[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.

The professionalism view of qualified person and some reasonable proposals for the promotion of the professionalism are put forward. The necessity of the professionalization of qualified person and the effective measures of perfecting the professionalization are studied by the comparative analysis between home and abroad combined with the game model analysis. Under the current national conditions, qualified person are repositioned, and after learning the advanced experience of the EU, the registration system is strictly carried out, which can promote the professionalism of qualified person and make relevant measures in order to restrict the behavior of qualified person.

Acromegaly is mostly caused by a pituitary adenoma secreting growth hormone ( GH ) , which leads to a wide range of endocrine morbidities, with an insidious onset and slow progression.A long-term overproduction of GH results in elevated insulin-like growth factor-1 ( IGF-1 ) level, which in turn, would cause target organ damage and an increased risk for mortality and decreased quality of life.Herein, the literature regarding the progress of drug therapy for acromegaly in recent five years is reviewed.

Objective: To analyze and estimate the risks of failure modes in a pharmaceutical Co.Ltd.in Nanjing producing solid preparations by collinear production, and come up with measurements to improve the quality level accordingly.Methods: The quality risk level evaluation table for the failure modes in the various steps of collinear production was obtained through the failure mode and effects analysis (FMEA).The risk control measurements were put forward in order to control and improve different failure modes with various unacceptable risk levels.Results: The risks of solid preparations by collinear production were greatly reduced through FMEA.Conclusion: The FMEA method is an effective way to improve the quality level of collinear products.Meanwhile, there are limitations of FMEA, and other risk management methods should be combined to control the overall risk of drug production.Moreover, the failure modes at the same risk level with different properties should be analyzed and controlled accordingly.The above methods can improve the risk management level of a company and reduce the risks of cross-contamination, mistakes and air-transferring in order to enhance the efficiency of quality management system and produce safe and effective drugs.

Objective To observe the curative effect of acupuncture on insulin receptor substrate 1 ( IRS-1) , IRS-2 and glucose transporter 4 (GLUT4) in skeletal muscle of experimental rats with insulin resistance (IR) . Methods Thirty-two rats were randomly divided into normal group, model group, acupuncture normal group and acupuncture model group. The fasting plasma glucose ( FPG) level was detected with glucose oxidase method, fasting insulin and C-peptide ( C-P) were examined with enzyme linked immunosorbent assay ( ELISA) , insulin sensitivity index ( ISI) was calculated, and IRS-1 mRNA, IRS-2 mRNA and GLUT4 mRNA of skeletal muscle were observed by real-time PCR-based fluorescent assay in each group. Results Compared to the normal group, the FPG, FINS, C-P levels were increased dramatically (P<0.01), ISI, IRS-1 mRNA, IRS-2 mRNA and GLUT4 mRNA of the model group declined remarkably ( P<0.01). Compared to the model group, the FPG, FINS, C-P levels declined evidently ( P<0.05 or P<0.01) , and ISI, IRS-1 mRNA, IRS-2 mRNA and GLUT4 mRNA of the acupuncture model group were increased dramatically ( P<0.01). Conclusion The curative mechanism of acupuncture for insulin resistance is probably related to the increase of IRS-1 mRNA, IRS-2 mRNA and GLUT4 mRNA expression in skeletal muscle which contributes to the improvement of PI3K signal pathway.

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 provide reference for small interfering ribonucleic acid (SiRNA) R&D and registration in China. METHODS:The situation of SiRNA R&D at home and abroad were introduced,and problems encountered in the evaluation and approval process were summed up to put forward suggestions. RESULTS & CONCLUSIONS:SiRNA has developed slowly in Chi-na,with a gap of 5-10 years at least compared with foreign developed countries. The registration category of SiRNA is not clear;applicants lack of guidance documents;production workshop which meet requirement is in shortage;the efficiency of drug registra-tion allows of no optimism. It is recommended that pharmaceutical registration departments carry out the pilot reform for SiRNA registration,publish SiRNA registration guide,relax regulations for IND gradually,encourage applicants to participate in the inter-national multicenter clinical trials in order to improve the efficiency of SiRNA evaluation and approval efficiency.

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.