Objective To construct a baculovirus expression vector for expression of human Ro60 cDNA. Methods Ro60 cDNA was cloned from Hela cell RNA with RT-PCR and was inserted into plasmid pFastBac HTc. Plasmid pFastBac HTc-Ro60 was then transformed into E. coli DH10Bac competent cells for transposition. The constructed recombinant was determined by both blue-white colony screening and PCR analysis. Results A 1.56kb Ro60 cDNA was obtained from Hela cell RNA and was successfully inserted to plasmid pFastBac HTc. DNA sequencing and restriction enzyme analysis revealed that both open reading frame and sequence of the cDNA were identical to Ro60 sequence previously published. Specific transposition and virus recombination were obtained in the vector Bacmid-Ro60. Conclusion Ro60 cDNA has been successfully cloned and constructed to baculovirus expression vector, which is useful for protein expression and further study of this autoantigen in the pathogenesis of lupus erythematosus.

We only prolongated the course of shock from 2h to 5h, without predi- sposing the dogs. At 5h after hemorrhagic shock, the experimental results were asfollows: The ultrastructural damages were not only detected in the lung but also in theheart, liver, intestine and kidney etc. Major pathological lesions were: swelling of paren-chymal cells in various organs, vacular degeneration, and de-connection of cells. The laterwas shown as disintegration of hepatocyte-conjunction and disappearance of microvilli. Theorganelles of various cells ruptured to different degrees, for example, mitochondria swelledand its crista dissolved, endoplasmic reticulum dilated and de-granulated. In addition, thefibril of cardiac muscle thinned and dissolved, the capillary endothelial tight-conjunctionsof lung and intestine ruptured etc. The quantity of entero-endotoxin in plasma increasedin accordance with prolongation of shock, and its quantity was most abundant in portalvein, the next was artery and the last was hepatic vein. All experimental results showedthat the entero-endotoxin was related to the changes of ultrastructure of various organs.

Objective To investigate the prognosis of two subtypes of subacute cutaneous lupus erythematosus (SCLE). Methods Forty patients with subacute cutaneous lupus erythematosus were followed-up for 3 to 14 years. The clinical and laboratory findings, the response to treatment and the prognosis in these cases were comparatively analyzed. Results During the follow-up period, the patients with annular erythema subtype of SCLE showed recurrent annular skin lesions and mild systemic manifestations. Some patients gradually developed sicca symptoms. On the other hand, most patients with papulosquamous subtype of SCLE developed systemic LE manifestions. 21.7％ of the patients in the annular erythema group had mild nephritis which could be controlled by lower dose of corticosteroids and no patients died. However, 64.7％ of the patients in the papulosquamous group had moderate to severe lupus nephritis, which required higher dose of corticosteroids or immunosuppressive agents to control. Among them, One patient died and 3 patients developed uremia. Conclusion Annular erythema subtype of SCLE is milder and has a better prognosis than that of papulosquamous subtype.

Objective To study the effect of fluoride varnish on microconsistency of enamel after under irradiation in vitro. Methods Normal enamel samples were randomly divided into four groups: control group,irradiation group,fluoride varnish group and fluoride varnish-irradiation group.The microconsistency of enamel surface before and after treatment,after anti-caries test was measured by type MH-5 durometer. Results Fluoride varnish had an inhibitory effect on the decrease of enamel microconsistency etched with lactate buffer.The microconsistency in irradiation group was obviously lower than that in control group(P

[Objective]To measure adult normal femur specimen in order to obtain accurate femoral resection and match the distal femoral in total knee arthroplasty.[Method](1)Forty-seven adult normal femur specimens of central Chinese were used for taking digital pictures in both anteroposterior and medialateral position.Five angles were measured through drawing lines of fixed point.(2)The exact position of the entry point was defined by the two wires on the specimens.[Result](1)The five angles reflect distal femoral anatomy features on the frontal plane and sagittal plane.(2)The mean value of distance from the entry point to the center of the intercondylar notch was 6.21 mm and the distance from the entry point to the anterior margin of the femoral posterior cruciate ligament(PCL) insertion is found to be 6.7 mm.[Conclusion](1)The distal one-third of femora was found to be outward bowing on frontal plane and forward bowing on Sagittal plane.Especially,the forward bowing is more remarkable than Outward bowing.So,when the position of intramedullary guide stick is decided by distal femoral anatomy,the axis alignments on frontal and Sagittal planes are both important.(2)During total knee afthroplasty the exact position of entry point may be defined by the Center of the intefcondylar notch and femoral PCL insertion.

