The relationship between the conformation of interferon-α (IFN-α) and its anti-viral activity were analyzed by circular dichroism (CD) and flow cytometry (FCM) techniques. The recombinant human IFN-α (rIFN-α2b and rIFN-α2a) were used. CD spectra from 190 nm to 240 nm indicated that two the IFN-α showed stable secondary structure at 65 degrees C, but unstable when the temperature was above 65 degrees C, and the change was irreversible. FCM data of the anti-viral activity of IFN-α indicated that the change of its secondary structures partly weakened its anti-viral activity. The rIFN-α2b and rIFN-α2a showed the same phenomenon. These data indicated that the conformation of IFN-α is one of the factors to influence its anti-viral activity and the combination of CD and FCM is a good method to analyze the relationship between the conformation of protein drugs and their biological activities in single cell level.
Antiviral Agents ; chemistry ; Circular Dichroism ; Flow Cytometry ; Humans ; Interferon-alpha ; chemistry ; Protein Structure, Secondary ; Recombinant Proteins ; chemistry

5-Flucytosine (5-FC) could be changed to 5-fluorouracil (5-FU) by cytosine deaminase (CD), the latter is able to kill cancer cells. However, there is no efficient method to deliver the CD gene into the tumor cells, which hampers the application of the suicide gene system. In this experiment, for the first time, the NDV has been utilized as a vector to deliver the CD gene into the cancer cells, the virus can infect the cancer cells specifically, replicate and assemble, while the cytosine deaminase is expressed. Then the CD converts the prodrug 5-FC into 5-FU to achieve the purpose of inhibiting tumor. Firstly, the whole genome of E. coli JM109 was extracted, and the CD gene was obtained by cloning method. Then the CD and IRES-EGFP were ligated into the pEE12.4 expression vector to become a recombinant pEE12.4IE-CD eukaryotic expression plasmid. The human liver cancer cells were transfected with the plasmid. The cells were treated with different concentrations of 5-FC, MTT method was used to determine the killing effect of CD/5-FC system on the human liver cancer cells. The cell deaths were 18.07%, 42.98% and 62.20% respectively when the concentrations of prodrug were at 10, 20 and 30 mmol x L(-1). In 5-FC acute toxicity experiment, Kunming mice were injected with different concentrations of 5-FC at intervals of 1:0.5. The LD50 of 5-FC through iv injection was determined by improved Karber's method, the LD50 was 507 mg x kg(-1) and the 95% confidence limit was 374-695 mg x kg(-1). According to the maximum LD0 dose of the LD50, the maximum safe dose was 200 mg x kg(-1). Body weight and clinic symptoms of the experimental animals were observed. These results laid the foundation to verify the antitumor effect and safety of CD/5-FC system in animal models. The CD gene was ligated into the NDV (rClone30) carrier, then the tumor-bearing animal was established to perform the tumor inhibiting experiment. The result showed that the recombinant rClone30-CD/5-FC system has a high antitumor activity in vivo. To summarize, CD gene has been cloned and its bioactivity has been confirmed in the mammalian cells. It is the first time in this study to utilize the recombinant NDV to deliver the CD gene into the tumor cells; our result proves the rClone30-CD/5-FC system is a potential method for cancer therapy.

Objective: To summarize the methods of preventing and managing the complications in thoracoscopic lobectomy. Methods:The participants of this study included 317 patients undergoing lobectomy with video-assisted thoracoscopic surgery in the Department of Thoracic Surgery between January 2007 and December 2012. Intra-operative complications were observed, and countermeasures were summarized. Results: Complications occurred 28 times (8.8%), including bleeding in 16 cases because of accidental vascular injury (5.0%), accidental injury/break of bronchus in two cases (0.6%), vascular stump errhysis from cutting stapler in four cases (1.3%), lung stump air leakage in three cases (0.9%), lung injury in two cases (0.6%), and diaphragmatic injury in one case (0.3%). Conversion to thoracotomy was conducted in 17 cases, with a conversion rate of 5.4%. Thoracoscopic repair operation was performed in 14 cases that exhibited bleeding, with a success rate of 70% (14/20). No mortality was reported during the operation. Conclusion:Thoracoscopic lobectomy is a highly difficult method in thoracic surgeries. The procedure requires substantial attention on the timely prevention and correct management of intra-operative complications, particularly the injury and bleeding of major vessels, to reduce the rate of conversion to thoracotomy and the incidence of post-operative complications, as well as to promote the surgery in clinics.

