Objective To overcome the fact that SRV-NM virus can only multiple and amplify through partially pu-rified jaagsiekte retrovirus inoculated intratracheally in sheep but it cannot be augmented using in vitro cell culture, we con-structed JSRV-NM pseudovirions based on high efficiency packing system of lentivirus. Methods Lentivirus of three high efficiency packing plasmids system pMD.G, pCMV-HIV 8.2 and pHIV-eGFP was developed, and JSRV-NM-env coated plasmid pCMVJSRV-NM was used to substitute VSV-G virus coated plasmid pMD.G then co-transfected into 293T cells to replicate, package and produce restructured JSRV-NM pseudovirions. Gene expression of pseudovirion was determined through WPRE using real time PCR; Virus infectivity was detected through inoculating JSRV-NM pseudovirions into 24 pore plates. Results We construct JSRV-NM pseudovirions successfully based on the lentivirus system. JSRV-NM pseudo-virions can also be concentrated to higher titer (108 TU/mL detected by real time PCR by ultracentrifugation without signifi-cant loss of activity. JSRV-NM and VSV-G pseudovirions infected on Hela cells (both MOI= 3) respectively and no obvi-ous difference were shown on their infection efficiency detected by real time PCR. Conclusion Based on lentivirus system, JSRV-NM pseudovirions can be multipled and amplified in 293T cell culture in vitro. JSRV-NM pseudovirions is stable without loss its infection activity and the requirements of biological laboratory safety II was also met. JSRV-NM pseudoviri-ons will provide a useful tool for further study of JSRV-NM-env infection across species or its induction of lung adenocarci-noma.

OBJECTIVETo explore the correlation between Pi and Shen by observing the relationship between the metabolism of aristolochic acid (AA) and mRNA and protein expression levels of organic anion transporting polypeptide (oatp) superfamily member 2a1 and 2 b1 (oatp2al and oatp2bl) in renal, small intestinal, and large intestinal tissues of Pi deficiency syndrome (PDS) model rats.

METHODSTotally 46 Sprague-Dawley (SD) rats were randomly divided into four groups, i.e., the blank group (n = 12), the PDS group (n = 22), the AA-I group (n = 6), and the PDS AA-I group (n = 6). PDS model was established by subcutaneously injecting Reserpine at the daily dose of 5 mg/kg for 16 successive days. Carotid intubation was performed in 6 rats selected from the blank group and the PDS group. Pharmacokinetics of AA-I were detected at 5, 15, 30, 45, and 60 min after gastrogavage of AA-I. AA-I concentrations in renal, small intestinal, and large intestinal tissues of 10 rats selected from the PDS group were determined. Normal saline was administered to 6 rats selected from the PDS group and the blank group by gastrogavage. Renal, small intestinal, and large intestinal tissues were collected in the AA-I group and the PDS AA-I group at 60 min after gastrogavage of AA-I. mRNA and protein expression levels of oatp2a1 and oatp2b1 in each tissue were detected using real-time polymerase chain reaction (RT- PCR) and Western blot.

RESULTSCompared with the blank group, plasma concentrations of in vivo AA-I were obviously higher in the PDS group at 15, 30, 45, and 60 min after gastrogavage of AA-I with statistical difference (P < 0.05). Plasma concentrations of AA-I were obviously decreased at 60 min after gastrogavage of AA-I; AA-I concentrations in renal and large intestinal tissues were elevated; AA-I concentrations in small intestinal tissues were obviously reduced in the PDS group. There was no statistical difference in mRNA expression levels of oatp2a1 and oatp2b1 in the aforesaid three tissues of rats between the blank group and the PDS group. Compared with the blank group, mRNA expression levels of oatp2a1 and oatp2b1 decreased in small intestinal tissues of the AA-I group, and the mRNA expression level of oatp2a1 in large intestinal tissues significantly decreased (P < 0.05, P < 0.01). Compared with the PDS group, mRNA expression levels of oatp2a1 and oatp2b1 increased in renal tissues of the PDS AA-I group (P < 0.05); mRNA expression levels of oatp2b1 increased in large intestinal tissues of the PDS AA-I group (P < 0.05).

CONCLUSIONSThe difference in AA-I metabolism might be associated with changed expression levels of oatp2a1 and oatp2b1 in renal, small intestinal, and large intestinal tissues under Pi deficiency induced loss of transportation. Shen and Dachang played important roles in substance metabolism under Pi deficiency state, which proved Pi-Shen correlated in Chinese medical theories.

