1.Study on the expression of a substrate-based polypeptide and its inhibition against HCV protease in vitro
Wen LIU ; Wei HU ; Shigan LING
Chinese Journal of Microbiology and Immunology 2008;28(2):119-122
Objective To synthesize an inhibitory peptide against serine protease of hepatitis C virus(HCV)and explore its inhibition ability in vitro. Methods Based on the sequence characteristics of four natural substrates of protease in HCV,the gene sequence of an inhibitory peptide against HCV serine protease was designed and directly synthesized by PCR. The segment was subcloned into prokaryotic expression vector pBVIL1,resulting in the construction of the recombinant plasmid pBVIL1/IP,which was then transformed into E. coli HB101 strain. Purified by ion exchange chromatography,the expressed protein was added into an in vitro system,which was comprised of the inhibitory peptide(expressed protein),protease and the substrate,i.e. a NS5A-B fragment,in phosphate buffer. Then SDS-PAGE was performed to test the inhibition effect of the polypeptide. Results The recombinant expression vector pBVIL1/IP containing target gene was successfully constructed. The peptide expressed as inclusion boay was identified by SDS-PAGE. The degradation of protease substrate NS5A-B fragment is inhibited proportionally with the increasing concentrations of the peptide. Conclusion The recombinant peptide shows inhibitory effect on HCV protease in vitro.
2.Role of β-endorphin in conA-induced spleen cell proliferation in rats with traumatic hemorrhagic shock
Aiqing WEN ; Liangming LIU ; Deyao HU
Journal of Third Military Medical University 2001;23(4):404-406
Objective To investigate the role of β-endorphin (β-EP) in conA-induced spleen cell proliferation after traumatic hemorrhagic shock. Methods ①Wistar rats with traumatic hemorrhagic shock were used and killed 0, 1, 3, 6,12 and 24 h after traumatic hemorrhagic shock. Plasma specimens were collected and β-EP levels in plasma were detected. Rats with sham-operation served as the control. ②Spleen cells isolated from normal rats were cultured in shock plasma (group Ⅰ), inactivated shock plasma (group Ⅱ) and shock plasma+β-EP antiserum (group Ⅲ) respectively. Con A-induced spleen cell proliferation was observed. Results ①The plasma β-EP level was elevated significantly immediately after shock, and reached the peak 1 h later, then showed a deceasing tendency and restored to the level as before shock at 24 h. ②Shock plasma remarkedly suppressed spleen cell response to the mitogen conA (P<0.01) compared with control; ConA-induced spleen cell proliferative function in group Ⅱ was significantly increased than that in group Ⅰ (P<0.01), so did in group Ⅲ, which still lower than in control. Conclusion The significantly elevated β-EP in the plasma after hemorrhagic shock might play an important role in inhibiting the proliferation of spleen cells.
3.CT features of mesenteric lymphadenopathy in active Crohn’s disease
Wen LIU ; Wei WANG ; Longlong XIE ; Yuequn HU ; Sheng LIU
Journal of Practical Radiology 2016;32(4):610-613
Objective To investigate the CT features of mesenteric lymph nodes in patients with active Crohn’s disease.Methods The CT findings in 54 patients with active Crohn's disease proved by histology were analyzed,and the anatomic distribution,size, number,shape and enhancement ratio (ER)of the mesenteric lymph node were assessed.Results Mesenteric lymphadenopathy in 38 patients (70.4%)was found with a total number of 242,83.5%(202/242)of whom were located at the mesenteric root and 16.5%(40/242) at mesenteric edge.The size of the lymph nodes at the mesenteric root was larger than that at the mesenteric edge (8.57 mm±2.26 mm versus 5.38 mm±0.1 9 mm,the mean maximum short diameter),and 73.6% (1 78/242 )of the lymph nodes were oval in shape.The lymph nodes showed significant enhancement after contrast injection with an ER of 0.53 ±0.09.Conclusion Active CD often leads to mesenteric lymphadenopathy,which is more obvious at the mesenteric root.
6.Role of cemokine ligand 21 in spinal cord in tibia bone cancer pain in rats
Jiao LIU ; Wen SHEN ; Dongmei YUE ; Liping CHEN ; Xueming HU
Chinese Journal of Anesthesiology 2012;32(4):437-439
Objective To investigate the role of chemokine ligand 21 (CCL21) in the spinal cord in tibia bone cancer pain (BCP) in rats.Methods Forty adult female SD rats weighing 160-180 g were randomly divided into 5 groups (n=8 each):sham operation group (group Ⅰ ); sham operation + CCL21 neutralizing antibody group (groupⅡ); BCP group (group [); BCP + PBS group (group Ⅳ); BCP + control IgG group (groupⅤ)and BCP + CCL21 neutralizing antibody group (group Ⅵ).BCP was induced by inoculating Walker-256 mammary gland carcinoma cells into the rat tibia medullary cavity in groups Ⅲ-Ⅵ.PBS 15 μl,IgG 10 μg and CCL21 neutralizing antibody 10 μg were injected intrathecally (IT) at 14 days after intra-tibial injection of Walker-256 mammary gland cancer cells in groups Ⅳ- Ⅵ respectively.Mechanical withdrawal threshold to yon Frey filament stimulation (MWT) was measured at 1 day before (To,baseline) ; 7 and 14 d after Walker-256 cell injection (T1,T2)and at 0.5,1,2,4,8,12,24 and 48 h after intrathecal injection (T3-10 ).Results Intra-tibial injection of Walker-256 mammary gland cancer cells significantly decreased MWT as compared with the baseline values in administration of CCL21 neutralizing antibody at T5-8 as compared with MWT before intrathecal administration at T2 in group Ⅵ.MWT was significantly lower in groups Ⅲ- Ⅳ than in groups Ⅰ and Ⅱ.MWT was significantly higher at T5-8 in group Ⅵ than in groups Ⅲ - Ⅴ.Conclu]sion CCL21 in the spinal cord is involved in the maintenance of tibia BCP in rats.
