With the advent of post-genomic era, identification of protein-protein interaction has become another hot spot of protein research and promoted the invention, development and complement of relative techniques. Major large-scale high-throughput methods, such as two-hybrid system, bacteriophage display, mass spectrometry, protein microchips and bioinformatics have provided a global view of protein-protein interaction, and are hopeful to play an important role in proteomics. Each method may have its specific strengths and weaknesses as well as differences in coverage, to some extent, the data based on these methods can complete each other. Here, recent progress of these large-scale high-throughput methods and their applications for studying protein-protein interaction are reviewed.

Objective To construct a bait plasmid in bacterial two-hybrid system.Methods A cDNA fragment encoding for rat AMP-activated protein kinase ?2(AMPK?2) was amplified by PCR and inserted into bacterial expression vector pBT.After confirmation with restricted endonuclease digestion and sequence analysis,bacterial reporter strain XL-1 Blue MRF' was transformed with pBT-AMPK?2 plasmid and the expression of the recombinant bait fusion protein was detected.To test whether the bait fusion protein had the capability of self-activation,the XL-1 Blue MRF' cells were cotransformed with the pBT-AMPK?2 plasmid and empty pTRG vector,and screened on 3-amino-1,2,4-triazole(3-AT) Selective Screening Medium plates.Results Restriction digestion and sequence analysis revealed that the AMPK?2 code sequence was correctly inserted into pBT with a right reading frame.pBT-AMPK?2 expressed ?cI/AMPK?2 fusion protein.Colonies were obtained on no 3-AT Nonselective Screening Medium plates when XL-1 Blue MRF' cells were cotransformed with recombinant pBT-AMPK?2 and empty pTRG vector,while none grew on 3-AT plates,indicating that the recombinant plasmid pBT-AMPK?2 expressed AMPK?2/?cI fusion protein correctly,and was incapable of activation of the reporter cassette in the absence of an interaction partner.Conclusion The recombinant plasmid pBT-AMPK?2 could be used as "bait plasmid" to screen cDNA library.

OBJECTIVE:To investigate the characteristics of drug release and the factors affecting the in vitro dissolubility of diclofenac potassium double-layer tablets(DPD).METHODS:UV-spectrophotometry and rotating basket method in the pha-rmacopeia of China 2000 edition were used to determine the in vitro dissolubility of DPD and Higuchi equation was adopeted to simulate the in vitro drug release.The main parameters of dissolution were stastistically analysed.RESULTS:Dissolution parameters of DPD were as follows:T0.3=0.10h,Td=3.30h,T0.9=9.19h.Hardness of tablets did not affect the dissolution rate significantly;pH of dissolution media significantly influenced on the rate.Rotation speed had a significant effect on dissolution action only at the beginning of test.CONCLUSION:DPD has good properties of fast and sustained release.Proper media is the key of in vitro drug release test,however,the ultimate results should be based on in vivo trial.

OBJECTIVE:To explore the effects of sufentanil combined with remifentanil on the hemodynamics,stress re-sponse and analgesic effect of elderly patients with abdominal surgery by general anesthesia. METHODS:170 elderly patients with abdominal surgery by general anesthesia were randomly divided into control group and observation group,85 cases in each group. All patients received general anesthesia. Control group induced anesthesia by 4 ng/kg remifentanil and maintained by 5 ng/kg;obser-vation group induced anesthesia by 3 ng/kg sufentanil and 2 ng/kg remifentanil,maintained by 0.15 mg/(kg·h)remifentanil and 3 ng/kg remifentanil. Heart rate(HR)and mean arterial pressure(MAP)before anesthesia induction,immediately intubation,1 min after intubation,abdominal entry and 1 min after extubation,norepinephrine and epinephrine levels before anesthesia induction,1 min after intubation,1 min after extubation,6 and 12 h after surgery in 2 groups were observed,restlessness and alertness/seda-tion scores after extubation,6,12,24 h postoperative visual analogue scede (VAS) score and the incidence of adverse reactions were compared. RESULTS:HR and MAP levels in 2 groups immediately intubation,1 min after intubation,abdominal entry and 1 min after extubation significantly changed,HR and MAP levels in observation group 1 min after intubation,abdominal entry and 1 min after extubation were significantly lower than control group,the differences were statistically significant(P<0.05);nor-epinephrine and epinephrine levels in 2 groups 1 min after intubation,1 min after extubation,6 and 12 h after surgery significant-ly changed,and observation group was lower than control group,the differences were statistically significant(P<0.05);restless-ness and alertness/sedation scores after extubation,6,12,24 h VAS score in observation group were significantly lower than con-trol group,the differences were statistically significant(P<0.05). There was no significant difference in the incidence of adverse reactions between 2 groups (P>0.05). CONCLUSIONS:Sufentanil combined with remifentanil can effectively stabilize hemody-namics of elderly patients with abdominal surgery,reduce the stress response levels and improve the postoperative analgesic ef-fect,with good safety.

