Background T helper cell 17 (Th17),a newly discovered subset of CD4+ T cells,have been found to play an important role in dry eye disease in animal model.Further investigation should be done on the immunopathogenesis of Th17 cells in dry eye patients.Objective This study was to analyze the expression status of interleukin-17 (IL-17) and IL-22 in tear and supernatant of peripheral blood mononuclear cells (PBMCs) of dry eye patients and their correlation with clinical symptom and sign.Methods Twenty Sj(o)gren syndrome (SS)patients,twenty non-Sj(o)gren syndrome (NSS) patients were included in Wuxi Second Hospital from 2010 to 2011,and twenty healthy volunteers were recruited at the same period.All of subjects understood the purpose and procedure of research and written informed consent was obtained form each subject initial of this study.Dry eye symptom questionnairs were self-answered and multiple dry eye disease-related clinical tests,including the breakup time of tear film (BUT),Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescein staining were performed.The periphery blood of 3 ml and tear were collected in all the subjects,and IL-17 and IL-22 levels in supernatant of PBMCs and tear were detected by enzyme-linked immunosorbent assay (ELISA).The correlations between levels of IL-17 and IL-22 with BUT,S Ⅰ t,corneal fluorescein staining and dry eye scores were analyzed.Results The dry eye scores reduced,BUT prolonged,S Ⅰ t increased and corneal fluorescein dye decreased from SS group,NSS group to normal control group,with significant differences among the three groups (dry eye scores:H =40.81,P＜0.01 ; BUT:H =40.15,P＜0.01 ;S Ⅰ t..H=50.07,P＜0.01 ;corneal dye scores:H=40.52,P＜0.01).The concentration of IL-17 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (964.92±124.83)ng/L,(718.85± 115.89)ng/L and (341.95±85.08) ng/L,showing a statistically significant difference among them (F=162.95,P＜0.01).The levels of IL-17 in the tear were (440.69±126.09) ng/L,(364.33±126.85) ng/L and (61.16±11.60) ng/L in the SS group,NSS group and normal control group respectively,exhibiting an elevated level in the SS group and NSS group compared with the control group (F=75.27,P＜0.01).In addition,the levels of IL-22 in the supernatant of PBMCs in the SS patients,NSS patients and normal controls were (98.77± 11.27) ng/L,(79.65 ± 11.01) ng/L and (32.78±9.34) ng/L,and those in the tear were (22.22 ± 8.96) ng/L,(14.92 ±4.35) ng/L and (10.47 ± 2.67) ng/L,with significant differences among the three groups (F =206.27,P＜0.01 ;F =19.87,P＜0.01).The significant correlations were found between the IL-17 and IL-22 concentration in the supernatant of PBMCs and tear with corneal fluorescein staining scores and S Ⅰ t.Conclusions The contents of IL-17 and IL-22 in PBMCs and tear upregulate in the SS and NSS patients,indicating that Th17 plays a key role in the immunity mechanism of dry eye.