Animals ; Anions ; Aristolochic Acids ; metabolism ; Drugs, Chinese Herbal ; Kidney ; Medicine, Chinese Traditional ; Organic Cation Transport Proteins ; metabolism ; Peptides ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley

ObjectiveTo study the expression changes of peripheral blood monocyte human leukocyte antigen DR (HLA-DR) in patients with severe craniocerebral injury,and investigate the correlation between HLA-DR expression and infection and prognosis.MethodsNinety patients with craniocerebral injury were selected as experimental group and were divided according to the Glasgow coma scale (GCS) score after hospitalization into experimental group 1 (GCS score 13-15 scores ),experimental group 2 (GCS score 9-12 scores) and experimental group 3 (GCS score 3-8 scores) with 30 patients each,which were moderate,medium,severe craniocerebral injury,respectively.Thirty healthy people were chosen at the same period as control group.The HLA-DR expression of experimental group was detected after 1,3,7 and 14 d of admission by flow cytometry,and the HLA-DR expression of control group was detected on the day they got physical examination.The rates of infection,cure,disability,vegetative state and mortality were counted after 30 d of admission.ResultsThe HLA-DR expressions in experimental group 1 and experimental group 2 after 1,3,7,14 d of admission were (28.11 ± 2.37),(26.45 ± 1.63),(27.75 ± 1.83),(27.15 ± 2.17) MCF and (29.34 ±2.07),(27.55 ± 1.63),(28.42 ± 1.94),(29.46 ±2.12) MCF,which had no statistical difference compared with that in control group [(29.18 ± 1.91 ) MCF](P＞ 0.05).The HLA-DR expressions in experimental group 1 and experimental group 2 after 1,3,7 d of admission and control group had statistical differences compared with those in experimental group 3 after 1,3,7 d of admission [(18.02 ± 1.78),(16.05 ± 1.97 ),(20.76 ± 1.65) MCF ] (P ＜ 0.05).The HLA-DR expressions in experimental group 1 and experimental group 2 after 14 d of admission and control group had no statistical significance compared with that in experimental group 3 after 14 d of admission [ (26.13 ± 2.15) MCF](P＞ 0.05).The infection rates of experimental group 1,experimental group 2 and experimental group 3 were 0,3.6％(1/28),82.8％(24/29),respectively,while the cure rates were 100.0％ (30/30),100.0％ (28/28),10.3％ (3/29),the disability rates were 0,0,41.4％ (12/29),the vegetative state rates were 0,0,20.7％ (6/29),and the mortality were 0,0,27.6％ (8/29).There was no statistical significance in the rates of infection,cure,disability,vegetative state and mortality between experimental group 1 and experimental group 2 (P＞ 0.05 ).While there was statistical differences in the rates of infection,cure,disability,vegetative state and mortality among experimental group 1,experimental group 2 and experimental group 3 (P ＜ 0.05).ConclusionsThe HLA-DR expression changes of patients with moderate and medium craniocerebral injury after 1,3,7,14 d of admission are not significant.The HLA-DR expression of patients with severe craniocerebral injury begins to decline from 1 d after injury,declines obviously at 3 d,increases from 7 d,returns to normal level at 14 d.The decline of HLA-DR expression in patients with severe craniocerebral injury is correlated with the infection,and predicts poor prognosis.

Objective To introduce the mini-open anterior correction under video assisted thoracosco- py for thoracic idiopathic scoliosis and report its clinical results.Methods From July 2001 to 2006,37 ca- ses of right thoracic idiopathic seoliosis were surgically corrected with mini-open anterior instrumentation. There were 4 males and 33 females with average age of 14.1 years and average Cobb angle of 56?.Fourteen cases with Lenke I A 14 cases,IB ++-++++ 14 cases,IC 9 cases.The Risser sign was ++-++++. The operative time,blood loss,instrumented levels,correction rate and loss of correction were analyzed. Results The operative time averaged 220 miu.The intraoperative blood loss averaged 320 ml.The average number of instrumented levels was 7.8.The postoperative Cobb angle was 16.8?on average with correction rate of 70%.With a follow-up of 18-36 months,the loss of correction averaged 4.6%,but no hardware complications.Conclusion The clinical results of mini-open anterior correction under thoracoscopy for tho- racic idiopathic scoliosis were satisfactory.It may minimize the complications of classical thoracotomic anterior correcion and mav reduce the high requirement for anesthesia,long operative time and high cost of thoraco- scopic anterior instrumentation.

