BACKGROUND:Tumor necrosis factor-α(TNF-α), a main cytokine inducing chondrocyte apoptosis of osteoarthritis, plays a regulatory role in the process of osteoarthritis. OBJECTIVE:To compare the rat models of chondrocyte apoptosis induced by different mass concentrations of TNF-α, thus providing theoretical basis for further study on the regulation of drugs on chondrocyte apoptosis. METHODS:Chondrocytes were isolated from the knee cartilage of 4-week-old Sprague-Dawley rats of clean grade by mechanical l col agenase digestion and were then incubated with different mass concentrations of TNF-αto induce apoptosis. The morphology of chondrocytes was observed under inverted phase contrast microscope, cel s were identified using immunohistochemical staining of type II col agen, as wel as the cel viability and apoptosis were detected by MTT and DAPI, respectively. RESULTS AND CONCLUSION:(1) In vitro, the cytoplasm of chondrocytes was stained brown-yel ow by using immunohistochemical staining of type II col agen. (2) At 48 hours, the apoptosis rate of chondrocytes induced by 10, 20 and 30μg/L TNF-αwas significantly higher than that of the 0μg/L TNF-α(P<0.01), and the apoptosis rate of chondrocytes induced by 40μg/L TNF-αwas significantly higher than that of the 10μg/L TNF-α(P<0.01). (3) The viability of chondrocytes induced by 10, 20 and 40μg/L TNF-αwas significantly lower than that of the 0μg/L TNF-α(P<0.01). In detail, the viability of chondrocytes induced by 20μg/L TNF-αwas lower than that of the 10μg/L TNF-α(P<0.05);the viability of chondrocytes induced by 40μg/L TNF-αwas significantly lower than that of the 10 and 20μg/L TNF-α(P<0.01, P<0.05). (4) These results suggest that 20μg/L TNF-αcan successful y induce the chondrocyte apoptosis model.

Objective To investigate the effects of protein active composition of Scorpio on apoptosis of L1210 tumor cells for the purpose of establishing the quality evaluation method of biological effect of Scorpio. Methods L1210 cells were examined by trypan blue exclusion. The proliferation of cells was determined by improved MTT assay. Flow cytometry was performed to analyze cell apoptosis with propidium iodide (PI). Results When the concentration of protein active composition of Scorpio exceeded or equaled 37 mg/ml, the coefficient correlation of the growth inhibiting curve of L1210 cells was 0.9357, and IC50 was 175 mg/ml. The excellence time was 0 to 48 hours. When the concentration of protein active composition of Scorpio exceeded or equaled 9.25 mg/ml, the apoptosis ratio of L1210 cells was raised significantly.Conclusion The protein active composition of Scorpio might promote the apoptosis and restrain the proliferation of L1210 cells. The value of anti-tumor biological effect of the protein active composition of Scorpio was 9.25 ～ 175 mg/ml. This value may be one of the indexes for quality evaluation of biological effect of Scorpio.

OBJECTIVE:To establish the method for simultaneous determination of 6 components in Lonicera japonica,and to study the content changes of them before and after before and after carbonized. METHODS:UPLC method was adopted. The deter-mination was performed on Agilent Eclipse Plus C18 RRHD column with mobile phase consisting of 0.1% phosphoric acid solu-tion-acetonitrile(gradient elution)at the flow rate of 0.2 mL /min. The detection wavelength was set at 350 nm,and column tem-perature was 25 ℃. The sample size was 1 μL. RESULTS:The linear ranges of chlorogenic acid,rutin,galuteolin,isochlorogenic acid A,isochlorogenic acid B and isochlorogenic acid C were 21.2-424 μg(r=0.9993),1.17-23 μg(r=0.9995),2.18-43 μg(r=0.9998),5.10-102 μg(r=0.9993),2.60-52 μg(r=0.9991),4.95-99 μg(r=0.9998),respectively. RSDs of precision,stability and repeatability tests were all lower than 2.0%. Recoveries were 97.11%-99.76%(RSD=1.20%,n=6),95.20%-99.90%(RSD=2.20%,n=6),95.71%-100.30%(RSD=2.20%,n=6),95.00%-96.98%(RSD=0.88%,n=6),96.47%-103.00%(RSD=2.40%, n=6),95.78%-103.80%(RSD=3.20%,n=6). Compared with before processing,the contents of rutin,isochlorogenic acid B and isochlorogenic acid C in L. japonica were increased along with processing,the contents of chlorogenic acid and isochlorogenic acid A were decreased significantly,while the content of galuteolin had no significant change. CONCLUSIONS:The method is sim-ple,precise,stable and repeatable,and can be used for simultaneous determination of 6 components in L. japonica. Those chemi-cal components have certain changes before and after carbonized.

