Objective To optimize the conditions for enhancing the refolding of a novel recombinant human(rh)endostatin and test the biological activities of refolded endostatin.Methods The partial purified inclusion bodies of rh-endostatin were dissolved with 6 mol/L guanidine-HCl followed by combination of dilution and dialysis of the dissolved endostatin.The refolded endostatin was then purified by cation-exchange chromatography.The biological activities of purified rh-endostatin were assessed by endostatin-specific monoclonal antibody and chick embryo chorioallantoic membrane assay.Results A 46% refolding yield was achieved after optimizing the refolding conditions.The purified endostatin reacted with specific anti-endostatin monoclonal antibody and showed significant inhibition of angiogenesis in chick embryo ehorioallantoic membrane assay.Conclusion The method of highest refolding yield of human endostatin was developed.This optimized method significantly promotes the application of this novel human endostatin to preclinical and clinical studies.

Objective To explore an easy,safe and effective method for correction of the hypertrophy of mandibulae angle and inferior facial enlargement.Methods Based on the complexion of mandibulae angle hypertrophy,32 patients were treated through an intraoral incision with complete separation of masseter,polish of the diagonal site,camber gonion osteotomy and mandibular marginal plasty under local anaesthesia.Meanwhile,19 patients with masseter hypertrophy were injected with botulinal toxin A three months after the operation,and 2 patients with facio-buccal fullness were treated with removal of the buddal fat pad.Results 12 patients were satisfied with the results after following-up for one to six months.Conclusion This method is easy-doing,safe,reliable and accords with the principle of aesthetics.Osteotomy and grinding of the bone are convenient,which have little damage to patients with less complications and quick recovery.

Objective To investigate the influence of antiepileptic drugs (AEDs) on levels of serum homocysteine (Hcy),folate,vitamin B12 and B6 in patients with post-stroke epilepsy (PSE).Methods The serum levels of Hcy,folate,vitamin B12 and B6 of 194 PSE patients with AEDs treatment for more than 1 year (AEDs treatment group) and 40 newly diagnosed PSE patients without AEDs therapy (control group) were detected.The effects of AEDs on above indexes were analyzed.Results Compared with control group,the serum level of serum Hcy was significantly increased,and the serum levels of folate,B12 were significantly decreased in AEDs treatment group (all P<0.05).The difference of the serum levels of vitamin B6 among the groups was not significant.Compared with monothetapy subgroup,the serum levels of Hcy was significantly increased in the combination therapy subgroup (P<0.05).Compared with control group,the serum levels of Hcy were significantly increased in patients with Valproate (VPA),Carbamazepine (CBZ) and Oxcarbazepine (OXC) monotherapy,the serum levels of folate were significantly decreased in patients with VPA and CBZ monotherapy,and the serum level of B12 was significantly decreased in patients with VPA monotherapy (all P<0.05).Compared with control group,the serum levels of Hcy were significantly increased in patients with 2 kinds of AEDs combination treatment [VPA+CBZ,VPA+Levetiracetam (LEV),VPA+OXC,CBZ+LEV] or ≥3 kinds of AEDs combination treatment,the serum levels of folate was significantly decreased in patients with 2 kinds of AEDs combination treatment (VPA+LEV,VPA+OXC,CBZ+LEV) or ≥3 kinds of AEDs combination treatment,the serum levels of B12 were siginificantly decreased in patients with 2 kinds of AEDs combination treatment (VPA+CBZ,VPA+OXC,CBZ+LEV) or ≥3 kinds of AEDs combination treatment (all P<0.05).The incidence of hyperhomocysteinemia (HHcy) in AEDs treatment group (36.6%) was significantly higher than that in control group (20.0%) (χ2=4.085,P=0.043).And the difference of HHcy incidence between the combination therapy subgroup (47.6%) and the control group was statistical significant (χ2=6.950,P=0.008).The difference of HHcy incidence between the monotherapy subgroup (33.6%) and the control group was not significant.The HHcy incidence of patients with VPA and CBZ monotherapy (40.5%;43.8%) were significantly higer than those in the control group (χ2=3.871,P=0.049;χ2=4.726,P=0.030).The differences of HHcy incidence between patients with OXC,LEV monotherapy (29.2%;22.9%) and the control group were not significant.Conclusions AEDs therapy has little influence on the serum levels of vitamin B6,while has great influence on the serum levels of Hcy,folate and vitamin B12.Combination treatment of AEDs and monotherapy of VPA,CBZ may increase the incidence of HHcy in PSE patients.

