Objective:To express prokaryotically GII.4 human norovirus (HuNoV) VP2 protein and to prepare polyclonal antibody against VP2.Methods:Design specific primers to amplify the VP2 gene of GII.4 HuNoV, digest and connect to the prokaryotic expression vector pGEX-6P-1, transform the correctly identified recombinant plasmid into BL21 ( DE3) competent cells.Pick out and shake the monoclonal bacteria, and add IPTG to induce recombinant GST-VP2. The fusion protein was expressed, purified by GST affinity chromatography and digested to obtain GII.4 HuNoV VP2 protein. The relative molecular mass (Mr.×10 3) of the purified HuNoV VP2 protein was analyzed by SDS-PAGE. BALB/c mice were immunized with purified GII.4 HuNoV VP2 protein (0.5 mg/ml) to prepare polyclonal antibodies. Results:The VP2 protein of GII.4 HuNoV was successfully expressed and purified, with a relative molecular mass (Mr.×10 3) of about 29; the VP2 polyclonal antibody of GII.4 HuNoV was successfully prepared and its titer was as high as 1∶1 280 000. Western blot and indirect ELISA analysis showed that the polyclonal antibody could specifically bind to the VP2 antigen of GII.4 HuNoV. Conclusions:The purified GII.4 HuNoV VP2 after prokaryotic fusion expression can be used to prepare high titer polyclonal antibody.

Objective:To explore the effect of electromyography biofeedback therapy combined with oxiracetam on peripheral blood heme oxidase-1 (HO-1), soluble apoptotic molecules and cognitive function in patients with senile vascular dementia (VaD).Methods:One hundred and fourteen elderly patients with VaD from May 2018 to May 2020 in Xingtai Third Hospital were selected and divided into two groups according to the random number table method, with 57 cases in each group. Both groups were given conventional treatment. On this basis, the control group was given oxiracetam, and the observation group was given electromyography biofeedback therapy combined with oxiracetam. The treatment effects after treated for 1 month was compared between the two groups. The levels of serum HO-1, soluble apoptotic molecules sFas, sFasL before and after treatment were compared between the two groups. Cognitive function evaluated by Wechsler Memory Scale (WMS), Mini Mental State Examination (MMSE). The scores of Chinese Stroke Scale (CSS), Ability of Daily Living (ADL) before and after treatment and adverse reactions were compared between the two groups.Results:After treated for 1 month, the total effective rate in the observation group was higher than that in the control group: 93.0%(53/57) vs. 77.2%(44/57), the difference was statistically significant ( P<0.05). After treated for 1 month, the level of serum HO-1 in the two groups was higher than that before treatment, and the level of serum HO-1 in the observation group was higher than that in the control group: (30.21 ± 4.05) μg/L vs. (24.19 ± 3.47) μg/L, the difference was statistically significant ( P<0.05). The levels of serum sFas and sFasL in two groups after treatment were lower than those before treatment, and the levels of serum sFas and sFasL in the observation group were lower than those the control group after treatment: (81.57 ± 16.23) ng/L vs. (118.49 ± 25.09) ng/L, (135.47 ± 24.41) ng/L vs. (200.71 ± 30.29) ng/L, the differences were statistically significant ( P<0.05). After treated for 1 month, the CSS scores in the observation group was lower than the control group: (13.48 ± 2.15) scores vs. (17.22 ± 3.02) scores; the WMS, MMSE, and ADL scores in the observation group were higher than those in the control group: (97.75 ± 10.27) scores vs. (88.43 ± 9.16) scores, (23.82 ± 2.50) scores vs. (21.38 ± 2.19) scores, (60.16 ± 6.24) scores vs. (51.29 ± 5.52) scores, the differences were statistically significant ( P<0.05). There was no significant difference in the incidence of adverse reactions between the two groups ( P>0.05). Conclusions:Electromyography biofeedback therapy combined with oxiracetam has a significant effect in the treatment of elderly patients with VaD. It can significantly improve vascular endothelial function, regulate apoptosis factors, strengthen cognitive function, promote recovery of nerve function and daily living ability without increasing adverse reactions.

It is still at the initial stage for basic-level military doctors to participate in the standardized residency training. While, their professional backgrounds and training objectives are significantly different from those civilian residents, it is necessary to explore the standardized training methods in line with the military career demands for those basic-level military doctors. Based on the actual needs of grass-roots troops and military medicine, this study makes significant progress in the integration of standardized residency training and field surgery training methods, making the basic-level military doctors achieve the goal of field surgery training in the course of clinical practice of residents. Not only the surgical capability of basic-level military doctors, but also the medical support capability in grass-roots troops has been improved. This article summarizes the application methods and experience of field surgery teaching in the standardized residency training of military medical residents at the grass-roots level.

