Objective patients with pSS and 30 cases of healthv contrels.The streptavidin immunohistochemical staining was performed to detect the expression and distribution of MMP-9 and CD147 in labial salivary glands.Quantitative analysis was performed by image analysis software-image plus 5.0 at the site of positive expression of MMP-9 and CD147.The correlation between their expression and the infiltrating lymphocyte foci per 4 mm2 of labial gland was analyzed by SPSS software as well as the correlation between the expression of MMP-9 and CD 147 in the salivary glands of patients with pSS.Results MMP-9 was hiKhly expressed in labial salivary glands from 52 patients with pSS and 30 healthy controls,but the expression of MMP-9 in pSS was stronger compared with that of healthy controls(P<0.01).There was a significant positive correlation between lymphocyte foci score and up-regulated expression of MMP-9 in labial salivary glands from 52 patients with pSS(P<0.01).CD147 was highly expressed in labial salivary glands from 52 patients with pSS and 23 healthy controls，but over-expression of CD147 in PSS was more Drominent compared with that of controls(P<0.01).There was a significant positive correlation between lymphocyte foci score and up-regulated expression of CD147 in labial salivary glands from 52 patients with pSS(P<0.01).The expression of MMP-9 and CDl47 was detected in ductal and acinar epithelial cells,lymphocyte foci in pSS.There was linear correlation between the expression of MMP-9 and CDl47 in the salivary glands of patients with pSS(P<0.01).Conclusion The results of this study suggest that the abnormal expression of MMP-9 and CD 147 is involved in the pathogenesis of pSS and play a crucial role.The interaction of MMP-9 and CD147 may be one of theimportant mechanisms leading to labial salivary glands destruction found in pSS.

OBJECTIVETo investigate the anti-tumor activity of CCL21-exCD40L eukaryotic expression vector.

METHODSCCL21-exCD40L fusion gene were constructed by overlap PCR connecting CCL21 and exCD40L through a flexible linker (Gly3Ser)4, and then was cloned into expression vector pcDNA3.1(+). pcDNA3.1(+)/CCL21 and pcDNA3.1(+)/exCD were constructed as negative control. Wsestern blot was used to identify the fusion protein. CHO cells was transfected with pcDNA3.1(+)/CCL21-exCD, pcDNA3.1(+)/CCL21 and pcDNA3.1(+), respectively. The chemotatic function of the expressed product was detected by Transwell method and its anti-tumor activity was tested with vivo transfection.

RESULTSGene sequencing and restrictive digestion proved the successful construction of pcDNA3.1(+)/CCL21-exCD40L,and its expression was conformed by western blot. The transfectant supernantes of pcDNA3.1(+)/CCL21-exCD40 group had a significant chmotactic function to DCs, of which the cell numbers passing through the film was 14.95 times of blank control every high power microscope visual field. After tumor orthotoic injection of plasmid carrying fusion gene in Balb/c mouse, the tumor mass reduced remarkablely, and all the mouse in fusion gene group survived after 4 weeks.

CONCLUSIONCCL21-exCD40L fusion protein had a remarkable function to DCs and it can inhibit tumor growth and prolong the mouse survival time, which is more effective than all control group.

Animals ; CD40 Ligand ; genetics ; pharmacology ; CHO Cells ; Cell Line, Tumor ; Chemokine CCL21 ; genetics ; pharmacology ; Colonic Neoplasms ; therapy ; Cricetulus ; Dendritic Cells ; drug effects ; physiology ; Genetic Therapy ; Mice ; Mice, Inbred BALB C ; Recombinant Fusion Proteins ; pharmacology

OBJECTIVETo construct the murine CCL21 eukaryotic expression plasmid, and to investigate the chemotactic function of its products.

METHODSMurine CCL21 cDNA was amplified by RT-PCR from murine total RNA, and was inserted into eukaryotic expression plasmid pcDNA3.1 after confirmation of sequencing. The recombinant CCL21 plasmid was transferred into mouse forestomach carcinoma (MFC) cells and the chemotactic function of expressed products was detected by chemotaxis assay.