Objective To investigate the effects of the leflunomide active metabolite (A771726) on the expression of phorbol-12-myristate-13-acetate (PMA) -induced CD147, matrix metallo-proteinase (MMP)-2 and MMP-9 on THP-1 cells. Methods THP-1 cells were cultured in RPMI-1640 medium supplemented with 10％ fetal bovine serum. For all experiments, THP-1 cells were cultured at an initial density of 5×105/ml. Before A771726 treatment, cells were cultured with serum-free RPMI-1640 medium for 12 h, THP-1cells were co-cultured with PMA at three different concentrations of A771726 (5, 15 , 45 μg/ml) for 24 h.The mRNA expression of CD147, MMP-2 and MMP-9 was measured by real-time PCR. CD147 expression on the cells were evaluated by flow cytometric analysis. The activity of MMP-2 and MMP-9 were evaluated by gelatin zymography. Statistical differences among the groups were tested by one-way ANOVA or KruskalWallis test. Results The expression of CD147, MMP-2 and MMP-9 were upgraded by the PMA. The expression of CD147 on THP-1 cells was inhibited significantly by A771726 in a dose-dependent pattern (P＜0.01). The mean fluorescence intensity (MFI) of CD147 in positive control group was 109.5±3.8, the MFI in A771726 (5, 15, 45 μg/ml) group were 73.3±2.5, 64.5±2.3, 40.9±2.7, respectively. The expression of MMP-2, MMP-9 mRNA and the activity of MMP-2, MMP-9 in the supernatant was inhibited significantly by A771726 (P＜0.01). The expression of CD147 mRNA was not inhibited significantly by A771726 (P＞0.05).Conclusion Leflunomide active metabolite (A771726) can inhibit the expression of PMA-induced CD147,MMP-2 and MMP-9 on THP-1 Cells.

Objective Looking for the chemical components which have positive correlations with anti-inflammatiory activity of Flos Lonicerea to set up pharmacodynamic-fingerprint. Methods To gain the fingerprints of different parts by HPLC and to detect the pharmacological activities of anti-inflammation, then the correlationship between chemical components and pharmacological activities were detected by linear regression. Results Pharmacological activities of the methanol extracted part were best of all, so the fingerprint of the methanol part could represent the pharmacodynamic-fingerprint of Flos Lonicerea. Conclusion The fingerprint established by this way is more scientific and reasonable.