Objective To explore the application of multilevel growth model in community intervention research. Methods The data a-bout blood pressure collected through questionnaire at baseline and 6 months, 12 months after intervention were analysed through multilevel growth model to evaluate the effectiveness of hypertension self-management. Results The blood pressures between the two groups were statistical significance at baseline,and the changing trends also were different. Age could affect diastolic blood pressure (DBP) changing after controlling the other factors. Conclusion Multilevel growth model can analyze the longitudinal data acquired from intervention research flexibly,and the result is more reliable.

Objective: To investigate the protective effects of proanthocyanidin on radiation-induced injury of AHH-1 and HIEC cells, and to explore the possible molecular mechanism. Methods: CCK-8 assay was applied to examine the survival of AHH-1, HIEC cells after γ-ray irradiation with or without proanthocyanidin pretreatment. Annexin-V/PI staining and flow cytometry analysis were used to evaluate the apoptosis and cell cycle of AHH-1 and HIEC cells. The expression of Bcl-2 protein was analyzed by Western blotting assay. Results: Pretreatment with proanthocyanidin significantly increased the cell survival rate after radiation(P<0.01). The apoptosis rates of cells in the proanthocyanidin+radiation group were (11.78% at 4 Gy[AHH-1] and 5.32% at 8 Gy[HIEC], respectively) greatly lower than those of the single radiation group (26.38% at 4 Gy[AHH-1] and 12.45% at 8 Gy[HIEC], respectively, P<0.01). Moreover, the inhibition of Bcl-2 protein expression in AHH-1, HIEC cells was attenuated in the proanthocyanidin + radiation group. Conclusion: Proanthocyanidin has satisfactory protective effect on radiation-induced cellular injury; the mechanism may be related to the attenuation of Bcl-2 protein inhibition.

Objective: To screen for the differentially expressed genes during irradiation-induced malignant transformation of human bronchus epithelium cells (BEAS-2B). Methods: Suppression subtraction hybridization (SSH) was used to construct a subtracted cDNA library of differentially expressed genes during irradiation-induced malignant transformation of BEAS-2B cells. Then the subtracted library was screened by PCR and the differential fragments were sequenced and analyzed with BLAST. Fluorescent real-time quantitative PCR was used to investigate some of the differentially expressed genes. The new EST was registered in GenBank. Results: Then 40 clones were chosen to be sequenced from the library of increased expression and decreased expression respectively according to the length of insertion element. Totally 73 sequences were obtained from the 80 sequenced clones. Forty-one sequences were decreased in the transformed cells; BLAST analysis indicated that there were 6 known sequences, 20 unknown sequences, 7 void sequences and 8 repeated sequences. Thirty-two sequences were increased in the transformed cells; Blast analysis indicated that there were 14 known sequences, 9 unknown sequences, and 9 repeated sequences. Fluorescent quantitative PCR revealed that, compared with control group, the expression of MY06, HACE1, ZNF143, and HNRPH1 were significantly increased in the radiation transforming group, with their mRNAs increased by 3.49, 29.38, 12.99 and 5.00 folds, respectively. Compared with control group, the expression of PCBP2, RPL15, and TCERG1 in the radiation transforming group was significantly decreased, with their mRNAs decreased by 1.89,48.77 and 11.95 folds, respectively. The 29 unknown sequences were registered in the GenBank (ID: EB643220-EB643248). Conclusion: The cDNA library has been successfully established for malignant transformation cellular model by suppression subtractive hybridization; the library includes a number of unknown genes. The increased gene ZNF143 is associated with cell proliferation and cell division. TCERG1, as an assistant transcription activation factor, plays an important role in the mRNA transcription and later modification. PCBP2, a Polyc connection protein, plays a modulating role in protein translation. These genes have not been reported in the radiation carcinogenicity.

