Objective To develop and verify a whole-column image capillary isoelectric focusing(iCIEF) electrophoresis method to analyze the charge heterogeneity of recombinant human growth hormone Fc fusion protein(Fc-rhGH).Methods The iCIEF analysis method of Fc-rhGH was developed by optimizing the target protein concentration,cosolvent(urea)concentration and focusing time.The target protein was simultaneously analyzed by this method and traditional flat plate isoelectric focusing(IEF) electrophoresis,and the results were compared;The specificity,accuracy,precision,limit of quantitation(LOQ) and durability of the developed method were verified.Results The optimized method was using the mixed solution of 8 mol/L urea,0.35% methyl cellulose(MC),4% amphoteric electrolyte and 0.5% isoelectric point marker as the sample buffer,and the focusing condition was 1 500 V 1 min,3 000 V 5.5 min.IEF was not suitable for analyzing the charge heterogeneity of Fc-rhGH solution.Using the optimized iCIEF for analysis,the target protein was significantly different from the unrelated protein,and the baseline of blank reagent was stable;The recovery rate of accuracy verification was within 90%～110%,and the linear range was 0.25～0.75 mg/mL(50%～150% of the target loading volume);The RSD of each isomer pI in the repeatability verification was less than 0.3%,and the RSD of peak area percentage was less than 5%;The LOQ was 0.04 mg/ml.The sample storage time durability,amphoteric electrolyte pharmalyte 3-10 durability and MC durability of this method were good.Using this method to analyze the charge heterogeneity of Fc-rhGH physicochemical reference substance,eight charge heterogeneities of the reference substance were effectively separated,and the pI ranged from 5.9 to 6.4.Conclusion The developed iCIEF method had good specificity,accuracy,precision and durability,and was more suitable for efficient analysis of charge heterogeneity of Fc-rhGH than traditional flat plate IEF,which was of great significance for the quality control of Fc-rhGH and other Fc fusion proteins.

The analysis of the results of the renal replacement therapies,renal transplantation for 23 cases,hemodialysis for 33 cases,in 56 elderly patients with chronic renal function failure was performed.The total mortality in transplanted group with a higher early mortality is lower than that in hemodialysis group.The living quality of transplanted group was better than of the hemodialysis group.The associated diseases prior to the replacement therapies were the contributions to the death.It was believed that the age has not been served as a limitation for elderly patients receiving renal transplantation. As a replacement therapies the operation should be choosed first by elderly patients.A strictly preoperative physical examination should be done and a postoperatively special cure plan that is suited to the elderly drawn up.

Objective To investigate the clinical and pathological features of postmenopausal women with thickening endometrial. Methods Totally 187 patients with postmenopausal endometrial thickening by transvaginal B-ultrasound were divided into symptomatic group (n=104)and asymptomatic group (n=83).Their body mass index,age of menarche and menopause,history of menstruation and reproductive,application of estrogen,and hypertension and diabetes were collected.All the patients underwent hysteroscopy and biopsy procedures. Results There was no difference between two groups in body mass index [ (24.3 ± 3.2) kg/m2 vs.(23.3 ± 2.6) kg/m2,t =3.79,P =0.052]. The postmenopausal time of symptomatic patients in the group was shorter than the asymptomatic [ ( 14.6 ± 2.4) years vs.( 16.5 ± 2.9 ) years,t =4.12,P =0.039].The patients in the asymptomatic group were older than the patients in the symptomatic group [(68.1 ±4.8) years vs.(66.1± 5.5) years,F=3.89,P =0.046].The average endometrial thickness of patients in the asymptomatic group showed no difference compared with patients in the symptomatic group [(7.2 ±1.4) mm vs. (7.8 ± 2.0) mm,F=3.21,P=0.065]. There were 5 cases of endometrial adenocarcinoma,1 case of endometrial atypical hyperplasia,38 cases of endometrial polyp,25 cases of submucosal hysteromyoma,14 cases of proliferative phase endometrium in the asymptomatic group;while 8 cases of endometrial adenocarcinoma,1 case of endometrial stromal sarcoma,4 cases of endometrial serous papillary carcinoma,5 cases of endometrial atypical hyperplasia,47 cases of endometrial polyps,21 cases of submucosal hysteromyoma,2 cases of uterine empyema,8 cases of chronic inflammation of the endometrium, 8 cases of proliferative phase endometrium in the symptomatic group,respectively. Conclusions Most of endometrial thickness in postmenopausal women are benign lesions, mainly including endometrial polyps and submucous myoma, but endometrial cancer accounts for a certain percentage. Hysteroscopy is minimally invasive,safe and with specificity for the diagnosis and treatment of endometrial lesions.

