This study was aimed to explore the mechanism of Xin-Feng capsule (XFC) on pulmonary function of Sjogren's syndrome (SS) rat model based on the TGF-β1-ERK1 signal pathway. A total of 50 SD rats were ran-domly divided into the normal control (NC) group, model control (MC) group, hydroxychloroquine sulfate (HCQ) group, radix paeoniae alba (total glucosides of paeony capsules, TGP) group, and the XFC treatment group, with 10 rats in each group. Except the NC group, the complete Freund's adjuvant (CFA) and the same rat sub-mandibular gland antigen induced method were applied to each rat after metatarsus injection on two feet and CFA after sufficient emulsification of 0.2 mL submandibular gland protein mixed antigen induced SS. The water drink-ing quantity, body weight change, pulmonary function, ERK1, TGF-β1 protein expression and serum cell factor (IL-17, IL-4) changes were detected in each group. The results showed that compared with the NC group, the body quality and serum IL-4 of MC rats were obviously decreased, and the water drinking quantity, submandibu-lar gland / lung index, submandibular gland pathological score, ERK1, TGF-β1 integral and IL-17 were increased (P < 0.01 or P < 0.05), and the pulmonary function parameter was decreased (P < 0.01 or P < 0.05). Compared with the MC group, the body quality, pulmonary function parameters FEF50 and MMF of the XFC group were in-creased, the IL-4 expression was increased, the water drinking quantity, submandibular gland / lung index, sub-mandibular gland pathological score, serum IL-17 expression, ERK1 and TGF-β1 integral were decreased (P <0.01 or P < 0.05). Compared with the HCQ group, the body quality and IL-17 were significantly decreased in the XFC group (P < 0.01), FEF25, FEF75 and MMF were increased (P < 0.01 or P < 0.05). Compared with the TGP group, the lung index and IL-17 were decreased in the XFC group (P < 0.01 or P < 0.05). It was conclud-ed that the lung function of SS rats was declined, which is closely related to the activation of TGF-β1-ERK1 sig-nal pathway. Traditional Chinese medicine of XFC is able to downregulate the TGF-β1 expression and to restrain epithelium-mesenchymal cell transformation, in order to inhibit ERK1 phosphorylation activation and proliferation, decrease immune inflammation, in order to improve SS and lung function.

In order to explore the method to prepare hypoxia UW solution and the stability and preservation of hypoxia UW solution, UW solution was purged by argon or air for 15 min or 60 at a flow rate of 0.8 or 2 L/min, and the oxygen partial pressure of UW solution was detected. The hypoxia UW solution was exposed to the air or sealed up to preserve by using different methods, and the changes of oxygen partial pressure was tested. The results showed that oxygen partial presure of 50 mL UW solution, purged by argon for 15 min at a flow rate of 2 L/min, was declined from 242+/-6 mmHg to 83+/-10 mmHg. After exposure to the air, oxygen partial pressure of hypoxia UW solution was gradually increased to 160+/-7 mmHg at 48 h. After sealed up by the centrifuge tube and plastic bad filled with argon, oxygen partial pressure of hypoxia UW solution was stable, about 88+/-13 mmHg at 72 h. It was concluded that oxygen of UW solution could be purged by argon efficiently. Sealed up by the centrifuge tube and plastic bag filled with argon, oxygen partial pressure of UW solution could be stabilized.
Adenosine/chemical synthesis ; Allopurinol/chemical synthesis ; Anoxia ; Glutathione/chemical synthesis ; Insulin/chemical synthesis ; Organ Preservation/*methods ; Organ Preservation Solutions/*chemical synthesis ; Oxygen/*analysis ; Partial Pressure ; Raffinose/chemical synthesis

Background Heparanase degrade heparan sulfate side chains of heparan sulfate proteoglycans in the extracellular matrix.Heparanase induces angiogenesis and likely promotes the vascularization of tumor.ObjectiveThe present study is to investigate the expression of heparanase and perlecan in retinas with oxygen-induced retinopathy.Methods Sixty-five clean neonatal C57BL/6J mice were raised in a hyperbaric oxygen box with a volume percentage of 75%±2% for 5 days and then returned to the normal air room.Another 65 matched mice were raised in the normal environment as controls.Evans blue was infused by the superior vena cava in all the mice on postnatal days 12,13,17,21 and 30,afterwards fluorescein angiography was performed and then the mice were sacrificed.The retinas of mice were isolated and prepared and the retinal vessels were examined under a fluorescent microscope and optical microscope.Heranase and perlecan mRNA was detected using reverse transcription PCR (RT-PCR).Heranase and perlecan proteins were detected by Western blot.The analysis of variance was used to compare the mRNA and the protein levels of heranase and perlecan between the experimental and control groups.Results The expression of heparanase mRNA in the retinas of different ages of mice and the different groups showed significant differences (F_(group)=16.303,P=0.000;F_(time)=18.614,P=0.000;F_(interaction)=11.299,P=0.000),and the expression of heparanase mRNA was significantly enhanced in mice from postnatal days 12,13,17 and 21 compared with normal control mice (P=0.001,0.000,0.000,0.001,respectively).The expression of heparanase protein in the retinas of different ages of mice and the different groups followed the same tendency(F_(group)=458.134,P=0.000;F_(time)=78.466,P=0.000;F_(interaction)=71.398,P=0.000).The expression of perlecan mRNA in the retinas of different ages of mice and the different groups showed significant differences (F_(group)=7.351,P=0.013;F_(time)=9.098,P=0.000;F_(interaction)=3.349,P=0.000),and increase in differences also were clearly seen in mice from postnatal days 13,17 and 21 compared with normal control mice (P=0.048,0.000,0.003,respectively).Conclusion The expression of heparanase and perlecan is associated with the development and progression of retinal neovascularization,and perlecan and heparanase together produce a synergistic effect.Heparanase and perlecan may participate in the angiogenesis of oxygen-induced retinopathy.

