Acupuncture Therapy ; Adult ; Humans ; Male ; Myelitis ; complications ; therapy

Objective To investigate the role and mechanism of B7-H4, a negative costimulatory molecule, in mediating the immunomodulatory effects of mesenchymal stem cells C3H10T1/2 (C3H10) on T cell polarization. Methods The lentiviral vectors that carried the shRNA targeting mouse B7-H4 were transfected into mouse mesenchymal stem cells (C3H10-B7-H4). The cells were co-cultured with PHA-acti-vated mice spleen lymphocytes before and after the transfection. ELISA was performed to detect the concen-trations of cytokines in supernatants of cell culture in order to elucidate the effects of B7-H4 expressed by C3H10 on T cell polarization. A mouse model of experimental allergic encephalitis (EAE) was established. Fifty C57BL/6 mice were divided into five groups including control group, EAE group, C3H10 group (injec-ting EAE mice with C3H10 cells), C3H10-NC group ( injecting EAE mice with C3H10-NC cells) and C3H10-B7-H4 group (injecting EAE mice with C3H10-B7-H4 cells). ELISA was performed to detect the soluble form of IL-2, IL-17, IFN-γ and IL-4 in plasma samples. Results Knocking down the B7-H4 gene with shRNA significantly decreased the expression of B7-H4 on C3H10 cells, which weakened the inhibitory effects of C3H10 cells on the secretion of IL-2, IL-17 and IFN-γ by spleen lymphocytes. The therapeutic effects of C3H10-B7-H4 cells on mice with EAE were weakened after silencing the B7-H4 gene expression, which was manifested as higher nerve function score and earlier onset and bring forwarded peak time of EAE than those of the C3H10 group. Treating EAE mice with C3H10-B7-H4 cells was less efficient in inhibiting the expression of IL-2, IL-17 and IFN-γin plasma. However, knocking down the B7-H4 gene had no signif-icant effect on the expression of IL-4 in terms of treating EAE with C3H10 cells. Conclusion The co-inhib-itor molecule B7-H4 expressed on C3H10 cells mediated the treatment of EAE with C3H10 cells by regula-ting Th1 and Th17 effector T cells.

AIM To study the chemical constituents from the fruits of Psidum littorale Raddi.METHODS The ethyl acetate and methanol fractions of 90％ ethanol extract from P.littorale were isolated and purified by silica,Sephadex LH-20,D-101 macroporous resin and preparative HPLC column,then the structures of obtained compounds were identified by spectral data.RESULTS Twelve compounds were isolated and identified as lupeol (1),betulin (2),nigaichigoside F1 (3),2oα-hydroxybetulinic acid (4),(-)-episyringaresinol (5),(+)-syringaresinol (6),(+)-isolariciresinol-9'-O-β-glucopyranoside (7),(7S,8R)-urolignoside (8),benzyl-β-D-glucopyranoside (9),butanedioic acid (10),β-sitosterol (11),daucosterol (12).CONCLUSION Compounds 1-9 are isolated from genus Psidium for the first time.

OBJECTIVESTo screen proteins in hepatocytes interacting with hepatitis B virus surface antigen middle protein (MHBs) with yeast-two hybrid technique for studying the biological functions of MHBs.

METHODSPolymerase chain reaction (PCR) was used to amplify the gene of MHBs from the plasmid A7 containing the whole fragment of adr subtype of HBV and the PCR product was cloned into pGEM-T vector and then evaluated by sequencing. The gene of MHBs was cut by EcoRI and BamH I from pGEM-T vector and then cloned into the yeast expression plasmid pGBKT7. MHBs bait plasmid was constructed by ligating MHBs gene with yeast expression vector pGBKT7 with yeast-two hybrid system 3 and then was transformed into yeast AH109 (a type). The transformed yeast cells were mated with yeast Y187 (alpha type) containing liver cDNA library plasmid in 2 x YPDA medium. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X- alpha -gal for selecting and screening. After extracting and sequencing the plasmid from true positive blue colonies, the results were analyzed by bioinformatics.