Fluorescence in situ hybridization (FISH) is a kind of technique that uses fluoreseently la-beled DNA probes to assess cells for genetic alterations. UroVysion is a technique that uses FISH to detect bladder cancer in the voided urine by four fluorescently labeled DNA probes to the pericentromeric regions of chromosomes 3 ( CEP3 ) ,7 (CEPT), and 17 (CEP17) and to the 9p21 locus ( LSI 9p21 ) location of the p16 tumor suppressor gene. It is reported that sensitivity of FISH was higher than cytology for the detection of all stages and grades of bladder cancer. FDA has approved UroVysion for the detection of recurrent bladder cancer in voided urine specimens from patients with a history of bladder cancer in the year 2001 and from pa-tients with gross or microscopic hematuria but no previous history of bladder cancer in the year 2005. Fur-thermore, UroVysion can benefit the assessment of bladder cancer patients receiving BCG treatment.

Objective To investigate the clinical characteristic and prognosis of nonsteroidal antiinflammatory drugs( NSAIDs)and aspirin associated peptic ulcer bleeding. Method All patients with peptic ulcer bleeding were studied by the same researcher after admission and discharge. Results Fifty-one cases with NSAIDs and aspirin medication of the total peptic ulcer bleeding patients ( 147 cases) were included (34. 7% ). Compared with patients not associated with NSAIDs and aspirin medication, they are older [(41.2 ± 1. 9 ) years vs ( 59.4 ± 2. 2 ) years, P ＜ 0. 001], more commonly associated with hypertension ( 10. 4% vs 39. 2% ,P ＜0. 001 ), coronary heart disease ( 10. 4% vs 17.6% ,P ＜0. 001 ) ,diabetes (4. 2%vs 19. 6%, P = 0. 005 ); and had more severe anemia (7. 3% vs 23. 5%, P = 0. 008 ). Fewer patients in NSAIDs group had epigastric pain (61.5% vs 27.5%, P ＜ 0. 001 ), while there was more re-bleeding (9.4% vs 15.7%, P = 0.034). In all bleeding patients, factors associated with re-bleeding, surgical intervention and death included NSAIDs and aspirin medication, and low platelet count. In patients with NSAIDs and aspirin medication, re-bleeding was associated with previous ulcer history ( P ＜ 0. 05 ).Conclusion Peptic ulcer bleeding patients with NSAIDs and aspirin medication were more severe ill, and less likely to present with epigastric pain.

The challenge of the emergence of drug-resistant influenza strains, which is caused by wide spread utilization of direct-acting antivirals (DAAs), accelerates the research and exploration towards host targeted agents. In contrast to DAAs targeting viral replication components, host targeted agents, which regulate host factors and pathways linked to viral replication, can interfere the replication of influenza. Additionally, the innate immune system is activated by influenza during the early stage of infection, so manipulating the innate immune response may prevent the viral infection. However, the excessive inflammatory response induced at the late phase of influenza infection would lead to severe tissue injures. Thus, it is very important to explore drugs with anti-inflammatory actions to suppress these immune imbalances and tissue injures. Here we overview the current progresses about host targets related to anti-influenza drugs.

AIM: To investigate the apoptosis of a vascular smooth muscle cell line A10 caused by mild K+ depolarization. METHODS: Apoptosis was evaluated by nuclear staining, DNA fragmentation gel electrophoresis and propidium iodide-stained flow cytometry. Mitochondrial transmembrane potential (Δψm) was measured by flow cytometry. RESULTS: K+ depolarization caused dose correlated A10 cells apoptosis; nifedipine, BAPTA/AM, ryanodine inhibited the cytotoxic effect of K+ completely.The combination use of nifedipine and cyclosporin A made it clear that mitochondria was involved in the apoptosis of A10 cells,and Δψm measurement further confirmed this speculation; A10 apoptosis caused by K+ depolarization was not influenced by heparin or Zn2+,a effective capacitative calcium entry(CCE) blocker. CONCLUSION: Ca2+ entry through voltage-dependent ca channels increases intracytoplasm Ca2+, then triggers further Ca2+ release from endoplasmic reticulum via ryanodine receptor, and the microdomains of elevated intracytoplasm Ca2+ are sensed by adjacent mitochondria, which ultimately lead to cell apoptosis.