Background Mechanism of dry eye disease has not been established clearly.Some increasing evidences showed that inflammation plays an important role in pathogenesis and development of dry eye disease.However,the association of interleukin-6 (IL-6) with dry eye has not been clarified.Objective This study was to assess the correlation of IL-6 levels in tear and serum of patient with dry eye disease and clinical symptom.Methods Twenty patients with Sj（o）gren syndrome (SS) and 20 cases with non-Sj（o）gren syndrome (NSS) were enrolled from Second Affiliated Hospital of Nanjing Medical University,and 20 healthy volunteers were recruited as the normal controls.Symptom questionnaires were self-answered and multiple dry eye-related clinical tests were performed,including tear film breakup time (BUT),Schirmer Ⅰ test (S Ⅰ t) and corneal fluorescence staining.IL-6 concentrations in serum and tear were detected by enzyme-linked immunosorbent assay (ELASA).The correlation between IL-6 level and tear film and ocular surface parameters was analyzed by Spearman rank correlation.Written informed consent was obtained from each subject initial of this trial.Results The scores of symptom questionnaires were 18.50 (11.75,23.00),23.00 (19.00,32.00) and 2.00 (0.25,4.00) in the SS group,NSS group and the normal control group,and significantly increasing values were seen in the SS group and NSS group compared with the normal control group (P=0.00,0.00).BUT values in the SS group and NSS group were 2.50 (2.00,3.88) seconds and 3.50 (2.13,4.00) seconds and were lower than 15.00 (13.13,17.00) seconds in the normal control group (P=0.00,0.00).S Ⅰ t values in the SS group,NSS group and the normal control group were 2.00 (1.50,2.88),4.00 (3.00,5.75) and 19.00 (18.00,25.00)mm,showing significant differences between the SS group and the normal control group (P =0.00),the SS group and the NSS group (P =0.00) or the NSS group and the normal control group (P=0.00).The marked elevation of keratoepithelioplasty score was seen between the SS group and the normal control group (5.75 (4.00,7.75) vs.0.00 (0.00,0.75) (P =0.00) or the NSS group and the normal control group (4.50(3.63,6.00) vs.0.00(0.00,0.75)) (P=0.00).Concentrations of IL-6 in serum and tear in the SS group,NSS group were (131.47±21.04) μg/L and (77.81 ± 15.68) μg/L and were significantly higher than (31.62±8.57) μg/L of the normal control group,and those in the tear were (33.44±8.01) μg/L,(18.78 ±5.73) μg/L and (8.77 ± 3.43)μg/L,respectively in the three groups,with a significant reduce in the normal control group (all P=0.00).tL-6 levels in serum and tear were positively correlated with the score of symptom questionnaires or keratoepithelioplasty and negatively correlated with BUT and S Ⅰ t (all P =0.00).Conclusions IL-6 levels in the tear and serum of the dry eye patient are associated with the severity of the disease.IL-6 levels in serum and tear may be objective,sensitive and reliable indicators of dry eye.

This study aims to investigate the virological impact of the stalk region and cysteine (C) in neuraminidase (NA) of influenza A/Anhui/1/05 (H5N1) and A/Ohio/07/2009 (H1N1) viruses. The NA of A/ Anhui/1/05 (H5N1), defined as AH N1, lacked 20 amino acids (including C, defined as s20) as compared with NA of A/Ohio/07/2009 (H1N1) (defined as 09N1). We deleted s20 of 09N1 to construct 09N1-s20, and inserted s20 into AH N1 to construct AH N1+s20. To investigate the impact of C on the biological function of NA, we deleted C in 09N1 to construct 09N1-C and inserted C into AH N1 to construct AH N1-C. The pseudo-type viral particle (pp) system was used to evaluate the impact of these mutants on virology. The combination of 09N1-C and 09H1 (defined as 09H1::09N1-C) showed an infectivity 8 times that of the wild type 09H1::09N1, while the infectivity of the combination of AH N1+C and AH H5 (defined as AH H5::AH N1+C) was much lower than that of the wild type AH H5::AH N1. The infectivity of the combination of 09N1-s20 and 09H1 (defined as 09H1::09N1-s20) was 4 times that of the wild type 09H1::09N1; the infectivity of the combination of AH N1+s20 and AH H5 (defined as AH H5:: AH N1+s20) was 1/7 that of the wild type AH H5::AH N1. The co-existence of 09N1-C and AH H5 displayed 6 times the infectivity of AH H5::09N1, while the infectivity of 09H1::AH N1+C was very low. Multimer analysis showed that in the wild type 09N1, the forms of NA were dimer > tetramer > monomer; the major component of NA in 09N1-C was monomer; in 09N1-s20, the forms of NA were monomer > dimer. AH N1 was mainly composed of monomer; in AH N1+s20, the forms of NA were dimer > monomer > tetramer; in AH N1+C, the forms of NA were dimer > tetramer. Deletion of C or s20 from 09N1 did not change the expression of NA. The study suggested that deletion of C from the stalk region of NA in A/Ohio/07/2009 (H1N1) increases infectivity. Insertion of C into NA's stalk region of A/ Anhui/1/05 (H5N1) significantly decreases infectivity. Cysteine deletion in the stalk region is important for the infectivity of A/Anhui/1/05 (H5N1) and A/Ohio/07/2009 (H1N1). It may interfere with the infectivity via changes in NA polymerization.
Amino Acid Motifs ; Humans ; Influenza A Virus, H1N1 Subtype ; chemistry ; enzymology ; genetics ; pathogenicity ; Influenza A Virus, H5N1 Subtype ; chemistry ; enzymology ; genetics ; pathogenicity ; Influenza, Human ; virology ; Neuraminidase ; chemistry ; genetics ; metabolism ; Viral Proteins ; chemistry ; genetics ; metabolism ; Virulence