RESULTGene sequencing, gel electrophoresis of PCR products and restrictive digestion proved the successful construction of CCL21, and its expression was confirmed by Western Blot. The transfected tumor cells had a significant chemotactic function to DC.

CONCLUSIONThe recombinant murine CCL21 eukaryotic expression plasmid has been successfully constructed, and its expression products in tumor cells have a marked chemotactic function to DC.

Animals ; Base Sequence ; Chemokine CCL21 ; biosynthesis ; genetics ; Chemotaxis, Leukocyte ; Cloning, Molecular ; DNA, Complementary ; genetics ; Dendritic Cells ; drug effects ; immunology ; Genetic Vectors ; genetics ; Mice ; Mice, Inbred C57BL ; Molecular Sequence Data ; Plasmids ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Stomach Neoplasms ; metabolism ; pathology ; Transfection ; Tumor Cells, Cultured

In this study, an approach based on triple-color fluorescence probes was developed for screening potential nephro-protective bioactive substances. Three fluorescent probes (i. e. FDA, MTR and Hoechst 33342) were used to label HK-2 cells injured by doxorubicin hydrochloride, and cellular fluorescence images were subsequently acquired and analyzed by a cellular-fluorescence image microscopy platform. The established method was applied to screening 53 components of Carthami Flos, and three components C17, C18 and C19 were found to exhibit nephroprotective effects against doxorubicin hydrochloride induced injury on HK-2 cells. Eight compounds (i. e. hydroxysafflor yellow A, 6-hydroxykaempferol-3-O-rutinoside-6-O-glucoside, 6-hydroxykaempferol-3,6-di-O-gluco-side or 6-hydroxykaempferol-6, 7-di-O-glucoside, 6-hydroxykaempferol-3-O-rutinoside, 6-hydroxykaempferol-3-O-glucoside or 6-hydroxykaempferol-7-O-glucoside, rutin, isoquercetin, and kaempferol-3-O-rutinoside) in components C17, C18 and C19 were preliminarily identified by liquid chromatography-mass spectrometry (LC-MS). Isoquercetin, rutin, kaempferol-3-O-rutinoside, and hydroxysafflor yellow A were confirmed by comparing with reference substances, Further study indicated that these four compounds had moderate nephroprotective effects, while isoquercetin showed a significant nephroprotective effect in a dose-dependent manner. These results suggest that isoquercetin, rutin, kaempferol-3-O-rutinoside and hydroxysafflor yellow A might be the nephroprotective bioactive substances in Carthami Flos.
Carthamus ; chemistry ; Cell Line ; Chromatography, High Pressure Liquid ; Dose-Response Relationship, Drug ; Drug Evaluation, Preclinical ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Flowers ; chemistry ; Fluorescent Dyes ; chemistry ; Humans ; Kidney ; chemistry ; cytology ; drug effects ; Protective Agents ; chemistry ; pharmacology

Objective To study the expression of E-cadherin( E-cad), p-catenin(β-cat) in labial salivary glands of patients with primary Sj(o)gren's syndrome (pSS) in order to explore their role in pathogenesis. Methods Biopsies of labial salivary glands were obtained from 52 patients with pSS and 30 healthy controls. The immunohistochemical staining was used to detect the expression of E-cad and β-cat. Anti-SSA and anti-SSB antibodies were measured. Semi-quantitative analysis was performed by image analysis software. Ultra-structural changes was used by electron-microscopic techniques. Results ① The area of expression, optical intensity and the accumulated optical intensity of the E-cad group [(2513±1086) μm2, 0.212±0.041, 566 ±297 ] were lower than normal controls. The expression level was reduced as the increase of lymphocyte infiltration focus. ② The area of expression, the optical density and the accumulated optical density of the β- cat group [(12 324±7883) μm2, 0.113±0.031, 566±297] was lower than those of the control group. The expression level was reduced as the increase of the lymphocyte infiltration focus. ③ The E-cad expression and the p-cat expression was positively correlated in the labial gland of patients with pSS. ④ Howev-er, there was difference in the expression of E -cad and β -cat between patients with positive SSA and negative SSA antibodies. Conclusion In salivary samples, the expression of both E-cad and p-cat in patients with pSS is lower than those of the controls. Anti-SSA/SSB antibodies are important parameters of pSS and they may be involved in the pathogenesis of pSS.

