[Objective] To introduce a modified method for immunoblotting in detecting protein and to show all the details needing to be paid attention to. We take COX-2 for example. [Method] Total proteins were extracted and quantified. Denatured proteins were separated by SDS-PAGE and were transferred semi-dryly to NC membrane, then the latter was bound with specific antibody. Finally, protein expression was detected by ECL.[Result] The method was concise and easy to manipulate. The method had been successfully used to detect COX-2 protein in our lab and the pictures were reproducible. [Conclusion] Every step of the method is described in detail. It is helpful for novice to carry out immunoblotting. This established method is worthy to be recommended.

Distress thermometer (DT) is a widely used rating scale for testing distress severity in cancer patients in recent years.Recent studies have indeed showed that DT is worth being used in oncology clinic because of its acceptable validation.More and more studies have demonstrated the important role of DT on exploring distress.It may provide important basis for improving the distress in cancer patients.

Objective To investigate the serum concentrations of connective tissue growth factor (CTGF) and serum cystatin C(CysC) in diabetic nephropathy(DN) patients and its clinical significance. Methods 82 cases of diabetic nephropathy patients were divided into three groups by urinary albumin excretion rate(UAER):normal al-buminuric group(group 1),microalbuminuric group(group 2),macroalbuminuric group(group 3). Serum concentra-tions of CTGF and CysC were determined by ELISA, which was compared with 20 cases of control group. Results CTGF and CysC were markedly higher in the three groups than that in healthy controls [CTGF: (87.65±6.09)、 (105.36±7.78)、(129.05±5.12) ng/L and (68.29±5.90) ng/L;CysC: (0.99±0.10)、(1.59±0.14)、 (2.53±0.27) mg/L and (0.82±0.56) mg/L, P<0.05]. CTGF, CysC were all positively correlated with UAER (r=0.80、0.21, P<0.05), all negatively correlated with glomerular filtration rate (eGFR) (r=-0.85、-0.88, P<0.01). Conclusions The serum concentrations of CTGF and CysC may be early markers in predicting diabetic nephropathy and assessing its severity.

Objective To compare the effectiveness of the forced-air warming system and the conventional warming care practice in preventing hypothermia in patients undergoing major surgeries.Methods A total of 62 patients were enrolled in this study. Patients of both sexes,aged≥18 years,and with an ASA physical status Ⅰ-Ⅲ were included and allocated randomly into intervention group(n=30)or control group(n=32). Patients in the intervention group were provided forced-air warming,either prior to induction or during the surgery procedure depending on core temperature monitoring results. In the control group,patients were warmed with conventional warming care without any intervention. Core temperature was measured by Spot-On sensor and tympanic membrane thermometer,blood pressure,the electrocardiogram,heart rate,pulse oxygen staturation,and partial pressure of end-tidal COwere also monitored continuously in the two groups.Crystalloid,colloid,and allogeneic blood were also recorded by strict protocol. Results Intraoperative hypothermia monitored with tympanic thermometer was observed in 23 patients(71.87%)in the control group,which was significantly higher than that (0) in the intervention group(P=0.00). In addition,the incidence of hypothermia monitored by Spot-On sensor was 81.25%(n=26)in control group and 13.33%(n=4)in intervention group(P=0.00). Conclusion Compared with the conventional warming care,forced-air warming system is more effective in preventing hypothermia during major surgeries.

Objective To study the effect of follicle stimulating hormone(FSH)on the secretion of anti-mullerian hormone(AMH)in granulosa cells in patients with polycystic ovarian syndrome(PCOS).Methods From Aug.2008 to Dec.2009,33 patients with PCOS in Reproductive Medicine Research Center,Sixth Affiliated Hospital of Sun Yat-sen University were collected from 8-10 mm antral follicles and classified into the following three groups: FSH-unstimulated granulosa cells(unstimulated group,n =12),exogenous FSH-stimulated granulosa cells(exo-stimulated group,n =12)and internal FSH-stimulated granulosa cells(int-stimulated group,n =21).ELISA and real-time PCR were relatively used to measure protein and mRNA level of AMH.Luciferase reporting vector was constructed to detect the promoter activity of AMH.Results The levels of AMH secretion in PCOS granulosa cells were(11.4 ± 4.0)μg/L in unstimulated group,(7.9 ± 1.1)μg/L in exo-stimulated group and(5.6 ± 1.7)μg/L in int-stimulated group.Both the external and internal stimulation of FSH may suppress AMH secretion significantly(P ＜0.05).The mRNA level of AMH in PCOS granulosa cells were 2.5 ± 1.2 in unstimulated group,which were higher than 1.5 ± 0.5 in exo-stimulated group and 1.1 ± 0.7 in int-stimulated group significantly(P ＜0.05).The luciferase activity of AMH in PCOS granulosa cells were 11.5 ± 2.3 in unstimulated group,8.7 ± 2.4 in exo-stimulated group and 6.8 ± 2.4 in int-stimulated group.The luciferase activity of AMH in unstimulated group was significantly higher than those in exo-stimulated and int-stimulated groups(P ＜0.05).Conclusion FSH may inhibit the excessive secretion of AMH and stimulate follicle growth in PCOS granulosa cells by suppressing activity and expression of promoter.