RESULTSA pGBKT7- MHBs yeast expressed vector was successfully constructed. Two colonies were sequenced. One colony was Homo sapiens aldolase B fructose-bisphosphate, the other was a new gene with unknown function, which was named MHBs-binding protein 1.

CONCLUSIONMHBs gene was successfully cloned. Two genes of MHBs interacting proteins in hepatocytes were obtained by yeast-two hybrid system 3. Our results brought some new clues for studying the biological functions of MHBs and the mechanisms of HBV carcinogenesis.

Genome, Viral ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B virus ; genetics ; immunology ; Hepatocytes ; Humans ; Two-Hybrid System Techniques

OBJECTIVETo explore the therapeutic effect of qi benefiting blood activating method (QB-BAM) on acute exacerbation of chronic obstructive pulmonary disease (AECOPD) patients with blood stasis syndrome (BSS) by observing its effect on plasma fibrinogen (Fg) and D-dimer (D-D) levels.

METHODSSixty AECOPD patients with BSS were randomly assigned to the treated group and the control group, 30 in each group. All patients received conventional therapy for AECOPD. Those in the treated group were additionally injected with Shengmai Injection and Tanshinone IIA Injection. Clinical efficacy and indices including levels of Fg, D-D, PaO2, and PaCO2 were measured and compared before and after treatment.

RESULTSThe effective rate was 93.3% (28/30 cases) in the treated group, higher than that of the control group [73.3% (22/30 cases) , P < 0.05]. There was no significant difference in all indices between the treated group and the control group before treatment (P >0.05). After treatment all indices were significantly improved in the two groups (P < 0.01). But in the treated group levels of Fg and D-D decreased more and levels of PaO2 increased more (P < 0.01). Plasma levels of Fg and D-D levels were negatively correlated with PaO2 (r = -0.493, r = -0.438, P < 0.01) before treatment, and also negatively correlated with PaO2 (r = -0.452, r = -0.325, P < 0.01, P < 0.05) after treatment, but they were not significantly correlated with PaCO2 before and after treatment (P >0.05).

CONCLUSIONSQBBAM could play a therapeutic role in improving prethrombotic states of AECOPD patients with BSS. Plasma levels of Fg and D-D were related to the severity of AECOPD.

Acute Disease ; Drugs, Chinese Herbal ; therapeutic use ; Fibrin Fibrinogen Degradation Products ; Fibrinogen ; Hemostatics ; Humans ; Plasma ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; Qi

OBJECTIVETo investigate the effect of hyperbaric oxygen (HBO) treatment on the expression of nitric oxide synthase (NOS) mRNA in cortex after acute traumatic cerebral injury, and to study the mechanism of HBO on brain injury.

METHODSAcute traumatic brain injury model was established with rest received free fall injury method in SD rats. 0.25 MPa HBO treatment was used 1 h or 12 h after brain injury and the cortex was isolated 6 h or 24 h after brain injury respectively. The expression of mRNA coding for nNOS, eNOS or iNOS were assayed using reverse transcription polymerase chain reaction (RT-PCR).

RESULTSThe expression of nNOS, eNOS and iNOS mRNA were significantly decreased in 0.25 MPa HBO treatment groups than those in acute cerebral injury groups (P < 0.01). The amount of nNOS, eNOS and iNOS mRNA was significantly lower in HBOT 24 h group than those in HBOT 6 h group (P < 0.05, P < 0.01). There was no significantly difference among nNOS, eNOS and iNOS mRNA in 0.25 MPa normoxic hyperbaric nitrogen groups and acute cerebral injury groups (P > 0.05).

CONCLUSIONHBO may exert significant effects on the expression of nNOS mRNA/iNOS mRNA and protect cortical neuronal from traumatic cerebral injury.

Animals ; Brain Injuries ; metabolism ; therapy ; Female ; Hyperbaric Oxygenation ; Male ; Nitric Oxide Synthase ; genetics ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley

BACKGROUNDThe experimental studies of venous thromboembolism (VTE) as an entity and the response of the pulmonary arterial endothelium after VTE are still rare. The objective of this study was to observe changes in the pulmonary arterial endothelium using a novel rat model of VTE.