In this study, SD rats were orally administrated with oteracil potassium (300 mg . kg-1 . d-1 ) to prepare the hyperuricemia model, and divided into normal, model, Allopurinol, LE high dosage, middle dosage and low dose (200, 100, 50 mg . kg-1 . d-1) groups. The rats were orally administrated with test drugs 1 hour later after being orally administrated with Oteracil potassium. After 7 days, serum uric acid, serum creatinine, uric acid and expression of relevant transporters in kidney were tested to study the regulatory effect of leonurus extracts on serum uric acid, renal function and relevant transporters in kidney of rats with hyperuricemia. Compared with the model group, the leonurus extract group could significantly down-regulate serum uric acid and creatinine levels of rats with hyperuricemia, and increase the urine uric acid level. Meanwhile, leonurus extracts could notably down-regulate the mRNA expressions of urate transporter 1 (URAT1) and glucose transporter 9 (GLUT9), up-regulate the mRNA expressions of organic cation transportanter (OCT) and Carnitine transporter (OCTN) and promote the excretion of uric acid of kidney.
Allopurinol ; pharmacology ; Animals ; Blood Urea Nitrogen ; Creatinine ; blood ; Disease Models, Animal ; Down-Regulation ; Gene Expression Regulation ; drug effects ; Hyperuricemia ; blood ; drug therapy ; Kidney ; drug effects ; Leonurus ; chemistry ; Male ; Organic Anion Transporters ; genetics ; Oxonic Acid ; administration & dosage ; Plant Extracts ; isolation & purification ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Specific Pathogen-Free Organisms ; Up-Regulation ; Uric Acid ; blood

OBJECTIVETo investigate the preventive effects of a compound Danshen preparation (DSC) on long-term gastric lipid emulsion administration-induced nonalcoholic steatohepatitis in rats.

METHODSTwenty-seven 3-month-old SD rats were randomized equally into 3 groups and subjected to daily intragastric administration for 20 weeks of distilled water (control), lipid emulsion at 5 ml/kg (model group), and lipid emulsion plus DSC at 5.0 g/kg (DSC treatment group). After blood glucose (BG) determination, the rats were sacrificed for measurement of serum TC, TG, HDL-c, AST, and ALT, and the liver was weighed and pathologically examined.

RESULTSCompared with the control group, the rats in the model group showed significantly increased BG, TC, LDL-c, arteriosclerosis index (AI), AST, ALT, liver weight, and liver index (P<0.01) and decreased HDL-c (P<0.01), while TG remained unchanged. Fatty degeneration, hydropic degeneration and necrosis with inflammatory cell infiltration were observed in the liver of the rats in the model group. Compared with the model group, the rats in DSC groups showed decreased BG, AI (P<0.01), liver weight, liver index, AST, and ALT (P<0.05) and increased HDL-c, with milder pathological changes in the liver.

CONCLUSIONSLong-term gastric perfusion of lipid emulsion causes lipid metabolic disorder and nonalcoholic fatty liver disease in rats characterized by increased TC and decreased HDL-c. DSC can significantly increase HDL-c and provide partial protection of the liver against the damages by the lipid emulsion.

Animals ; Drug Administration Routes ; Drugs, Chinese Herbal ; therapeutic use ; Emulsions ; Fatty Liver ; chemically induced ; prevention & control ; Female ; Lipids ; administration & dosage ; toxicity ; Male ; Phenanthrolines ; therapeutic use ; Phytotherapy ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry ; Time Factors

OBJECTIVETo observe the effects of Danshen root compound (DSC) on blood lipid and bone biomechanics in mice with hyperlipemia-induced osteoporosis.

