Objective In order to explore the roles of TLR2 and TLR4 in the hepatocyte dam- age caused hy hepatitis B virus infection,and to find whether LPS can affect the damage of hepato- cytes pre-and pos-HBV infection,we detected the changes of TLR2 and TLR4 expressions in hu- man hepatocyte lines HepG2 cells and 2.2.15 cells.Methods HepG2 ceils are most similar to normal human hepatocytes and 2.2.15 ceils are HepG2 cells infected with HBV.We selected these two cell lines to study the differences of TLR2 and TLR4 expression between HepG2 cells before and after HBV infection.In this research,both HepG2 and 2.2.15 cells were stimulated with 0?g/ml, 1?g/ml,10?g/ml,100?g/ml,1 mg/ml and 10 mg/ml LPS.Then the expression of protein of TLR2 and TLR4 were examined by immuno-histochemistry(IHC).The cnRNA of HepG2 and 2.2.15 ceils stimulated with 0?g/ml and 10 mg/ml LPS were examined by reversal transcription-pol- ymerase chain reaction(RT-PCR).Thereafter,the apoptosis of HepG2 and HepG2.2.15 cells were examined by flow cytometry(FC),and the expressions of HBsAg and HBeAg of HepG2.2.15 cells tested with Abbott kits.Results IHC and RT-PCR analysis revealed that TLR2 and TLR4 expres- sions could he detected in both HepG2 and 2.2.15 cells.Moreover,without immune activation, TLR2 and TLR4 expressions were higher in the presence of higher concentrations of LPS.FC analy sis revealed that no apoptosis detected in HepG2 ceils stimulated with LPS in this research,but apop- tosis could be detected in 2.2.15 cells when treated with the same factors.Furthermore,the apoptosis ratios increased with the increase of LPS concentrations.When concentrations of LPS were 1?g/ml, 10?g/ml,100?g/ml,1 mg/ml and 10 mg/ml,the apoptosis ratios were 1.94%,3.03%,3.50%, 3.72%,5.30%,respectively.Abbott analysis revealed that expressions of HBsAg and HBeAg of 2.2.15 cells stimulated with LPS were lower than those not stimulated with LPS.Conclusion HBV can affect the expressions of TLR2 and TLR4 in HepG2 cell lines.LPS can lead 2.2.15 cells to apop- tosis but not HepG2 cells.Although LPS cannot damage normal hepatocytes,it might aggravate hep- atocytes damage when their microenvironment was changed by HBV infection.

Objective To illustrate the expressional distribution of heat shock protein 70(HSP 70) in the kidney of acute renal injury rats caused by gentamicin.Methods One hundred and fifty rats were randomly divided into 3 groups: group A (normal group), group B [80 mg/(kg?d) gentamicin to be injected], group C [160 mg/(kg?d) gentamicin to be given]. The variations of renal pathology were observed at different time phasess by light scope and electromicroscope. Simultaneously, the expression of HSP 70 in kidney was evaluated by immunohistochemistry.Results HSP 70 distributed in epithelial cells of renal tubular in acute injury rats. The expression of HSP 70 increased markedly from the 6th hour after injection, peaked at the 12th hour and lasted for 48 hours. The expression of HSP 70 in group C was higher than that in group B(P

