty-four patients in the LRG group and 36 patients in the ORG gorup were followed up for 2-25 months,1 patient in the LRG group and 2 patients in the ORG group died and others had full recovery.Conclusions Laparoscopic radical gastrectomy could provide a safe and complete tumor resection for elderly patients with advanced gastric cancer compared with open radical gastrectomy,meanwhile,it can improve postoperative recovery and reduce postoperative complications in elderly patients with advanced gastric cancer.

Aim To study the role of histone acetyla-tion and its involvement in the depression-like behav-iors of rats induced by chronic unpredictable stress (CUS ). Methods Thirty male Sprague Dawley (SD ) rats were randomly divided into control group and model group.The method of solitary condition with CUS for consecutive 28 days was used to establish the rat depression model.The open-field test (OFT)and the forced swimming test (FST)were used to evaluate the depressive behaviors of rats;the real time PCR was used to detect the change of HDAC2 mRNA, and Western blot was used to determine the protein expres-sions of H3,H4,acH3 and acH4 in the prefrontal cor-tex and hippocampus of rats.Results Model group showed obvious depression-like behaviors with decrea-sing locomotive activity in OFT (P <0.01 )and in-creasing immobility time in FST (P<0.01),up-regu-lating the mRNA and protein expression of HDAC2 (P<0.0 1 ),and down-regulating the protein expression of acH3 and acH4 (P<0.01)in the prefrontal cortex and hippocampus significantly,compared with control group.Conclusion The mechanism of depressive be-haviors of rats induced by CUS may be associated with down-regulating the level of histone acetylation modifi-cation.

This paper discusses application of multimedia and network technology in Laboratory Animal Science Teaching:We have set up multimedia courseware,made animal experiment video and built network course-learning system and a good teaching effect has been achieved.Then it analyzes the problems of multimedia and network technology in Laboratory Animal Science Teaching.

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.