7.Non-extraction treatment of dental crowding and protrusion in prepuberty:A case report
Xiaoying HU ; Chunyan LIU ; Wen WANG ; Yinghui LI
Journal of Practical Stomatology 2014;(4):580-582
Non-extraction treatment for a case with dental crowding and protrusion in prepuberty is reported.After the slow palatal expan-sion for the treatment of dental crowding of maxilla,the patient underwent orthodontic treatment.The mandibular growth led to the correction of dental crowding and the improvement of jaw profile.
8.The effect and their mechanism of acid pocket in reflux esophagitis
Yedong HU ; Lu BAI ; Fei LIU ; Wen XU ; Wei LI
Chinese Journal of Digestion 2010;30(12):881-884
Objective To investigate the function of acid pocket in reflux esophagitis. Methods The 15 healthy controls and 24 reflux esophagitis patients were identified by reflux disease questionnaire (RDQ) and gastric endoscopy. The location of subjects' lower esophageal sphincter (LES) was determined by 4 channel esophageal manometry system. Then a single-channel pH electrode was positioned 1 cm below the distal border of the LES to monitor fasting pH for half an hour. After a standard meal, the pH was continuously measured for two hours. Then the electrode was moved to 5 cm above the proximal border of the LES to monitor the dynamic pH for 24 h.Results Acid pocket was found in 16 cases of reflux esophagitis patients(66.67%) and 10 cases of healthy individuals (10/15). Acid pocket occurred earlier in reflux esophagitis group than healthy controls [11.00(4.25-17.00) min vs 30.00(15.50-54.25) min, P<0.05], and the average pH value was lower [1.84(1.59-2. 19) vs 2.32 (1.96-2.71), P<0.05]. There was no statistic difference in mean pH value of gastroesophageal junction and the duration of acid pocket before the meal.Conclusion There is abnormal acid reflux in reflux esophagitis patients, and acid pocket with earlier occurrence and lower pH value may relevant to esophageal mucosal impairment.
9.Analysis of the Distribution and Drug Resistance of Pathogens in Our Hospital in 2015
Zhenyu LIU ; Chunlin CHEN ; Rong XU ; Wen HU
China Pharmacy 2017;28(11):1475-1478
OBJECTIVE:To provide reference for rational use of antibiotics in the clinic. METHODS:In retrospective study, the distribution,composition and drug resistance of pathogens in our hospital during Jan.-Dec. in 2015 were analyzed statistically. RESULTS:In 2015,a total of 9401 specimens were collected in microbiology laboratory of our hospital;pathogens were detect-ed in 1743 specimens with positive rate of 18.54%;1591 strains of pathogens were isolated,mainly from sputum (59.77%), urine (14.775) and blood sample (8.93%). Totally 347 strains of Gram-positive bacteria(21.81%),991 strains of Gram-negative bacteria (62.29%),253 strains of fungus (15.90%)were detected. Top 4 pathogens in the list of amount were Escherichia coli, Klebsiella pneumoniae,Pseudomonas aeruginosa and Staphylococcus aureus. S. aureus,Streptococcus pneumoniae and Staphylo-coccus haemolyticus were sensitive to vancomycin,linezolid and sodium fusidate,with resistance rate of 0;resistance rates of them to erythromycin were more than 65%. The resistance rate of S. aureus to penicillin was more than 95%,and S. haemolyticus showed high resistance rate to common antibiotics. Drug resistances of E. coli,K. pneumoniae and P. aeruginosa to carbapenems were all lower than 15%,and they were sensitive to polymyxin with resistance rate of 0. Resistance rate of E. coli to quinolones was more than 50%. CONCLUSIONS:The main pathogens are Gram-negative bacteria in our hospital,drug resistance of them are not satisfactory. Drugs most sensitive to main pathogens include vancomycin,linezolid,carbapenems,etc. The inspection of patho-gen drug resistance characteristics should be strengthened,and antibiotics should be selected rationally and normatively according to the results of drug sensitivity test.
10.Role of ?-endorphin in the suppression of cellular immunity of rats following trauma-hemorrhagic shock
Aiqing WEN ; Jun WANG ; Deyao HU ; Liangmin LIU
Chinese Journal of Pathophysiology 1986;0(01):-
AIM: To explore the role of plasma ?-endorphin(?-EP) in the suppression of cellular immunity following trauma-hemorrhagic shock(T-HS). METHODS: ① Wistar rats with T-HS were sacrificed at 0 h, 1 h, 3 h, 6 h, 12 h, 24 h after T-HS. Plasma sample was collected and ?-EP levels in plasma was measured. Rats with sham-operated were served as the controls. ②In in vitro experiment, splenic cells were isolated and mixed from four normal rats and cocultured with shock plasma (SP) or SP +?-EP antiserum. ConA-induced splenic cell proliferation, IL-2 production, IL-2R expression were examined. RESULTS: ① Levels of plasma ?-EP elevated remarkably after T-HS immediately, peaked at 1h , showed decreasing tendency and restored normal 24 h after T-HS. ② Shock plasma significantly suppressed ConA-induced splenic cell function. Levels of plasma ?-EP were negatively correlated with spenic cell proliferation and IL-2 production and IL-2R expression. Compared with SP group, splenic cell function elevated markedly in SP + ?-EP antiserum group, but still lower than that in controls. CONCLUSION: The elevated plasma ?-EP following T-HS was involved in the suppression of cellular immunity.