Objective To evaluate the safety and efficacy of microwave tissue coagulation assisted laparoscopic hepatectomy for the treatment of hepatic tumors.Methods 29 patients underwent laparoscopic hepatectomy using microwave liver tissue coagulation from December 2011 to June 2014 in the General Air Force Hospital.All patients underwent intraoperative ultrasound guided microwave coagulation assisted laparoscopic local hepatectomy.Intraoperative blood loss,operation time,postoperative hospital stay and complications were observed.Results All the operations were successfully performed without conversion to open surgery.There were 18 cases of hepatocellular carcinoma (HCC),2 cases of intrahepatic cholangiocarcinoma,2 cases of hepatic metastases from the rectal carcinoma,5 cases of hepatic hemangioma,1 case of hepatic focal nodular hyperplasia and 1 case of hepatic cystadenoma.Drainage tubes were placed in 3 patients and removed 5-7 d after the operation.The operation time was 150 (125-210) miu,the median amount of intraoperative bleeding was 120 (30-250) ml,the postoperative hospital stay was 7 (5-10) d.All patients recovered well after the operation and no complications such as intraabdominal bleeding and bile leakage were observed.The cutting edges of liver cancer of these 22 cases were negative.The postoperative follow-up time of these 22 cases were 3-36 months and no tumor recurrence and metastasis were observed.Conclusions Laparoscopic liver resection using intraoperative ultrasound guided microwave hepatic tissue coagulation can effectively control the bleeding,improve the safety of laparoscopic liver resection,which makes it a safe and effective minimally invasive technique.

OBJECTIVE: To investigate the variation on different regions of the forearm when taking the allergen skin prick tests(SPTs). METHOD: Forty-one patients with hyper-reactive nasal symptoms were included. SPTs were done on the proximal, telecentric, middle region of right forearm and telecentric region of left forearm with house dust mite and histamine. The average diameter of wheal and the reactive grade were calculated and analyzed. RESULT: The diameters of the wheal on equal position of two forearms were similar (P > 0.05). The intension of SPTs reaction decreased from proximal to telecentric region of the same forearm (P < 0.05). CONCLUSION The intension of SPTs reaction on different region of forearm are quite different, which should be noticed in clinical practice.
Adolescent ; Allergens ; immunology ; Female ; Forearm ; Humans ; Male ; Rhinitis, Allergic, Perennial ; diagnosis ; immunology ; Skin Tests ; methods ; Young Adult

Objective To investigate the significance of cortactin in primary liver cancer. Methods Fiftythree paraffin embedded primary liver cancer specimens were collected at General Hospital of Air Force of PLA from January 2002 to May 2008. The expression of cortactin was detected by immunohistochemistry, and the relationship between cortactin expression and clinicopathologic parameters was analyzed. All data were analyzed via Wilcoxon rank sum test and Spearman rank correlation analysis. Results There was a significant difference in cortactin expression among tumor capsule integrity, TNM staging, portal vein tumor thrombus and extrahepatic metastasis (u =2. 19, 3. 584, 2. 796, P ＜ 0.05 ). There was a positive correlation between tumor invasion and cortactin expression ( r = 0. 5794, P ＜ 0.05 ). Conclusions Overexpression of cortactin may be one of the factors enhancing the invasion of primary liver cancer. The level of cortactin expression can be used in evaluating the invasive potential of primary liver cancer.