METHODSForty Kunming mice were randomized into 5 equal groups, and were given intragastric administration with distilled water (control), lipid emulsion (LE) at the daily dose of 5 ml/kg, LE plus simvastatin, LE plus DSC at 5.0 g/kg (DSC-L group), and LE plus DSC at 10.0 g/kg (DSC-H group), respectively. Serum TC, TG, and HDL-c levels and left femur hydroxyproline, calcium and phosphate contents were measured in the rats, with the right femur taken for bone biomechanical test.

RESULTSCompared with those in the control group, serum TC, LDL-c and AI of the mice increased and HDL-c, Hyp and bone calcium decreased significantly (P<0.01) with lowered bone biomechanical properties. Compared with those of the LE model group, AI decreased and HDL-c increased significantly in DSC-L and DSC-H groups (P<0.01), and the bone biomechanics in DSC-H group was improved.

CONCLUSIONLong-term intragastric administration of lipid emulsion causes lipid metabolic disorder and induces osteoporosis due to hyperlipemia in mice. DSC can significantly increase HDL-c and partially prevent the occurrence of osteoporosis in mice.

Animals ; Biomechanical Phenomena ; Bone and Bones ; drug effects ; metabolism ; physiopathology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Hyperlipidemias ; complications ; drug therapy ; Lipids ; blood ; Male ; Mice ; Osteoporosis ; blood ; etiology ; prevention & control ; Phenanthrolines ; pharmacology ; therapeutic use ; Phytotherapy ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo study the effect of dimethoate on the expression of heat shock protein 70 (HSP70) in peripheral blood lymphocytes of human beings and to explore the feasibility of HSP70 in biomonitoring among workers exposed to organophosphorous pesticides.

METHODSPeripheral blood lymphocytes were isolated from subjects, comprising 11 people of the control group and 35 workers of the exposure group exposed to dimethoate. Flow cytometry was used for detecting both the basic level and the level of the dimethoate-induced expression of HSP70. The activity of whole blood acetylcholinesterase (AChE) was examined at the same time. Then the potential influential factors to HSP70 expression and AChE activity were analyzed.

RESULTSThe basic level of HSP70 expression in the exposure group and the control group was 41.24% +/- 10.45% and 23.97% +/- 4.29% respectively. The activity of AChE in these two groups were (125.23 +/- 7.97) and (145.36 +/- 8.78) U/ml respectively. Both differences were statistically significant (P < 0.01). Among the exposure group, the basic level of HSP70 expression of the two categories comprising operators and packers, were 47.34% +/- 11.87% and 38.05% +/- 8.20% respectively (P < 0.05), while there was no significant difference (P > 0.05) in AChE activity between these two categories. The factors that had significant influence on the HSP70 basic level of the exposure group were the health condition, the environmental concentration of dimethoate and the exposure time in order, according to their significance of influence. At least 88% variance of HSP70 could be explained by these factors. The only factor that could influence AChE activity significantly was the exposure time, and it could only explain about 12% variance of AChE activity. After the treatment of dimethoate in vitro, the level of the induced expression of HSP70 in the control group was significantly higher than that of the exposure group (P < 0.01). The increasing order was the control group, the group of packers and the group of operators according to the increasing extent and there were significant difference among them (P < 0.01). The factors that could significantly influence the change ratio of HSP70 expression were the environmental concentration and the exposure time.

CONCLUSIONHSP70 is a potential index that can reflect the individual and environmental conditions of workers exposed to dimethoate comprehensively.