Objective To explore the efficiency and prognosis of hematological malignancies treated by haploidentical hematopoietic stem cell transplantation.Methods 70 patients who received haploidentical hematopoietic stem cell transplantation were analyzed retrospectively.According to tumor burden before transplantation,the patients were divided into three groups,the low tumor burden group,the mediate tumor burden group and the high tumor burden group.And then the effection of the tumor burden to survival was analyzed,and the engraftment,GVHD,infection,conditioning related toxicity,relapse and survival rate were also observed.Results The follow-up was terminated on January 1,2014.Follow-ups were performed for a median of 34.05 (7.4-83.6) months after transplantation.All patients achieved engraftments.The cumulative incidence of GVHD of grades 2-4 was 47.14 ％ (33/70) and that of grades 3-4 was 21.4 ％ (15/70).The chronic extensive GVHD was 20.0 ％ (14/70).The overall survival was 68.6 ％.Transplant-related mortality was 12.8 ％ and the relapse was 18.6 ％.The overall survivals in low tumor burden group,mediate tumor burden group,high tumor burden group were 91.67 ％,72.7 ％,33.3 ％ respectively.By SPSS 20.0,tumor burden was the high risk factor affecting the survival (low tumor group vs high tumor group,mediate tumor group vs high tumor group,low tumor group vs high tumor group,P =0.000,P =0.038,P =0.016).Conclusions Haploidentical hematopoietic stem cell transplantation in hematological malignancies is safe and effective.And for hematological malignancies with poor prognosis disease,it should be accepted the HSCT as soon as possible after remission in order to reduce the recurrence rate of malignancy.

Objective To investigate the efficacy and clinical effectiveness of the steroid therapy with the all body and local application on hyperthyroid Graves ophthalmopathy.Methods A total of 57 patients (106 eyes)suffering from hyperthyroid Graves ophthalmopathy who hospitalized in the Department of Ophthalmology of the Second Affiliated Hospital of Harbin Medical University from Mar 2008 to Oct 2011 were enrolled in the study.Of the 57 patients,there were 21 males and 36 females,aged 18 to 57(average 37.28 ± 11.63).The clinical course was from 1 month to 5 years(average 29.63 ± 15.36 months).Based on the patient's age,sex and condition,the 57 patients were randomly divided into steroid pulse therapy with all body group(group SPT,29 patients,54 eyes)and peribulbar injection of long-term steraid group(group PI,28 patients,52 eyes).The patients in group SPT were treated with 0.5-l.0 g methylprednisolone by intravenous infusion daily for 3 days,and then stopped for 7 days,the treatment was repeated once.But the total application dose of methylprednisolone was not overruned 5.0-20.0 g.After that,the patients were treated with oral prednisone 40 mg/d.The dose of prednisone was reduced gradually to 5 mg/d.The total course of group SPT was 3 months.The patients in the group PI were treated with triamcinolone acetonide in periorbital injection of 20 mg once every two weeks for 3 months.After one month treated,all patients were visited.The patients' body weights of two groups were measured before the treatment and after one month treated.The ocular signs of the patients including visual acuity,diplopia,intraocular pressure,exophthalmos and horizontal visual palpebral fissure width were observed before and after the treatment.Results In the group SPT,the average body weight [(62.83 ± 7.23)kg]after the treatment was higher than that before the treatment[(57.03 ±7.13)kg],and also higher than that of the group PI[(59.89 ± 10.67)kg,t =25.25,2.08,all P＜ 0.05]after the treatment.In the group SPT,the average intraocular pressures before and after the treatment were (13.46 ± 1.78)and (13.69 ± 1.41)mm Hg(1 nm Hg =0.133 kPa),respectively.In the group PI,the average intraocular pressures before and after the treatment were (13.52 ± 2.19) and (13.77 ± 2.03)mm Hg,respectively.There was no significant differences before and after the treatment in both gr0ups(P＞ 0.05).In the group SPT and PI,the average exophthalmos after the treatment[(18.63 ± 2.22) and (14.85 ± 2.61)mm]were lower than those before the treatment[(20.78 ± 2.24) and (20.60 ± 2.81)mm,t =9.97,29.68,all P＜ 0.05],respectively.After the treatment,the average exophthalmos in the group PI was significantly lower than that of the group SPT(t =7.79,P＜ 0.05).In the groups SPT and PI,the average averagepalpebral fissure width after the treatment[(15.00 ± 1.80) and (11.65 ± 2.45)mm]were lower than those before the treatment[(17.22 ± 1.89) and (17.44 ± 2.37)mm,t =22.74,91.24,all P ＜ 0.05].After treatment of the group PI,the average exophthalmos was significantly lower than that of the group SPT(t =8.01,P ＜ 0.05).Conclusion Compared with the steroid pulse therapy with all body,the peribullbar application of long-term hormone for hyperthyroid Graves ophthalmopathy is more effective,with lcss side effects and worthy of promotion.