Objective:To study the content in the new version of EU-GMP ANNEX 15 (draft) to find the changes of the new and the old version and help the domestic pharmaceutical enterprises meet the requirements of the EU-GMP and complete verification and validation activities.Methods:The new version and the old version of EU-GMP ANNEX 15 were compared,and the changes in the new ANNEX 15 were analyzed in order to understand the changes deeply.Combined with the current implementation status of verification and validation activities in domestic pharmaceutical enterprises,some suggestions on qualification and validation work were provided for domestic pharmaceutical enterprises.Results and Conclusion:The new revision of EU-GMP ANNEX 15 (draft) emphasizes qualification and validation activities during the life cycle of products,and processes,and risk management methods should be used in qualification and validation activities.

Objective To compare the clinical curative effect of partial hepatectomy combined with biliary intestinal anastomosis and T tube drainage in the treatment of intrahepatic stones.Methods 72 patients with intrahe-patic stones were selected and randomly divided into observation group and control group.The observation group received lobectomy of liver +bile duct jejunum anastomosis,the control group received lobectomy of liver +T tube drainage treatment.The operation time,intraoperative blood loss,functional exercise time,length of hospital stay, recurrence rate,clinical efficacy and complications were observed.Results The patients were followed up for 6 -12months.The operative time,blood loss,ambulation exercise time,length of hospital stay in the observation group were (158.29 ±32.53)min,(161.42 ±11.75)mL,(3.61 ±0.28)d,(5.31 ±1.17)d,which were significantly more than those in the control group[(127.57 ±26.38)min,(78.29 ±8.36)mL,(2.25 ±0.56)d,(4.86 ± 2.39)d].The recurrence rate of stone of the observation group during the postoperative follow -up was 5.56%, which was significantly lower than 16.67% of the control group (t =4.328,7.196,2.174,2.259,1.862,all P <0.05).The excellent and good rate of the observation group was 94.44%,which was higher than 83.33% of the control group,the difference was statistically significant (χ2 =1.257,P <0.05).6 cases in the observation group had complications (16.67%),including 1 case of incision infection,subphrenic infection in 2 cases,bile leakage in 2 cases,1 case of pulmonary infection.15 cases in the control group had postoperative complications (41.67%), including 3 cases of wound infection,bile leakage in 4 cases,3 cases of biliary tract bleeding,abdominal bleeding in 1 case,2 cases of pulmonary infection,subphrenic infection in 1 case,pleural effusion in 1 case,the difference between the two groups was significant (χ2 =2.291,P <0.05).Conclusion Liver resection is an effective method for treatment of intrahepatic bile duct stones,on the basis of liver resection by intestinal anastomosis in the treatment of intrahepatic bile duct stone long -term effect is better,which can effectively prevent recurrence of the intrahepatic bile duct stone and bile duct cancer,widely used in patients with indications for clinical physicians.

Objective:To discuss the regulation requirements of software based on the analysis of software-related recalls of medical devices. Methods:The data of software-related recalls of medical devices in America were collected and processed, and the type, situation, cause and trend of software-related recalls were analyzed, then the suggestions on the regulation requirements of software were proposed. Results:During the four periods from 1983 to 2011, the percentage of software-related recalls vs medical device recalls in America increased from 6%to 19%, and the year-on-year growth rate of software-related recalls rose by 47%to 164%, significantly higher than medical device recalls in the same period (from 13%to 54%). Especially, the problem of software-related recalls in radiology medical devices was the most serious. Conclusion: The regulatory situation of software is more complicated and severe, so the regulation requirements of software, including software validation, adverse event monitoring and usability design, need to be further strengthened according to the particularity of software.

OBJECTIVE: To seek approach for the protection of domestic pharmaceutical enterprises with immature creativity and to ensure drug accessibility. METHODS: An analysis was performed applying the theory of demand elasticity by comparing the attributes of health right and patent right etc based on TRIPS protocol and the related practice abroad. RESULTS & CONCLUSIONS: For a developing country, China should give top priority to the short-term urgent needs, such as to meet people's medication demands, followed by meeting the long-term demands, such as to encourage independent intellectual property rights. Hence, drug accessibility-related system in drug patent system should be evolved step by step.

OBJECTIVE: To provide references for Chinese enterprises to grasp the trend of quality management of worldwide medical products and to improve the quality management level.METHODS: FDA's CGMP Warning Letters on medical products issued by the USA during 2002～2006 were analyzed statistically,and FDA's emphasis and intent on quality management system of medical products was discussed.RESULTS & CONCLUSIONS: The contents stated in FDA Warning Letters are of great importance for Chinese enterprises to grasp the trend of quality management of worldwide medical products and to improve the quality management level.Chinese enterprises can probe into the problems and solutions of quality management of medical products by analyzing FDA's emphasis and intent on quality management system of medical products as well as field investigation.