METHODSRats were allocated to the VTE (n = 54) or control groups (n = 9). The left femoral vein was blocked using a microvessel clip to form deep vein thrombosis (DVT). One, four or seven-day-old thrombi were injected into the right femoral vein to induce DVT-pulmonary thromboembolism (DVT-PTE). The rats were sacrificed 1, 4 or 7 days later (D(n(1,4,7)) P(n(1,4,7)) subgroups (n = 6)), and the lungs were examined using light and electron microscopy.

RESULTSOn gross dissection, the rate of DVT formation was higher on day 1 (D(1)P(n): 100%, 18/18) than day 4 (D(4)P(n): 83%, 15/18; χ(2) = 5.900, P = 0.015) or day 7 (D(7)P(n): 44%, 8/18; χ(2) = 13.846, P = 0.000). On gross dissection, the positive emboli residue rate in the pulmonary arteries was lower in the D(1)P(n) subgroup (39%, 7/18) than the D(4)P(n) (73%, 11/15; χ(2) = 3.915, P = 0.048) and D(7)P(n) subgroups (100%, 8/8; χ(2) = 8.474, P = 0.004); however, light microscopy indicated the residual emboli rate was similar in all subgroups. Hyperplasia of the pulmonary arterial endothelium was observed 4 and 7 days after the injection of one-day-old or four-day-old thrombi. However, regions without pulmonary arterial endothelial cells and intra-elastic layers were observed one day after injection of seven-day-old thrombi.

CONCLUSIONSThis novel model closely simulates the clinical situations of thrombus formation and is ideal to study pulmonary endothelial cell activation. The outcome of emboli and pulmonary arterial endothelial alterations are related to the age and nature of the thrombi.

Animals ; Disease Models, Animal ; Endothelium, Vascular ; pathology ; Pulmonary Artery ; pathology ; Pulmonary Embolism ; pathology ; Rats ; Rats, Sprague-Dawley ; Venous Thromboembolism ; pathology

OBJECTIVETo analyze the relationship between the number of removed axillary lymph nodes and prognosis of axillary node-negative breast cancer.

METHODSThe clinicopathological data of 655 patients with breast cancer were analyzed retrospectively. The disease-free survival curves were generated according to the number of removed axillary lymph nodes using Kaplan-Meier plots. The correlation between the co-variables and rate of breast cancer-related events was analyzed using Cox model.

RESULTSThe overall five year-disease free survival rate of the 655 cases was 94.4%. The rate of patients with lymph node number ≤ 12 was 90.3%, and that of lymph node number > 12 was 96.5%, with a statistically significant difference (P = 0.009). Significantly less breast cancer-related events were observed in patients with lymph node number > 12 (15/426, 3.5%) than that in patients with lymph node number ≤ 12 (22/229, 9.6%) (P = 0.009).

CONCLUSIONSWhen axillary node dissection is indicated, dissection of lymph nodes >12 leads to much less breast cancer-related events than that in patients with dissected lymph node ≤ 12. The more lymph nodes are dissected, the more accurate prognosis can be estimated.

Adult ; Axilla ; Breast Neoplasms ; pathology ; surgery ; Carcinoma, Ductal, Breast ; pathology ; surgery ; Carcinoma, Intraductal, Noninfiltrating ; pathology ; surgery ; Disease-Free Survival ; Female ; Follow-Up Studies ; Humans ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Mastectomy ; methods ; Middle Aged ; Proportional Hazards Models ; Retrospective Studies ; Survival Rate

OBJECTIVETo examine the correlation between the mRNA and proteins expressions of gastrin(GAS), and the association of protein expression of GAS with apoptosis index(AI) and apoptosis regulation gene Fas/FasL, caspases in colorectal cancer.

METHODSThe expressions of GAS mRNA in tumor tissues of 79 cases with colorectal cancer were detected by nested RT-PCR. Cell apoptosis was detected by molecular biology in situ apoptosis detecting technic(TUNEL). Protein expressions of GAS, Fas/FasL, and caspases were detected by immunohistochemical staining (SP method).