OBJECTIVETo study the effect of ultrafiltration-membrane extracts of Radix Rehmanniae Praeparata (UMERRP) on theproliferation and genetic stability of bone marrow-derived mesenchymal stem cells (BMSCs) induced by cadmium chloride (CdCl2).

METHODSProtective effects on the proliferation, micronuclear rates, chromosome aberration rates, and apoptosis rates were observed by micronuclei test, karyotype analysis, and flow cytometry.

RESULTSCompared with the CdCl2 group, UMERRP with different molecular weights at 0. 8 g/L could obviously promote the proliferation (P <0. 05). Compared with the control group, micronuclear rates, chromosome aberration rates, and apoptosis rates were obviously enhanced in the CdCl2 group (P <0. 05). Compared with the CdCl2 group, UMERRP with different molecular weights could obviously decreased CdCl2 induced micronuclear rates, chromosome aberration rates, and apoptosis rates (P <0. 05). Of them, BMSC micronuclear rates and chromosome aberration rates decreased most obvious in UMERRP groups with molecular weight below 10 000 (P <0. 05). The apoptosis rate decreased most obviously in UMERRP groups with molecular weight ranging 100 000 and 200 000 (P <0. 05).

CONCLUSIONUMERRP could reduce CdCl2 induced micronuclear rates, chromosome aberration rates, and apoptosis rates.

Apoptosis ; Bone Marrow ; Cadmium Chloride ; toxicity ; Drugs, Chinese Herbal ; pharmacology ; Flow Cytometry ; Hematopoietic Stem Cells ; Humans ; Mesenchymal Stromal Cells ; Ultrafiltration

OBJECTIVEUsing olfactory event related potentials (OERP) and magnetic resonance to evaluate olfactory function in patients with posttraumatic anosmia.

METHODSTwenty four patients with posttraumatic anosmia were reviewed retrospectively. A thorough medical history, physical examination, nasal endoscopy, T&T olfactory testing, olfactory event-related potentials, brain computed tomography scan and magnetic resonance image of olfactory pathway were performed in all patients.

RESULTSSubjective olfactory testing indicated 20 of 24 patients were birhinal anosmia, 2 with right nostril anosmia and left impairment, 2 with left anosmia and right normal. No OERP were obtained in 24 (20 were birhinal, 4 was monorhinal), except 4 cases with single nostril. Magnetic resonance imaging revealed the injures to the olfactory bulbs (100%), rectus gyrus (91.7%), orbital gyrus (67%), olfactory tracts (8%) and temporal lobes (8%).

CONCLUSIONSOERP can objectively evaluate posttraumatic olfactory function, and magnetic resonance of olfactory pathway can precisely identify the location and extent of injures.

Adult ; Craniocerebral Trauma ; complications ; Evoked Potentials ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Olfaction Disorders ; etiology ; pathology ; physiopathology ; Retrospective Studies

OBJECTIVETo study the effects of Naoan tablets on brain hemodynamics and cerebral microcirculation of soft membrane.

METHODCerebral blood stream flux, resistance of blood vessels, blood pressure and heart rate were used as observation indexes in hemodynamics experiment. Artery caliber and the number of capillaries with recovered blood stream were used as observation indexes in microcirculation experiment.

RESULTNaoan tablets at dose of 0.5 g x kg(-1) and 1.0 g x kg(-1) could enhance cerebral blood stream flux, decrease resistance of blood vessels, and reduce blood pressure. While no effects on heart rate. Naoan tablet at dose of 0.7 g x kg(-1) and 2.1 g x kg(-1) could increase the number of capillaries with recovered blood stream and enlarge the artery caliber of soft membrane in rats.