Objective To investigate the potential of AFP-targeted ultrasmall superparamagnetic particles of iron oxide (USPIO) molecular probe in specific detection of hepatocellular carcinoma (HCC) with MRI.Methods The targeted probe was synthesized by conjugating AFP antibody with modified USPIO.Two groups treated with AFP-USPIO and USPIO were set up in the study.The HepG2 cells were incubated with AFP-USPIO or USPIO (100 μg/ml) respectively with the dosage of 50 μ1,100 μl or 150 μl for 4 hours,followed by MR imaging in vitro.The signal-noise ratio (SNR) of the cells on T2-weighted image (T2WI) was measured.The rat models with orthotopic HCC were divided into two groups with 5 rats for each group at random.Pre-and post-contrast enhanced (after 1 hour) MR imaging were performed with caudal vein injection at a dosage of 20μg/ml.The contrast noise ratio (CNR) on T2WI and the difference of CNR between pre-and post-enhancement or between both groups were calculated.The relationship of SNR or CNR with the iron particles in cells or tumors was confirmed by Prussian blue iron staining.Results Cytology experiment showed the SNR in both groups was decreased with the increase of the dosage of AFP-USPIO or USPIO,indicating statistically significantdifference in SNR among three different doseage groups (P＜0.05).Prussian blue iron staining showed that the iron particles in cells were increased with the increase of AFP-USPIO dosage,and was negatively correlated with SNR (P=0.00,r=-0.926).However,the iron particles were less in cells in USPIO group.The CNRs of liver tumors in Wistar rat of pre-and post-AFP-USPIO injection were 2.05±0.88 and 0.96±0.31 respectively,indicating a significant difference (P=0.028,t=3.380).However,the CNRs in USPIO group,2.25±1.50 and 2.57±1.49,showed no statistical difference (P=0.275,t=1.263).The CNR after enhancement also had a statistical difference between both groups(P=0.042,t=3.487).Pathological results confirmed more iron particles in tumor tissues in AFP-USPIO group,whereas less in USPIO group.Conclusion AFP-USPIO molecular probes can initiatively target to the HepG2 cells and the liver cancer of rat models expressing AFP,which may help to achieve the specificity of MR imaging in the diagnosis of HCC.

Objective To investigate ultrasonographic features,clinical and histopathologic characteristics of hydrometra in the elderly women.Methods Hysteroscopy endometrial biopsy,dilation and curettage (D&C) and Tao Brush were selectively performed for the endometrial cytologic and histopathologic diagnosis according to the depth of the hydrometra investigated by transvaginal sonography,endometrial thickness and high-risk factors of carcinoma of endometrium in 72 elderly women with hydrometra.Results Among 72 elderly women with hydrometra,9 cases (12.5％) were examined by hysteroscopy,51 cases (70.8％) by D&C and 12 cases (16.7％) by endometrial cytology along with D&C.Histopathologic examination revealed that there were 7 (9.72％) cases with endometrial carcinoma,2 cases (2.8％) with atypical endometrial hyperplasia,6 cases (5.6％) with endometrial benign lesion,14 cases(19.4％)with pyometra and the other 43 cases (59.7％) with normal and simple hydrometra.Conclusions Transvaginal sonography in combination with endometrial cytology are beneficial for screening of endometrial disorders or for drainage of hydrometra in elderly women,decreasing invasive examinations such as D&C.

Objective To evaluate the efficacy and safety of cetuximab(C225) with FOLFOX4 in the treatment of metastatic colorectal cancer (mCRC). Methods 68 patients with mCRC diagnosed by pathology were randomized in two groups: cetuximab combination with FOLFOX4 chemotherapy (combined group) and FOLFOX4 chemotherapy only (control group). Results The overall response rates (RR=CR+PR)in the combined group and the control group were 57.6 % and 32.3 % with significant difference (P <0.05). Grade Ⅲ/Ⅳ toxicity were myelosuppression(27.3 % and 32.4 %, P >0.05), nausea and vomiting(12.1% and 27.7 %, P >0.05), acne-like rash (only in combined group 9.1%). Conclusion The combination of cetuximab and chemotherapy can improve treatment efficacy. Except for rash, other grade Ⅲ/Ⅳ toxicity has no difference between the two groups.