Objective To investigate the occurrence and risk factors of surgical site infection(SSI)in patients with colon or rectal cancer.Methods Patients who were diagnosed with colon or rectal cancer and underwent emergency or elective surgery in a hospital between January 1,2008 and December 31,2013 were monitored prospectively.General data,operation condition,and antimicrobial use of patients were analyzed,occurrence of SSI was observed every day and followed up after operation,risk factors of SSI were analyzed by univariate and multivariate analysis.Results A total of 694 patients with colon cancer(n=380)or rectal cancer(n=314)were monitored,SSI occurred in 125 patients,including 15 incisional infection and 110 organ/space infection,incidence of SSI was 18.01%;incidence of SSI in colon cancer patients and rectal cancer patients were 17.11%(65/380)and 19.11%(60/314)respectively.Univariate analysis showed that among colon cancer patients,incidence of SSI was higher in those with co-infection of other sites during perioperative period,underlying diseases,phase Ⅰcancer,and relaxation suture(all P<0.05);among rectal cancer patients,incidence of SSI was higher in those with co-infection of other sites during perioperative period,underlying diseases,obstruction,operation time>2 hours,stoma,drainage,relaxation suture,rinsing during operation,and use of antimicrobial agents>72 hours(all P<0.05);logistic regression analysis showed that the independent risk factors for SSI in colon cancer patients were underlying disease,co-infection of other sites during perioperative period,and relaxation suture(all P<0.05);independent risk factors for SSI in rectal cancer patients were underlying disease,co-infection of other sites during perioperative period,and stoma(all P<0.05).Conclusion Prevention and control measures should be taken according to risk factors of SSI in patients undergoing colon cancer and rectal cancer surgery,especially those who with chronic underlying diseases and other site infection during perioperative period;in addition,patients with colon or rectal cancer should also pay attention to relaxation suture and stoma respectively.

OBJECTIVE:To improve the HPLC-fluorometric method in the determination of telmisartan in human plasma.METHODS:Using naproxen as internal standard,the plasma was extracted with ethyl acetate.The separation was performed on Symmetry C8 with mobile phase consisted of acetonitrile -0.01mol?L-1 potassium dihydrogen phosphate buffer solution(47∶53) at a flow rate of 1.0ml?min-1.The ?Ex was set at 305nm and the ?Em was set at 365nm,and the sample size was 20?L.RESULTS:The linear range of telmisartan was 2.5～200ng?mL-1(r=0.999 8),with the lowest detectable limit at 1ng?mL-1.The average recovery of extraction was (89.91?10.07)%.Both the intra-day RSD and inter-day RSD were less than 10%.The sample showed a good stability within 24 hours after three freeze-thaw cycles and extractions.CONCLUSI-ON:The method is simple,sensitive,accurate and reproducible,and suitable for the determination and pharmacokinetic study of telmisartan in human plasma.

Object To study the "floating sugar" mechanism in the root of Achyranthes bidentata Bl. Methods The important influent factors were analyzed by orthogonal test. Results The primary and secondary orders of influent factors were surrounding temperature, relative humidity, raw drug moisture. The best preservative condition: surrounding temperature is 25 ℃, relative humidity 60% and drugs moisture 11%. Conclusion Raw drugs can be stored safely when surrounding temperature is below 35 ℃, relative humidity below 70%, raw drug moisture between 9% and 13%.

Objective To assess differences in brain activation between active and passive movement of the right hand using blood oxygen level-dependent functional magnetic resonance imaging (BOLD-fMRI). Methods Nine healthy adult right handed volunteers were studied. fMRI was performed with active and passive finger-to-finger movement. Results Right hand active and passive movement produced significant activation in the contralateral sensorimotor cortex ( SMC ), the contralateral premotor cortex ( PMC ), bilaterally in the supplementary motor area (SMA) and in the ipsilateral cerebellum. The activated brain areas were centered on the contralateral SMC and PMC and located more forward during active movement than during passive movement. The contralateral SMC was the most strongly and the most frequently activated brain area. The contralateral posterior parietal cortex (PPC) was less relevant to the hand movements. Unlike active movement, passivemovement activated more areas in the posterior central gyrus than in the anterior central gyrus. Conclusions Both active and passive movement significantly activate the brain areas which are responsible for hand movement, but there are some differences in the locations of the cortex areas activated and in the incidence activation except in the contralateral SMC.

Objective To realize the simultaneous control of two laboratory instruments by a computer. Methods On the basis of analysing the mode of communication between computer and laboratory instrument, the author expanded the computer's COM interface through the USB / COM converter. Results The author realized a computer controlling MP280 CLIA chemiluminescence analyzer and anthos 2010 enzyme tester at the same time. Conclusion User can use computer's USB interface and USB / COM converter to achieve controlling multiple instruments and equipments at the same time, as long as the communication interface of equipment is open. This method has preferable potential application.