Objective To detect the expression of the tryptase in the plasma,and study the meaning in brain traumatic patients.Methods There were two groups:the brain traumatic group(40 patients)and the control group (20 health people).The content of plasma tryptase was determined by fluorescence enzyme immunoassay..Results The level of plasma tryptase had no statistical significance in control group(2.97 ±1.05)μg/L compared with the brain traumatic group(3.03 ±1.39)μg/L,however there had statistical significance comparing with sever brain traumatic patients(3.84 ±0.52μg/L)(t =3.32,P <0.05).4 cases of death in patients with severe head injury group content of tryptase (5.85 ±1.05)μg/L,which was significantly higher than the group of 16 cases of injury in severe head injury after 2 months still alive with the content of serum tryptase (2.49 ±0.52)μg/L,the difference was statistically significant (t =8.13,P <0.01).Conclusion The plasma tryptase level in sever brain traumatic patients increased significantly,and might be of importance for treatment strategies and prognosis.

Objective To observed the effect of high-intensity focused ultrasound (HIFU) in combination of submicrobubble on ablation of breast tumor VX2 in rabbits.Methods Submicrobubble was prepared,and the appearance,distribution,diameter,and zeta potential were measured.The rabbits were injected with the tumor tissue suspension in bilateral breast tissues.HIFU(150 W,120 W,5 s,3 s) combined with submicrobubble to damage rabbit breast VX2 tumors.The coagulation necrosis was assessed by HE staining.The grayscale changes of the target area before and after HIFU irradiation were recorded automatically by computer.Results The diameter of the submicrobubble was 498.9 nm on average.HE staining showed the area of coagulation necrosis was significantly larger in the HIFU + submicrobubble group than the HIFU + PBS group.The injury range of 150 W irradiation was larger than that of 120 W irradiation,given the same irradiation time.The injury range of 5 s irradiation was larger than that of 3 s irradiation,given the same irradiation strength.The gray scale change in the target area was significantly higher in the HIFU + submicrobubble group than the HIFU + PBS group.Conclusions The submicrobubble exhibited sound imaging performance,and it can be use in the ultrasound therapeutics in the future.

Objective To explore the effects of H.pylori and crude extracted proteins secreted by H.pylori(broth culture filtrate protein,BCF-P)on acid secretion from isolated rabbit parietal cells.Methods Parietal cells from rabbit gastric mucosa were isolated and enriched with digestion and elutriation.H.pylori(NCTC 11637,CagA+ VacA+)were grown in liquid broth culture and BCF-P was precipitated with ammonium sulfate.The vacuolation activity of BCF-P was evaluated with neutral red dye uptake test in HeLa cell.Isolated parietal cells were incubated with H.pylori(bacteria/cell=100∶1)for 2 h and 16 h,or BCF-P(100μg/ml)for 1 h and 12 h.Acid secretion from parietal cells was studied using 14C-aminopyrine(14C-AP)accumulation indirectly and H+-K+ ATPase α subunit mRNA expression was assessed using RT-PCR.Results (1)BCF-P containing vacuolating cytotoxin(VacA)with vacuolation activity on HeLa cells had positive result on neutral red uptake test.(2)The basal expression of H+-K+ ATPase α subunit mRNA could be detected in isolated parietal cells and 14C-AP accumulation was significantly increased in response to the stimulation of histamine with different concentrations for 30 min(P＜0.05).These results indicated that the isolated parietal cells retain relative intact acid secretion function.(3)The histamine(1.0×104 mol/L)stimulated acid secretion was inhibited sustainedly in response to H.pylori by 81% at 2 h and by 94% at 16 h(P＜0.05).However,H+-K+ ATPase α subunit mRNA expression was up-regulated in tlle acute period(2 h)and was down-regulated in the chronic period (16 h)by H.pylori(P＜0.05).(4)BCF-P significantly inhibited the histamine-stimulated acid secretion by 24% at 1 h and by 58% at 12 h(P＜0.05),and this inhibition was accompanied by the down-regulated expression of H+-K+ATPase α subunit mRNA.Conclusions Intact H.pylori and VacA secreted by H.pylori could directly inhibit histamine-stimulated acid secretion from parietal cells and this inhibition may be mediated by the down-regulated H+-K+ ATPase expression.