CONCLUSIONNaoan tablets can improve the indexes of hemodynamics and cerebral microcirculation of soft membrane.

Anesthesia ; Animals ; Brain ; blood supply ; Cerebrovascular Circulation ; drug effects ; Codonopsis ; chemistry ; Dogs ; Drug Combinations ; Drugs, Chinese Herbal ; pharmacology ; Female ; Ligusticum ; chemistry ; Male ; Microcirculation ; drug effects ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Wistar ; Tablets ; Vascular Resistance ; drug effects

AIMIn order to seek the marks of the genes, the relation between the influence of endurance training on aerobic ability and ACE Gene I/D Polymorphisms were studied.

METHODS102 army recruits of Han nationality from North China for an 18 week en durance training of 5000m distance. Their VO2(max), VT and the left ventricular structure and function were measured before and after the training. We also tested their ACE Gene I/D Polymorphisms with PCR-AFLP method.

RESULTSThe compliance of VO2(max), VT and left ventricular structure and function had improved after the training; the deltaVO2(max) of ID and II type was obviously higher than that of DD type (P < 0.05); there was obviously diference of deltaVO2(VT) in different ACE genotype (P < 0.05), the deltaVO2(VT) of type II was obviously higher than that of DD type (P < 0.05).

CONCLUSIONI allele has obviously hereditary advantage on the sensitivity to aerobic training in VO2(max) and VT, and type II has relation on the sensitivity to aerobic training in VT; there is no relation between I/D polymorphism and the sensitivity to aerobic training on the structure and function of left ventricle.

Adolescent ; Asian Continental Ancestry Group ; genetics ; China ; Humans ; Male ; Peptidyl-Dipeptidase A ; genetics ; Physical Endurance ; genetics ; Polymorphism, Genetic ; genetics ; Ventricular Function, Left ; physiology ; Young Adult

OBJECTIVETo obtain highly purified tetanus toxin fragment C (TTC) with good immunogenicity.

METHODSThe gene fragment encoding TTC was amplified from Clostridium tetani plasmid DNA by PCR, inserted into the vector pET43.1a (+) and expressed in E. coli BL21(DE3)plysS. After purification using Ni2+-chelate affinity chromatography, the expressed fusion protein was digested by thrombin and the resultant TTC protein was purified with Ni2+-chelate affinity chromatography followed by identification with SDS-PAGE and Western blotting. The purifed TTC protein was then used to immunize mice to test its immunogenecity.

RESULTSThe 1373-bp gene fragment encoding TTC was obtained, and the constructed recombinant expression vector pET43.1a (+)-TTC was successfully expressed in E. coli BL21(DE3)plysS. SDS-PAGE identified a recombinant fusion protein with relative molecular mass (Mr) of 117 000, which accounted for 22% of the total bacterial protein. The TTC protein with Mr of 50 000 was obtained after purification of the thrombin digestion products of the fusion protein, with a purity reaching 95.5%. Both the fusion protein and TTC protein could be recognized by anti-tetanus toxin antibody as shown by Western blotting. The titer of the anti-serum from mice immunized with the TTC protein was 1:25 600, and the anti-serum could specifically bind to tetanus toxin.

CONCLUSIONHighly purified and immunogenetic TTC protein has been successfully obtained, which provides a good model antigen for studying antigen presentation and immune responses in vivo.

Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; Peptide Fragments ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Tetanus Toxin ; biosynthesis ; genetics ; immunology ; Tetanus Toxoid ; immunology

Two new labdane diterpenoids were isolated from 95% ethanol extract of the leaves of Callicarpa formosana Rolfe by using silica gel column, MCI column, ODS column and HPLC. Their structures were elucidated by HR-ESI-MS, NMR and ECD spectral data. All of them are new compounds, named 13E-6β-hydroxylabda-8(17),13-dien-15-oic acid (1) and 13E-7α-hydroxylabda-8(17),13-dien-15-oic acid (2). Compounds 1 and 2 were tested for antioxidant activity, and none of them had obvious activity.