Objective To investigate the clinical diagnostic and differential diagnostic values of antisense non-coding RNA in the INK4 locus (ANRIL) and tumor suppressors (p14ARF, p15INK4b and p16INK4a) mRNA expression levels in peripheral blood lymphocytes of patients with cirrhosis and hepatocellular carcinoma. Methods The patients with hepatocellular carcinoma and cirrhosis admitted in Shantou Central Hospital from October 2013 to April 2014 were selected. The real-time quantitative reverse transcription-polymerase chain reaction (RT-qPCR) was used to detect ANRIL, p14ARF, p15INK4b and p16INK4a mRNA expression levels of peripheral blood lymphocytes. The subjects having taken physical health examinations in outpatient clinics were assigned in the healthy control group. Results During the study period, 19 cases of hepatocellular carcinoma, 24 cases of cirrhosis, and 31 healthy controls were finally enrolled. In the hepatocellular carcinoma group, the mRNA expression level of ANRIL was significantly higher than that of the healthy control group (?Ct:13.07±0.62 vs. 12.45±0.84, P<0.01), while p15INK4b mRNA expression level was obviously lower than that of the healthy control group (13.24±0.98 vs. 13.99±0.99, P<0.05). But there were no significant differences in the mRNA expression levels of ANRIL (13.07±0.65 vs. 12.71±0.76) and p15INK4b (13.24±0.98 vs. 13.55±1.08) between the groups of hepatocellular carcinoma and cirrhosis (both P>0.05). There were also no statistically significant differences in p14ARF and p16INK4a mRNA expressions among the three groups (all P>0.05). Conclusion The elevation of ANRIL and descent of p15INK4b mRNA expression levels in peripheral blood lymphocytes in patients with liver lesion can be used as the reference indicators for the early diagnosis of hepatocellular carcinoma and to predict their prognoses.

Objective:To observe recovery of the pelvic floor muscle by using electrical stimulation biofeedback system in first-time mothers after natural birth.Methods: One hundred an seventy-two cases of primipara by transvaginal full-term natural birth, less than 35 years old, myodynamiaⅢ or less level were choosed for research object between January 2013 and January 2014 in our hospital. According to admission order, an odd number into the control group, even into the treatment group, 93 cases in each group. Kegel exercise method was used to pelvic floor muscle rehabilitation in control group, on the basis of the treatment in control group, recovery of postpartum pelvic floor muscle was conducted by using electrical stimulation biofeedback apparatus in treatment group. Strength of maternal pelvic floor muscle after recovery was tested with neuromuscular stimulation therapy apparatus and to investigate rate of urinary incontinence and sexual satisfaction in two groups.Results:①The cases ofⅠ,Ⅱ class A and BⅡ pelvic floor muscle fiber restoring toⅣ andⅤ increased significantly after electrical stimulation biofeedback system treatment in treatment group, compared with control group, the difference was statistically significant.The pelvic floor muscle strength increased significantly after treatment in treatment group (P<0.05).②Urinary incontinence rate after treatment for 2 months and 6 months in treatment group was lower significantly than that in control group, sexual life satisfaction was higher significantly than that in control group, differences between groups were statistically significant (P<0.05).Conclusion: Application of electrical stimulation biofeedback system in the treatment of female pelvic floor muscle dysfunction can enhance the enthusiasm of the patients in treatment and curative effect is reliable, and no obvious complications occurred, is worth further promotion in clinical.

This review focus on the perspective of genetic engineering technology in biopharmaceutical field.Four general aspects of genetic engineering are further discussed in this review as following: the separation of macromolecules,PCR,gene chip,and the expression of foreign genes.The large-scale or industrialization of genetic engineering technology in biopharmaceutics is also reviewed.