Objective:To investigate variety pattern of expression level of TCRV?subfamilies in mononuclear cell in spleen tissue of rats with dampness pathogenic factors and normal rats by using FQ-PCR technique. Methods:32 SD rats were divided into four groups: normal group, external dampness group, internal dampness group, external and internal dampness group. Observing period was 20 days. 3 Rats were randomly selected from each group in order to exam the TCRV? subfamilies expression level. Results:The expression of TCRV?1, TCRV?7, TCRV?9 and TCRV?13 in external dampness group were higher than those in normal group (P

Objective To observe the effect of Valpar 4 system on hand injury. Methods 40 patients with hand injury were randomly divided into control group and observation group with 20 patients in each group. Routine treatment was applied in both groups while Valpar 4 system was added in the observation group. Then total active motion (TAM) of finger joints, Disability of Arm Shoulder and Hand (DASH), and Upper Extremity Function Test (UEFT) were carried out before, 2 weeks and 4 weeks after treatment. Results The TAM improved significantly in the control group 2 weeks after treatment (P<0.001). The TAM, DASH and UEFT improved significantly in the observation group (P<0.001), and the DASH and UEFT were better in the observation group than in the control group (P<0.001). 4 weeks after treatment, the TAM and UEFT improved significantly (P<0.001) in the control group, the TAM, DASH, UEFT significantly improved in the observation group (P<0.001), and were better in the observation group than in the control group (P<0.05). Conclusion Valpar 4 system can effectively improve the TAM of fingers, function of hands and upper limb, especially in terms of activities of daily living of upper limbs.

Background Recent studies have demonstrated that epicardial flow in nonculprit arteries,which has been assumed to be normal,was slowed in the setting of ST-elevation myocardial infarction (STEMI).However,the impact of primary percutaneous coronary intervention (PCI) on blood perfusion in nonculprit arteries in patients with STEMI has not been clarified.The purpose of this study was to investigate the impact of primary PCI on blood perfusion in nonculprit arteries in patients with STEMI and correlated clinical factors.Methods A total of 117 patients with anterior wall STEMI,the culprit artery being the left anterior descending artery (LAD),undergoing primary PCI (the study group) and 100 patients with normal coronary angiography (the control group) were enrolled.To observe the differences of corrected TIMI frame count (cTFC) and myocardial blush grade (MBG) before and after primary PCI in both culprit and nonculprit arteries,the left circumflex coronary artery (LCX),cTFC and MBG in the LAD and LCX were measured in the study group and control group.The study group was divided into three groups; reflow in the culprit artery group (the R group),no reflow in culprit artery group (the NR group),and no reflow in both the culprit artery and nonculprit artery group (the NRB group) according to MBG grade.The level of serum C-reactive protein (CRP),catecholamine,and fibroblast growth factor-21 (FGF21) were assayed.The clinical and angiographic characteristics were also analyzed.Results cTFC (28.1±24.3 vs.20.3±19.3,P ＜0.05) and MBG in the LCX were different in the study group compared to the control group before primary PCI.cTFC (25.2±22.3 vs.28.1±24.3,P ＜0.05) and the MBG level in the LCX were improved after successful primary PCI,but were not recovered to the normal level.Patients with no reflow in the culprit artery had a higher incidence of no-reflow in the nonculprit artery (78％ vs.19％,P ＜0.0001),and the levels of CRP ((3.29±1.31) mg/dl vs.(2.51±1.14) mg/dl vs.(2.93±1.07) mg/dl,P ＜0.05,respectively),catecholamine ((epinephrine (693.48±89.78) pg/ml vs.(398.12±93.28) pg/ml vs.(562.54±96.22) pg/ml,P ＜0.0001,respectively),and norepinephrine ((7012.43±932.47) pg/ml vs.(4012.34±814.16) pg/ml vs.(5549.03±912.65) pg/ml,P ＜0.0001,respectively)) in the NRB group were higher than those in the R group and NR group.The level of FGF21 ((0.299±0.093) ng/ml vs.(0.612±0.071)ng/ml vs.(0.428±0.074) ng/ml,P ＜0.0001 respectively) in the NRB group was lower than that in the R group and NR group.Conclusions The blood perfusion in the nonculprit artery may be impaired in patients with STEMI.Although nonculprit artery perfusion may be improved after successful primary PCI,it is still lower than that in the control group,and may be involved in inflammation and spasms.