Objective:To compare postoperative analgesic effect of flurbiprofen axetil and parecoxib sodium in patients with humeral shaft fracture in painless ward.Methods:All of 200 hospitalized humeral fractures patients were retrospectively studied in the painless ward of the Forth Medical Center of PLA General Hospital from January 2017 to September 2019 , the clinical effects of flurbiprofen axetil and parecoxib were compared.Results:Postoperative visual analogue scale (VAS) scores after 3 d of two groups of patients were significantly lower, compared with preoperative results: (4.26±0.96) scores vs. (6.09±1.38) scores, (4.04±1.19) scores vs. (6.04±1.11) scores, and the differences were statistical significantly ( P<0.01). Postoperative VAS score after 3 d of two groups had no statistical significance ( P>0.05). Two groups had different degree of adverse reactions after operation, and flurbiprofen axetil group had singnificant gastrointestinal adverse reaction: 22 patients vs. 3 patients( P<0.05). The number of patients in the parecoxib group were more than that in the flubiprofen axetil group without troubled sleep: 20 patients vs. 8 patients. Two groups of patients were satisfied with the pain care during hospitalization. Conclusions:Two analgestic drugs can obtain obvious analgesic effect in the treatment of bones surgery. The side effects and sleep disturbance in the flurbiprofen axetil group are higher than those in the parecoxib group. Good pain control can improve patients satisfaction with pain care.

This study aims to investigate the effects of fibroblast growth factor 21 (FGF-21) on learning and memory abilities and antioxidant capacity of D-galactose-induced aging mice. Kunming mice (37.1 +/- 0.62) g were randomly divided into normal control group, model group and FGF-21 high, medium and low dose groups (n = 8). Each group was injected in cervical part subcutaneously with D-galactose 180 mg x kg(-1) x d(-1) once a day for 8 weeks. At the same time, FGF-21-treated mice were administered with FGF-21 by giving subcutaneous injection in cervical part at the daily doses of 5, 2 and 1 mg x kg(-1) x d(-1). The normal control group was given with normal saline by subcutaneous injection in cervical part. At seventh week of the experiment, the learning and memory abilities of mice were determined by water maze and jumping stand tests. At the end of the experiment, the mice were sacrificed and the cells damage of hippocampus was observed by HE staining in each group. Reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) and total antioxidant capacity (T-AOC) in the brain of mice were determined. The results showed that different doses of FGF-21 could reduce the time reaching the end (P < 0.01 or P < 0.05) and the number of touching blind side (P < 0.01 or P < 0.05) in the water maze comparing with the model group. It could also prolong the latency time (P < 0.05) and decrease the number of errors (P < 0.01 or P < 0.05) in the step down test. The result of HE staining showed that FGF-21 could significantly reduce brain cell damage in the hippocampus. The ROS and MDA levels of three different doses FGF-21 treatment group reduced significantly than that of the model group [(5.58 +/- 1.07), (7.78 +/- 1.92), (9.03 +/- 1.77) vs (12.75 +/- 2.02) pmol (DCF) x min(-1) x mg(-1), P < 0.01 or P < 0.05], [(2.92 +/- 0.71), (4.21 +/- 0.81), (4.41 +/- 0.97) vs (5.62 +/- 0.63) nmol x mg(-1) (protein), P < 0.01]. Comparing with the model group, the activities of SOD, GPx, CAT and T-AOC of the three different doses FGF-21 treatment groups were also improved in a dose-dependent manner. This study demonstrates that FGF-21 can ameliorate learning and memory abilities of D-galactose induced aging mice, improve the antioxidant abilities in brain tissue and delay brain aging. This finding provides a theoretical support for clinical application of FGF-21 as a novel therapeutics for preventing aging.