OBJECTIVE To discuss the operative plan for nasal cavity abnormality and evaluate the effect of revision FESS. METHODS A retrospective study of 36 revision FESS performed during 2002.3-2005.5 (age from 17 to 55 years old) is presented. Patients were evaluated with endoscopic examination of the nasal cavities and computed tomography of the sinuses and nasal cavities. Information was also collected during the revision surgery. The plan was designed according to nasal cavity abnormality. RESULTS The reasons of revision FESS: Among 36 cases there are 24 ethmoid sinusitis, 16 cases with recurrence of polyps,13 middle meatal stenosis, 11 middle turbinate adhesion, 10 septal deviation, 5 frontal recess stenosis, 5 sphenoid sisusitis, 3 maxillary ostium obstruction,3 inferior turbinate hypertrophy. Follow up for 6 month～43 month after revision FESS, 22 cases were successful, 10 satisfied and 4 inefficacy. CONCLUSION The primary FESS failure was associated with nasal cavity structure diseases. Meticulous attention in these areas and correcting it during surgery may reduce recurrence and ensure operative effect after revision FESS.

Objective To investigate the apoptosis induction effect of Cantharidin on pancreatic cancer cell line PANC1 and CFPAC-1 and possible mechanism. Methods PANC1 and CFPAC-1 was treated with Cantharidin. Cell growth was determined by MTT. Apoptosis was measured by flow cytometry. Caspase activity was measured by using enzyme chemical method. Apoptosis-related gene expressions were determined by using RT-PCR and Western blotting. Results Cantharidin significantly inhibited the growth of pancreatic cancer cells PANC1, CFPAC-1 and induced apoptosis in a dose-dependent manner. Seventy-two hours after 10 μmol/L Cantharidin treatment, the inhibitory rates of PANC1, CFPAC-1 were (52.95 ± 6.34)％ and (71.21 ±6.30)％. Twenty-four hours after treatment, the early and later period apoptotic cell of PANC1 was increased from 7.35％ to 24.89％, from 6.36％ to 17.73％. The early and later period apoptotic cell of CFPAC was increased from 6.39％ to 24.70％, from 9.21％ to 12.58％ (P＜0.01). Activity of caspase 8 and caspase 9 in PANC1 cells was (155.8 + 11.5)％ and (194.6 ± 14.7)％ when compared with that of control group. Activity of caspase 8 and caspase 9 in CFPAC- 1 was ( 182.5 ± 24.3 ) ％ and ( 215.8 ± 12.2) ％when compared with that of control group ( P ＜ 0. 01 ). The expression of pro-apoptotic genes, TNF-α,TRAILR1, TRAILR2, Bad, Bak and Bid was elevated, the expression of anti-apoptotic Bcl-2 gene was decreased. Conclusions Cantharidin can induce apoptosis in pancreatic cancer cell lines by activating caspase,up-regulating the expression of pro-apoptotic genes and down-regulating the expression of anti-apoptotic genes.

Objective To observe the effect of human immunoglobulin on elderly head and facial post herpetic neuralgia (PHN) and peripheral blood TNF-α.Methods One hundred and twenty-two inpatients with PHN aged ≥65 years old were selected and divided into the observation group (52 cases) and control group (70 cases) by the systematic sampling method.The control group was given the early conventional combined therapy,while on this basis the observation group was intravenously dripped by human immunoglobulin.The incidence rate of PHN and pain visual analog scale(VAS) score at 1,2,3 months after recovery discharge from hospital were recorded in the two groups.Other 20 healthy elderly people were selected as the healthy control group.The TNF-α level was determined in the two patients groups before and after treatment and in the healthy control group.Results The PHN occurrence rate at 1,2,3 months after discharge in the observation group was significantly lower than that in the control group(P＜0.05);the VAS score after treatment and at 1,2,3 months after discharge in the observation group was significantly lower than that in the control group (P＜0.05);serum TNF-α level after treatment in the observation group was significantly lower than that in the control group (P＜0.01),moreover which was close to the level in the healthy control group(P＞0.5).Conclusion Human immunoglobulin can reduce the PHN occurrence in the old people with head and facial herpes zoster and reduce the peripheral blood TNF-α level.