OBJECTIVE:To evaluate the economics of penehyclidine hydrochloride and atropine in the treatment of acute organophosphorus poisoning (AOPP).METHODS:The information of 118 AOPP patients were collected and divided into group A (59 cases) and B (59 cases) according to therapy plan.There were 22 cases of mild poisoning,20 cases of moderate poisoning and 17 cases of severe poisoning in group A.There were 21 mild cases,21 moderate cases and 17 severe cases in group B.Based on routine treatment,group A was given Penehyclidine hydrochloride injection intramuscularly with initial dose of 1 mg (mild),2 mg (moderate) and 4 mg (severe).Group B was given Atropine sulfate injection intravenously,with initial dose of 2 mg (mild),5 mg (moderate) and 10 mg (severe).Both received maintenance treatment according to patients condition and stopped taking medicine after symptoms disappeared.Clinical efficacies,the time of acetylcholinesterase recovery and ADR were observed in 2 groups.The economics of therapy plans for mild,moderate and severe poisoning were evaluated in 2 groups by cost-effectiveness analysis.RESULTS:There was no statistical significance in total response rate of mild poisoning or the time of acetylcholinesterase recovery between 2 groups (P＞0.05).Total effective rates of moderate and severe poisoning in group A were significantly higher than group B,and the time of acetylcholinesterase recovery was significantly shorter than group B,with statistical significance (P＜0.05).There was no statistical significance in the incidence of ADR in mild,moderate and severe poisoning patients (P＞0.05).Cost-effectiveness ratio of penehyclidine hydrochloride was similar to that of atropine in mild poisoning patients;that of penehyclidine hydrochloride were significantly lower than that of atropine in moderate and severe patients.It was inline with the results of sensitivity analysis.CONCLUSIONS:Based on routine treatment,penehyclidine hydrochloride is similar to atropine in therapeutic efficacy of AOPP and the time of acetylcholinesterase recovery.For moderate and severe AOPP patients,penehyclidine hydrochloride is significantly better than atropine in improving therapeutic efficacy and the time of acetylcholinesterase recovery.The safety of 2 drugs are satisfactory;penehyclidine hydrochloride possesses cost-effectiveness advantage.

Objective:To purify and characterize the major antigenic components of sand swimming crab,and provide theoretical evidences for the research of standardization of allergenic vaccine.Methods:The crude extract of Linnaeus was prepared using classical method, and electrophoresis of SDS-PAGE was used to separate the proteins of each group. The allergen proteins of 26 crab were identified using Western blot. To distinguish between the primary allergen proteins and secondary allergen proteins, FPLC(Gel Chromatography and ion exchange chromatography) was used to filtrate and identify the allergen protein.Results:SDS-PAGE analysis revealed that sand swimming crab proteins were composed of at lease 9 discrete protein bands, which molecular weight ranging from 13 000 to 90 000. Major bands were of 20 900, 24 200, 27 100, 29 200, 33 700, 38 900, 48 700, 74 700, 89 100 respectively. Western blot assay indicated that the crude extract reacted with sera obtained from 26 crab allergenic subjects and contained 5 allergen bands altogether, and the bands of 74 400 and 48 700 were the major allergenic components. The positive rates of the two major allergen proteins were both 100%, indicating the allergenic components maintained the immunocompetence after yielding from chromatography.Conclusion:The 74 400 and 48 700 bands are the major allergens of sand swimming crab. The major allergen proteins can be purified by chromatography.

Interleukin-22 (IL-22) is a member of the IL-10 family, which exhibits a similar structure and immune function with IL-10. IL-22 is mainly secreted by Th22, Th1, and Th17 cells, innate lymphoid cells (ILCs), and γδT and NKT cells. IL-22 is mainly in-volved in autoimmune diseases, malignant tumors, and infective diseases. As such, studies have focused on the function of IL-22 cyto-kine in the pathogenesis of various malignant tumors. Studies revealing the expression and pathogenesis of IL-22 in malignant tumors possibly provide a new method to treat and prevent malignant tumors. In our study, the discovery, structural characteristics, and func-tions of IL-22 in malignant tumors are reviewed.

Objective To study the treatment effect by addition of granulocyte-colony-stimulating factor (G-CSF) that could reduce the level of residual disease in patients with Ph-positive chronic myeloid leukemia (CML) who appeared to have achieved a suboptimal response to imatinib (IM) alone. Methods Eleven patients with CML who had achieved≥ 35 % Ph-negativity on treatment of IM were enrolled. The initial dose of imatinib was 400 mg or 600 mg orally daily, and G-CSF, 5 μg/kg subcutaneously daily. The administration of G-CSF was postponed or interrupted in the event of leukocytosis (leukocytes ≥ 30 ×109/L) until the whitecell count fell ＜20 × 109/L. Efficacy was assessed by serial monitoring of blood levels of bcr-abl transcripts.Treatment with G-CSF was discontinued if the patient did not achieve a reduction in the transcript level of at least 0.5 log after 6 months. For patients whose bcr-abl transcript levels continued to decline but had not yet reached molecular remission, treatment was designed to continue for 1 to 6 months. Results Of 11 evaluable patients, nine had an appreciable decline in bcr-abl transcript levels(include 7 cases the reduction was greater than 1 log and 2 cases the reduction was greater than 0.5 log), 2 cases the reduction was lower than 0.5 log.In 7 cases the reduction was greater than 1 log, including five patients who did not achieved complete cytogenetic response and two patients achieved complete molecular responses. No bleeding episodes occurred.No patient discontinued therapy because of toxicity and there were no treatment-related deaths. Conclusion The addition of G-CSF should be considered safely and successfully for patients who fail to obtain optimal response to IM alone and this approach deserves further evaluation.