To investigate the cell-killing effect and its possible mechanism of rClone30-hDR5 in combination with TRAIL on human hepatic carcinoma (HCC) cell line, first of all, recombinant plasmid pee12.4-hDR5 was introduced into HepG2 cells by liposome transfection. After five rounds of screening by flow cytometry, HepG2 cells expressing high levels of DR5 on cell surface were isolated. The cytotoxicity of TRAIL to selected cells was higher than that of TRAIL to HepG2 cells by MTT method (P < 0.01). The result suggested that the cloned hDR5 gene had biological activity. MTT assay showed that, rClone30- hDR5 in combination with TRAIL more efficiently inhibited the tumor growth of HepG2 cells compared to rClone30-hDR5 or TRAIL in vitro. The results of Annexin V-FITC/PI staining and Quantitative Real-time PCR indicated that rClone30-hDR5 in combination with TRAIL significantly increased the mRNA levels of caspase 3 and caspase 8, and induced the apoptosis of tumor cells. HepG2 cells were infected with rClone30-hDR5 or rClone30 at MOI of 1. The expression of hDR5 on tumor surface increased significantly by rClone30-hDR5 compared to that by rClone30, which contributed to the sensitivity to TRAIL. In conclusion, rClone30-hDR5 in combination with TRAIL has potential application value in cancer treatment.

Objective To study the methods and clinical effect of lateral less invasive stabilization system(LISS)plate combined with medial buttress plate in distal femoral fractures patients with comminuted medial cortex.Methods The clinical data of 25 distal femoral fractures patients with comminuted medial cortex from June 2014 to December 2015 were retrospectively analyzed, and all patients were treated with lateral LISS plate combined with medial buttress plate. According to the AO fracture classification,A3 type was in 10 cases,C2 type was in 9 cases,and C3 type was in 6 cases.The knee joint function at 3,6 and 12 months after operation was evaluated according to American hospital for special surgery (HSS) knee joint function score, and the clinical effect was evaluated 12 months after operation.Results The patients were followed up 18-36 months,with an average of 23.4 months.The operation time was (80 ± 22) min, intraoperative bleeding was (395 ± 56) ml, and the average hospitalization time after operation was 5.6 d.All patients got bone union,and the bone union time was 16-30 months,with an average of 18.7 months.All patients had no incision infection,no fracture and delayed union.The HSS knee joint function scores at 3,6 and 12 months were(84.3 ± 7.8),(89.3 ± 7.2) and(97.9 ± 4.3)scores.The incidence of complications was 24%(6/25),and the excellent and good rate was 84%(21/25).Patients felt satisfied.Conclusions The clinical effect of lateral LISS plate combined with medial buttress plate in distal femoral fractures patients with comminuted medial cortex is good.

In order to enhance the antitumor efficacy of recombinant Newcastle disease virus, rNDV-IL15 was rescued in this study. Recombinant plasmid prNDV-IL15 was constructed, and BHK21 cells were transfected with the recombinant plasmid. Finally, the recombinant Newcastle disease virus rNDV-IL15 was successfully rescued. The growth curves of these two recombinant viruses were determined. Murine melanoma B16F10 cells were infected with rNDV-IL15 at MOI of 0.1, and the expression level of IL15 in the supernatant was detected by ELISA. The antitumor efficacy of rNDV-IL15 and rNDV was compared in vitro and in vivo. Results showed that prNDV-IL15 was constructed and recombinant virus rNDV-IL15 was successfully rescued. The growth curve of rNDV-IL15 showed that the growth of rNDV-IL15 had not been changed after insertion of IL15 gene. Results showed that there was high level of IL15 expression in the supernatant of rNDV-IL5-infected B16F10 cells (1 044.3 +/- 27.7 ng x mL(-1)). rNDV-IL15 and rNDV significantly inhibited the growth of B16F10 cells in vitro in a time-dependent manner. However, there was no significant difference between them. In animal experiments, rNDV-IL15 efficiently suppressed tumor growth in vivo when compared with rNDV, and the difference was statistically significant. The results suggested that rNDV-IL15 is a more effective antitumor agent.

Objective To know the current situation of core competency of midwives in Guangzhou, and analyze the influencing factors.Methods A cross sectional survey was conducted among 256 midwives from 13 hospitals in Guangzhou to investigate their core competency.Results The total average score of core competency of midwives was (4.09±0.49). The scores of intrapartum care and antenatal care were higher,while the scores of progestational care and public health care were lower. Multivariate regression analysis showed that the core competency of midwives was influenced by several aspects,and they were work experience,work experience in midwifery,age,title,job and marital status (β′=0.771, 0.549,0.409,0.390,0.250,0.137;P<0.05).Conclusions The overall level of midwifery competency of midwives in Guangzhou is good,but their professional scope and role function need to be deepen and expanded,so as to satisfy the growing health care demand from pregnant women and their families. Managers should develop individualized and specific training and management strategies to improve midwives′ competency based on the condition of hospitals.