Acetylcholinesterase ; blood ; Adult ; Cells, Cultured ; Dimethoate ; toxicity ; Female ; HSP70 Heat-Shock Proteins ; biosynthesis ; Humans ; Insecticides ; toxicity ; Lymphocytes ; drug effects ; metabolism ; Male ; Middle Aged ; Occupational Exposure

Objective To study the sources and the relationship between the management and the outcome of hemorrhage after cephalic pancreatoduodenectomy.Methods The clinical data of 370 patients who underwent pancreatic resection at the Lihuili Hospital and the Second Affiliated Hospital of Zhejiang University were retrospectively analyzed.Results Postoperative bleeding occurred in 35 patients with 11 deaths.Among those intraabominal bleeding occurred in 14 cases and gastrointestinal hemorrhage occurred in 22,with one case suffering from both.Bleediug developing within 72 hours after operation in 12 cases (early-stage group),which was caused by improper intraoperative homeostasis.In other 23 cases,bleeding 72 hours after operation(later stage group)was caused by the erosion following pancreatic and/or bile leakage.Relaparotomy was performed in 13 cases and endoscopic homeostasis was performed in 3. Relaparotomy or endoscopic homeostasis was superior to that of conservative therapy in the early-stage group (P0.05).Pancreatic or bile leakage was identified as the significant risk factors for the postoperative bleeding.Conclusions In order to prevent the postoperative hemorrhage and to reduce the mortality of pancreatic resection,skillful techniques,expeditious homeostasis,proper management of stump pancreas and the prevention of pancreatic and bile leakage are essential.

OBJECTIVETo investigate the association between the expression of galectin- 3 protein and peritoneal metastasis in gastric cancer.

METHODThe expressions of galectin- 3 was detected in matching- samples including primary gastric cancer lesions,lymph node metastases,peritoneal metastases and paratumor normal tissues by immunohistochemistry. All specimens were gained from 35 patients who had synchronous peritoneal metastasis from gastric cancer.

RESULTSThe over- expression of galectin- 3 was observed in 97% (34/35) of the gastric cancer lesions, the peritoneal metastases and the lymph node metastases,whereas in 14% (5/35) of paratumor normal tissues. There were significant differences in the expression of galectin- 3 between paratumor normal tissues and the gastric carcinoma lesions,peritoneal metastases and lymph node metastases (P< 0.05),but there were no significant differences among the gastric cancer lesions,the peritoneal metastases,and the lymph node metastases (P> 0.05).

CONCLUSIONThe expression of galectin- 3 in gastric cancer lesions can be used as a biological marker of peritoneal metastasis from gastric cancer before operation and as a prognostic factor of gastric cancer.

Galectin 3 ; metabolism ; Humans ; Immunohistochemistry ; Lymph Nodes ; metabolism ; pathology ; Lymphatic Metastasis ; Neoplasm Staging ; Peritoneal Neoplasms ; metabolism ; pathology ; secondary ; Stomach Neoplasms ; metabolism ; pathology

Objective: To explore the distribution features of microvolt T-wave alternation(MTWA) through exercise stress test (EST) in coronary artery disease (CAD) patients with MTWA changes after revascularization. Methods: MTWA was measured in pre-cordial ECG leads in 326 patients with suspected CAD. Based on coronary angiography and coronary CTA, the patients were divided into 4 groups: ① Control group, patients without coronary stenosis, n=101, ② Coronary stenosis<50％ group, n=99, ③ 50％ ≤ Coronary stenosis<70％ group, n=53 and ④ Coronary stenosis ≥ 70％ group, n=73; MTWA was compared among different groups. 95 patients with coronary stenosis ≥ 50％were further divided into 2 subgroups: R (right coronary)stenosis ≥ 50％ subgroup, n=23 and LAD (left anterior descending branch) stenosis ≥ 50％ subgroup, n=72; MTWA was respectively compared to Control group. In addition, MTWA was collected from 103 patients with percutaneous coronary intervention (PCI) as PCI group, MTWA was compared to Coronary stenosis ≥ 70％ group. Results: MTWA was obviously higher in Coronary stenosis ≥ 70％ group than the other 3 groups, all P<0.01. Compared with Control group, Rstenosis ≥ 50％ subgroup had increased MTWA in V4-V6 pre-cordial leads, P<0.05; LAD stenosis≥50％ subgroup had increased MTWA in V1-V2 pre-cordial leads, P<0.01. Compared with Coronary stenosis ≥ 70％group, PCI group showed reduced MTWA, P<0.01. Conclusion: CAD patients with severe coronary stenosis (≥70％) had increased MTWA; MTWA distribution in body surface was approximately corresponding to coronary stenosis site and PCI may decrease MTWA in CAD patients.