BACKGROUND:The key of stem cells for treating nervous tissue injury is the transplantation of stem coils that have regeneration capacity.The structure and function of central nervous system were re-established by multiple action mechanisms.OBJECTIVE:To explore the effects and mechanisms of bone marrow mesenchymal stem cells(BMSCs)locally transplanted into rats with spinal cord injury on neurological recovery.METHODS:BMSCs were separated with density gradient centrifugation and cell attachment.10 mg/L BrdU was used for labeling before cell transplantation.Adult female Wistar rats were used to establish spinal cord injury models using an aneurysm clip,and they were then randomly divided into control group,saline group and transplantation group.In the transplantation group,BMSCs were transplanted into the damaged spinal cord by stereotaxis at day 7 following damage.In the saline group,an equal volume of saline was utilized.In the control group,the rats were left intact.Basso,Beattie and Bresnahan(BBB)locomotor rating scale was used before and at 7,14,30,60 and 90 days following damage.Rats were sacrificed at day 90.BrdU-positive cells,Brdu+neuron specific enolase,Brdu+glial fibrillary acidic protein(GFAP),Brdu+basic fibroblast growth factor(bFGF),and Brdu+brain-derived nerve growth factor(BDNF)immunohistochemistry double-staining cells and simple staining positive calls were observed.RESULTS AND CONCLUSION:The recovery of BBB function score was better in the transplantation group than in the control group(P＜0.05).The recovery speed of BBB function score was slower in the saline group than in the control group at 30 days following damage(P＜0.05).No significant difference was determined at day 90 compared with the control group(P＞0.05).BrdU-positive cells and double-staining cells of immunohistochemistry could be found at the center of damage site and 1 cm from caudal end to damaged site in rats of the transplantation group.The number of NSE,GFAP,bFGF and BDNF simple staining cells was significantly greater in the transplantation group than in the control and saline groups(P＜0.05).Results indicated that BMSC transplantation can improve the recovery of nervous function of rats with spinal cord injury.Its mechanism may be correlated with the differentiation of transplanted cells into neuron-like and glial cell-like cells,secretion or promoting secretion of neurotrophic factors in host.

Objective To investigate the expression and the early diagnostic significance of heat shock protein 70 in acute allograft rejection of liver-transplamed rats. Methods The model of rat orthotopic liver transplantation was made by using a modified "two-cuff technique". The rats were randomly divided into 3 groups. For each group, donors and receptors all included 15 rats respectively. The control group: Wistar to Wistar liver transplantation; The untreated group: SD to Wistar liver transplantation, not receiving any immunosuppressant after liver transplantation; The treatment group: SD to Wistar liver transplantation, receiving intramuscular injection of tacrolimus (FKS06, 2mg kg-1. day-1) after operation. Five rats were executed randomly in every group on the post-transplantation day 3, 5 and 7 and the graft samples were obtained for optical microscopic observation. The expression of HSPT0 in grafts was detected by using immunohistochemical method and RT-PCR. The correlation between acute rejection following liver transplantation and the expression of HSP70 in grafted liver was studied. Results There was no acute rejection examined in the control group. The untreated group showed typical allograft rejection and the rejection activity index (RIA) went up gradually after the operation (P<0.01). The treatment group showed no rejection or borderline allograft rejection. The level of HSP70 was increased transiently after operation, then reduced in the control group (P<0.05). The level of HSP70 in the untreated group was higher than in the control groupand gradually increased with the prolongation of time after transplantation (P<0.01). A significant correlation was found between HSP70 and pathological score in the untreated group (P<0.01). The treatment group showed low levels of HSP70 of all the time. Conclusions The expression of HSP70 in grafts is closely related to the occurrence and development of the acute rejection and can be useful for early diagnosis of acute allograft rejection following liver transplantation.

Objective: To explore the parameters used for the clinical evaluation of post-trau- matic hepatic function crashing down to the ground. Methods: By establishing the animal model of liver impact injury in simulation of parachuter,blood were sampled before injuries and 15 min, 30 min,45 min,1 h, 3 h, 6 h ,9 h after injuries. Cytokines including TNF?、IL-2 and liver function parameters including ALT,AST,LD-L,GGT,ALP were measured and analyzed statistically. Results: TNF? increased after 15 min- utes, and IL-2 increased after 30 minutes(P0.05). These two pa- rameters were significantly increased all the time after injuries(P

Objective To develop a rapid donor liver harvesting method in orthotopic liver transplantation in rats,and to study the stability and the success rate of the model.Methods A total of 200 healthy adult male SD rats were randomly divided into two groups: the experimental group(rapid donor liver harvesting transplant group),and the control group(conventional liver transplant group).The operation time,cold(ischemic) time and warm ischemic time of the donor liver, recipient post-transplation liver function and the success rate of the operation between the 2 grous were cpmpared.Results In the control group,the(operation) time of donor harvesting was(33.8?4.2) min,warm ischemic time and cold ischemic time of donor liver was(3.5?1.2) min and(62.0?4.2) min,with a 80% rate of success.The new method reduced the time for donor surgery to(13.1?2.2)min,and reduced the warm ischemic time and cold(ischemic) time of donor liver to 0 min and(38.0?3.1)min respectively,and with a 94% rate of success(all P