BACKGROUND: Acute lung injury (ALI) is a common and serious complication of severe acute pancreatitis (SAP). The study aimed to investigate the protective effect and mechanism of phosphatidylinositol-3 kinase (PI3K) inhibitor Wortmannin in SAP associated with ALI. METHODS: Ninety rats were randomly divided into three groups: sham operation (SO) group (n=30), SAP group (n=30), and SAP+Wortmannin (SAP+W) group (n=30). SAP model was induced by retrograde injection of 4％ sodium taurocholate into the biliopancreatic duct of rats. The rate of lung water content, myeloperoxidase (MPO), matrix metalloproteinase 9 (MMP-9), protein kinase B (PKB), abdphosphorylation of protein kinase B (P-PKB) activity in the lung tissue were evaluated. RESULTS: In the SAP group, the p-PKB expression in the lung tissue began to rise at 3 hours after modeling, and peaked at 12 hours (P<0.05); the rate of lung water content, MPO and TNF-α activity were also gradually increased, and the degree of lung lesion gradually increased (P<0.05). In the SAP+Wortmannin group, the p-PKB expression in the lung tissue began to rise at 3 hours after modeling, and peaked at 12 hours; it was higher than that in the SO group (P<0.05), but signifi cantly lower than that in the SAP group (P<0.05). The rest indicators in the SAP+Wortmannin group were also signifi cantly decreased as compared with the SAP group (P<0.05). CONCLUSIONS: The expression of phosphatidylinositol-3 kinase/protein kinase B was elevated in severe pancreatitis rats with lung injury. This suggested that PI3K signal transduction pathway is involved in the control and release of proinfl ammatory cytokines TNF-α, which may play an important role in the pathogenesis of severe acute pancreatitis associated with lung injury. This finding indicated that Wortmannin can block the PI3K signal transduction pathway, and inhibit the release of infl ammatory factor TNF-α.

OBJECTIVETo observe the effect of obesity on pharmacokinetics and pharmacodynamics of isoflurane.

METHODSTwenty-six patients undergoing cholecystectomy were divided into obese group (Group A, BMI > or = 27, n = 13) and normal body weight group (Group N, BMI < or = 24, n = 13) according to body mass index (BMI). All patients were given to the same isoflurane anesthesia. Inspired and end-expired concentrations of isoflurane were monitored and uptake fraction of isoflurane were calculated.

RESULTSIsoflurane concentrations of vaporizer in Group A [(1.8 +/- 0.3)%] were evidently higher than those in Group N [(1.5 +/- 0.1)%] at all observed points (P < 0.05 or P < 0.01). Uptake fraction of isoflurane in Group A were higher than those in Group N at observed points (P < 0.05, P < 0.01 or P < 0.001), but there were no differences in the time when isoflurane concentration was lowered to 50% and awake time between the two groups after discontinuing inhaling isoflurane.

CONCLUSIONSObese patients demand higher inspired concentration and uptake of isoflurane than those in normal weight patients but discharge of isoflurane was influenced by obesity within the observed period of (66 +/- 33) min.

Adult ; Aged ; Anesthesia, Inhalation ; Anesthetics, Inhalation ; administration & dosage ; pharmacokinetics ; Body Mass Index ; Cholecystectomy, Laparoscopic ; Dose-Response Relationship, Drug ; Female ; Humans ; Isoflurane ; administration & dosage ; pharmacokinetics ; Male ; Middle Aged ; Obesity ; complications ; metabolism