OBJECTIVETo investigate the effect of 6-gingerol, the main active component of ginger, on hair shaft elongation in vitro and hair growth in vivo.

METHODSFirstly, Hair follicles were co-cultured with 3 different concentration of 6-gingerol for 5 days and hair elongation in three groups was measured. Secondly, The proliferative effect of 6-gingerol on DPCs was measured using MTT assay. Thirdly, the expression of Bcl-2 and Bax in DPCs were measured using Western blotting. In vivo study, the influence of 6-gingerol on hair growth in C57BL/6 rats was measured through topical application of 6-gingerol on the dorsal skin of each animal.

RESULTSThe length of hair shaft in 20 microg/ml 6-Gingerol group (0.50 +/- 0.08 mm) is less than 0 microg/ml (0.66 +/- 0.19) mm and 10 microg/ml (0.64 +/- 0.03) mm 6-Gingerol group (P < 0.05). In cell culture, compared to 0 microg/ml and 5 microg/ml 6-Gingerol, 10 microg/ml 6-Gingerol can significantly inhibited the proliferation of DPCs (P < 0.05). Along with the growth inhibition of DPCs by 6-gingerol, the Bax/Bcl-2 ratio increased obviously. In vivo study, the hair length and density decreased a lot after using 1 mg/ml 6-gingerol.

CONCLUSIONS6-Gingerol can suppress human hair shaft elongation because it has pro-apoptotic effects on DPCs via increasing Bax/Bcl-2 ratio. It might inhibit hair growth by prolonging the telogen stage in vivo.

Animals ; Catechols ; pharmacology ; Cell Culture Techniques ; Cells, Cultured ; Fatty Alcohols ; pharmacology ; Hair ; drug effects ; growth & development ; Hair Follicle ; drug effects ; growth & development ; Humans ; Mice ; Mice, Inbred C57BL ; Plant Extracts ; pharmacology ; Rats ; bcl-2-Associated X Protein ; metabolism

OBJECTIVE The present study was aimed to investigate the role of Wnt/β-catenin sig-naling in spinal VGLUT2 regulation and neuropathic pain. METHODS To elucidate the association be-tween VGLUT2 and neuropathic pain,we determined the expression and distribution characteristics of VGLUT2 in mice subjected to spared nerve injury(SNI),and then observed the effects of two VGLUT2 targeting shRNAs on mechanical allodynia and glutamate release.The effects of Wnt/β-catenin signal-ing on VGLUT2 expression and pain behavior were investigated by using Wnt agonist,Wnt1,and Wnt/β-catenin pathway inhibitor XAV939 in SNI mice.RESULTS SNI surgery induced significant up-regula-tion of VGLUT2 on postoperative days 7,14,and 21.Double immunofluorescence labeling of VGLUT2 with NeuN,MAP2,Iba-1,or GFAP showed that VGLUT2 was mainly expressed in neurons in the dor-sal horn of the spinal cord after SNI(NeuN,MAP2).Intrathecal administration of VGLUT2 shRNAs be-fore or after SNI surgery significantly decreased mechanical allodynia and glutamate release. Mean-while,Wnt1/β-catenin signaling increased significantly after SNI surgery.Over-expression of β-catenin in PC12 cells increased VGLUT2 protein level,intrathecal administration of Wnt agonist or Wnt1 signifi-cantly increased VGLUT2 protein expression in spinal cord, while Wnt/β-catenin pathway inhibitor XAV939 decreased VGLUT2 expression in PC12 cells and spinal cord.Additionally,intrathecal admin-istration of XAV939 7 days after SNI significantly attenuated mechanical allodynia in mice, which was in accordance with down-regulation of VGLUT2 protein levels.VGLUT2 shRNAs significantly attenuat-ed Wnt agonist or Wnt1 induced mechanical allodynia. CONCLUSION Wnt1/β-catenin signaling path-way up-regu-lates the spinal VGLUT2 expression,and this regulation is involved in neuropathic pain behavior.