RESULTSThe positive correlation was found between the mRNA and proteins expressions of GAS(rGAS=0.99, P<0.01). The mRNA and protein expressions of GAS in well and moderately differentiated cancers were significantly lower than those in poorly differentiated cancers (chi(2)(high vs low)=10.47, 10.23, P<0.01, chi(2)(middle vs low)=6.68, 4.95, P<0.05). The mRNA and protein expressions of GAS in papillary and tubular adenocarcinomas were significantly lower than those in mucinous adenocarcinomas, signet-ring cell carcinoma and undifferentiated carcinoma (chi(2)(papillary vs mucinous and signet-ring)=4.80, 6.22, chi(2)(papillary vs undifferentiation)=5.44, 8.43, chi(2)(tubular vs mucinous and signet-ring)=4.40, 4.38, chi(2)(tubular vs undifferentiation)=4.92, 6.43, P<0.05, respectively). The mRNA and protein expressions of GAS in Dukes' stages A, B were significantly lower than those in Dukes stages C, D (chi(2)=4.84, 4.45, P<0.01). The AI in GAS high and moderate expression groups of colorectal cancer were significantly lower than that in low expression group (q(high vs low)=6.71, q(middle vs low)=4.60, P<0.01). The positive expression rate of FasL was significantly different among GAS high, moderate and low expression groups of colorectal cancer (chi(2)=9.35, P<0.01). The positive expression rate of FasL in GAS high and moderate expression groups was higher than that in low expression group (chi(2)high vs low=6.24, chi(2)(middle vs low)=4.74, P<0.05).

CONCLUSIONSGAS plays an important role in the regulation of cell apoptosis in colorectal carcinoma, whose mechanism may be related to the aberrant expression of Fas/FasL. GAS will be one of the indicators of the biological behavior in colorectal carcinoma.

Adult ; Aged ; Caspases ; metabolism ; Colorectal Neoplasms ; metabolism ; pathology ; Fas Ligand Protein ; metabolism ; Female ; Gastrins ; metabolism ; Humans ; Male ; Middle Aged ; RNA, Messenger ; metabolism ; Young Adult ; fas Receptor ; metabolism

AIMTo study the effect of chimonin on chronic hypoxia and hypercapnic pulmonary hypertension and to explore its mechanism.

METHODSSD rats were randomly divided into normal control group (A), hypoxic hypercapnic group(B), hypoxic hypercapnia + chimonin group (C). HO-1 and HO-1 mRNA was observed in pulmonary arterioles of rats by the technique of immunohistochemistry and in situ hybridization.

RESULTS(1) mPAP was significantly higher in rats of B group than that of A and C group. Differences of mCAP were not significant in three groups. (2) Blood CO concentration was significantly higher in rats of B group than that of A group, it was significantly higher in rats of C group than that of B group. (3) Light microscopy showed that WA/TA (vessel wall area/total area), SMC (the density of medial smooth muscle cell) and PAMT (the thickness of medial smooth cell layer) were significantly higher in rats of B group than those of A and C group. (4) Electron microscopy showed proliferation of medial smooth muscle cells and collagenous fibers of pulmonary arterioles in rats of B group, and chimonin could reverse the changes mentioned above. (5) HO-1 and HO-1 mRNA in pulmonary arterioles was significantly higher in rats of B group than that of A group, they were significantly higher in rats of C group than that of B group.

CONCLUSIONChimonin can inhibit hypoxic hypercapnia pulmonary hypertension and pulmonary vessel remodeling by further increasing the expression of HO-1 mRNA.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Heme Oxygenase (Decyclizing) ; metabolism ; Hypercapnia ; metabolism ; pathology ; physiopathology ; Hypertension, Pulmonary ; metabolism ; pathology ; physiopathology ; Hypoxia ; metabolism ; pathology ; physiopathology ; Male ; Rats ; Rats, Sprague-Dawley