BACKGROUND:The improvement of athletic achievements needs a super-limited exercise to gain,how to protect athletics health under the prerequisite of gaining excellent achievements?OBJECTIVE:To reduce the injury of athletic renal function following maximum exercise by using traditional Chinese medicinal methodDESIGN:A randomized and controlled trial,taking students majoring in sports education specialty as subjects.SETTING:Internal Medicinal Department of Traditional Chinese Medicine (TCM) Hospital of Qufu Hospital of Traditional Chinese Medicine and Sports Education College of Qufu Normal University.PARTICIPANTS :Two small natural classes were randomly drown up from 16 small natural classes of 2000 grade in Sports Education College of Qufu Normal University, with 20 students in each class, on April 20, 2004.METHODS:All 40 students were according to natural class divided as experimental class and control class.The urine specimens for test were collected 15-20 minutes after one time maximum loading training on April24,2004 and immediately after morning getting-up. The procedure was performed once more on May 24. During the process the students in experimental group had taken the capsule of kidney-and-spleen-strengthening Chinese drug (KSSD, containing mainly Rhizoma Dioscoreae, Radix Polygoni Multiflori, Herba Epimedii, Fructus Lycii, Gynostamma Pentaphyllum,Radix Acanthopanacis Senticosi,Radix Salviae Miltiorrhizae,Radix Bupleuri, etc., 0.3 g/pellet), 6 pellets, 3 times a day. Urinary glucoprotein,β2-microglobulin, albumin and globulin were detectedby radio-immunoassay.MAIN OUTCOME MEASURES:The changes of urinary proteins of the students before and after administration following maximum exercise and getting-up in next morning.RESULTS: All students involved were good for the trial, without any loss.① The contents of various urinary proteins for the subjects after the maximum exercise or before administration were all obviously increased,and the conditions of natural recovery of proteinuria on getting-up next morning were quiet different: the recovery of urinary albumin was the fattest, basically to the normal;recoveries of β2 micro-globulin and immune-globulin were obvious, but the recovery of glucoprotein was inconsiderable.② After the second trial,in comparing the experimental group with the control group it was found that KSSD can obviously influence the recovery of proteinuria following maximum exercise:The effect in 12 hours did not be shown because of too fast natural recovery of albuminuria. It could promote the recoveries of β2 micro-globulin and immune-globulin. The effect on recovery of urinary of glucoprotein was obvious,basically to the normal for experimental group and basically no change for control group,there was a significant differentiation between the two groups.CONCLUSION:The contents of urinary proteins were obviously increased following maximum loading training, and natural recoveries of the four kinds of proteins presented regulatory changes.KSSD can promote the recovery of proteinuria following maximum exercise to normal volume,suggesting that the medicine has a protecting role to renal function.

Objective To investigate the potential mechanism of interleukin-1β(IL-1β)in H.pylori-related gastric carcinogenesis.Methods Human gastric cancer cell line(AGS),human gastric epithelial cell line(GES-1)and isolated parietal cells were treated with exogenous IL-1β(10 ng/ml)in the presence or absence of H.pylori(NCTC 11637).Cell proliferation and apoptosis were determined by MTT assay and flow cytometry,respectively. Expressions of cyclooxygenase-2(COX-2)mRNA and protein were examined using RT-PCR and flow cytometry,respectively.Acid secretion by parietal cells was tested using 14C-aminopyrine(14 C-AP) accumulation indirectly.H+/K+ ATPase α subunit mRNA expression was assessed using RT-PCR.Results The cell proliferation and H.pylori-related apoptosis in both GES-1 and AGS celL lines were stimulated and inhibited by IL-1β.IL-1β induced expression and upregulation of COX-2 mRNA in GES-1 and AGS cell lines,respectively.In addition,IL-1β continuously inhibited the ability of histamine-stimulated 14C-AP accumulation of isolated parietal cells accompanied by down-regulation of H+/K+ ATPase mRNA expression.Conclusions It suggests that IL-1β play an important role in H.pylori-related gastric carcinogenesis through two pathways:①to interfere gastric epithelial cell growth by up-regulating COX-2 expression,②to inhibit acid secretion from parietal cell by down-regulating H+/K+ ATPase expression.