Objective: To explore the relationship between the mutations in the reverse transcriptase (RT) domain of hepatitis B virus polymerase (HBV-P) gene and the occurrence and development of hepatocellular carcinoma (HCC). Methods: In this case-control study, the serum samples from 202 HCC patients and 202 chronic hepatitis B virus (CHB) carriers matching for age and gender were collected. The sequence of the RT domain of HBV-P gene was determined by direct sequencing following PCR amplification. The relationship between the mutations in the RT domain of HBV-P gene and the occurrence and development of HCC was analyzed. Results: The T895A, A904T and C955T mutations in the RT domain of HBV-P gene were all significantly associated with HCC compared to non-HCC controls (P = 0.034, 47.5% vs 37.1%; P = 0.011, 5.4% vs 1.0%; P = 0.030, 5.4% vs 1.5%). Furthermore, the Logistic multivariate analysis showed that T895A was an independent risk factor for HCC [odds ratio (OR) = 2.230, 95% confidence interval (CI): 1.230-4.044, P = 0.008]. A904T or C955T increased the risk of HCC occurrence either (OR = 6.523, 95% CI: 0.838-50.733; OR = 2.904, 95% CI: 0.599-14.093), but it did not reach statistical significance. The mutation rate of combined mutation occurrence either in A904 or C955T was 9.4% in HCC group and 2.5% in non-HCC controls (P = 0.003). The adjusted OR was 4.145 (95% CI: 1.170-14.681), demonstrating its significant association with HCC (P = 0.028). Conclusion: The mutations in the RT domain of HBV-P gene are associated with the tumorigenesis of HCC. Copyright © 2014 by TUMOR.

Objective To compare the lipoprotein(a) (Lp-a) and lipid metabolism level in rheumatoid arthritis(RA) patients and control group (healthy subjects),and evaluate the correlation between Lp-a levels and systemic inflammation in patients with RA.Methods Blood samples from 30 RA patients with positive rheumatoid factor and 30 healthy controls were collected,with same distribution of gender and age in both groups.Lipid metabolism level such as Lp-a,triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol (HDL-C),low-density lipoprotein cholesterol (LDL-C),very low-density lipoprotein cholesterol (VLDL-C) and inflammation markers such as tumor necrosis factor α (TNF-α),interleukin 6 (IL-6) and C reactive protein (CRP) were detected and analyzed statistically.Results LP-a levels were higher in RA patients than that of control group(P＜0.001),HDL-C levels were lower in RA group (P＜0.05),while TC,TG,LDL-C and VLDL-C levels showed no difference (P＞0.05).The TNF-α、IL-6 and CRP levels of RA patients were higher than that of control group(P＜0.05),and the association between higher Lp-a level and TNF-α were confirmed (r=0.753,P＜0.001).Conclusions High levels of Lp-a are often observed in RA patients,and a rising Lp-a level are associated with systematic inflammation reaction.Lp-a may be a risk factor for RA progress.

AIM: To report the use of modified four ports pars plana vitrectomy in managing a dislocated posterior chamber intraocular lens.METHODS: A young man with bilateral pseudophakia and dislocated PC-IOLs had undergone modified four ports pars plana vitrectomy in relocating a dislocated posterior chamber intraocular lens. This procedure was surgically less tedious and demanding compared with previous methods of IOL retrieval.RESULTS: No significant intra- or post-operative complication was noted. The visual recovery of the operated eye was rapid with the best corrected visual acuity at 14 months post-operation being 0.5. In contrast, the right eye without operation due to patient refusal developed total retinal detachment with hand movement visual acuity.CONCLUSION: Modified four ports pars plana vitrectomy may be a more ophthalmic surgeon friendly alternative in the retrieval of dislocated IOL.