Objective To observe the changes of lung parameters,the ratios of B and T lymphocyte attenuator(BTLA) and serum cytokines in patients with knee osteoarthritis (KOA),and explore possible molecular mechanism of them.Methods Forty-seven cases of knee osteoarthritis from the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine from 2011 October to 2012 July were analyzed in this study.Pulmonary parameters were detected by spirometer; B and T lymphocyte attenuator(BTLA) was detected by flow cytometry ; Interleukin (IL)-1β,IL-10,matrix metalloproteinase-9 (MMP-9),tissue inhibitor of matrix metalloprotease-1 (TIMP-1) were detected by ELISA;ESR was determined by Westergren method ; hs-CRP was determined by the automatic biochemistry analyzer.Results (1) Compared with NC group,levels of FVC,FEV1,FEV1/FVC,PEF,MEF25.75,MEF50,MEF25,CD3 + BTLA+ T cell,CD4+ BTLA+ Tcell,IL-10,TIMP1 were significantly decreased,IL-1 β,MMP9 were significantly increased.(2)Correlation analysis showed FVC was negatively correlated to Lequesne MG,symptom classify quantization scores,course,MMP9,while positively related to CD3+ BTLA+T cell,IL-10,TIMP1 ;FEV1 was positively correlated to CD3 + BTLA+T cell,CD4+ BTLA+T cell,TIMP1,while negatively correlated to course ; MEF50 was positively related to CD3+BTLA+T cell,CD4+ BTLA+T cell.Conclusion While articular cartilage lesions occurred in KOA patients,the lung function was also declined.The mechanism may be associated with the declination of expression of BTLA,which can cause up-regulating of IL-1 β,MMP9 and down-regulating of IL-10,TIMP1,thus leading to immune dysfunction and abnormal immune response.Those may induce airway injuries and result in lung function decline finally.

Objective To explore the clinical effects of pedicled omentum in preventing anastomotic leakage after resection of colorcctal cancer complicated with intestinal obstruction.Methods The clinicopathologic data and anastomotic leakage rate of 102 patients with colorectal cancer undergoing resection from Dec.2012 to Dec.2015 were analyzed.Results Seven patients in the control group developed anastomotic leakage.Only 1 patient in the experimental group developed anastomotic leakage.The incidence of anastomotic leakage in the control group was 12％,while that in the experimental group was 2％ (x2 =4.250,P =0.039).Of the 7 patients complicating anastomotic leakage in control group,1 died of multiple organ failure,1 was cured with conservative treatment,and 5 were done with diverting stoma.The one leakage in experimental group was cured by conservative treatment.Conclusion Pedicled omentum is useful in the prevention of anastomotic leakage after resection of colorectal cancer in settings of intestinal obstruction.

Objective:To explore the relationship between the polymorphisms in gene FGFR1, FGF10, FGFI8 and the nonsyndromic cleft lip with or without cleft palate (NS CLP) in Chinese population. Methods: Genomic DNA was isolated from peripheral lymphocytes of 75 patients with NS CLP and their parents and 75 unimpaired healthy children. The polymorphisms in FGFRI gene rs13317, p. E467K, p. M3691 and p. S393S, FGF10 gene rs1448037 and FGFI8 gene rs4043716 were detected by applying three-dimensional (3-D) polyacrylamide gel microarray technology. The data were performed using statis-tical analysis : the genotype frequenc+ y and allele frequency between patients with NSCL/P and control subjects were performed. Haplotype relative risk (HRR) , family based association test (FBAT) , and transmission disequilibrium test (TDT) in nuclear family were performed. Results: There were no poly-morphism in FGFR1 gene p. E467K, p. M369I and p. $393S site, the corresponding base was all G. The polymorphisms of rs13317 and rs1448037 were detected and their genotype frequency and allele frequen-cy showed no significant difference between 75 patients with NSCL/P and 75 normal children. TDT, HRR and FBAT were also no significant differences. The genotype frequency of gene FGF18 rs4043716 showed significant difference, but allele frequency were no significant difference. TDT, HRR and FBAT were also no significant difference. Conclusion: Our studies suggest an association between gene FGF18 rs4043716 and the NS CLP in Chinese population, and no association among gene FGFR1 rs13317, p. FA67K, p. M3691, p. S393S and gene FGF10 rs1448037.

OBJECTIVETo observe the anti-inflammatory effect of total saponins of Panax japonicus on LPS-induced RAW264. 7 macrophages.

METHODThe effect of total saponins of P. japonicus of different concentrations on RAW264. 7 cell viability was determined with the MTT method. The NO kit assay was adopted to detect the NO release of total saponins of P. japonicus to LPS-induced RAW264. 7 cells. The enzyme linked immunosorbent assay (ELISA) was used to detect the secretion of tumor necrosis factor-alpha (TNF-alpha) and interleukin 1-beta (IL-1beta). The reverse transeriptase-polymerase chain reaction (RT-PCR) was used to determine the expression of inducible nitric oxide synthase (iNOS) ,TNF-alpha,IL-1beta. The protein expression of nuclear transcription factor-kappaB p65 (NF-kappaB p65) was tested by Western blot.

RESULTThe safe medication range of total saponins of P. japonicus was less than 80 mg x L(-1). Compared with the LPS model group, total saponins of P. japonicus high, middle and low dose groups (0.1, 1, 10, 40 mg x L(-1)) could significantly reduce the secretion of NO, TNF-alpha, IL-1beta of LPS-induced RAW264. 7 cells, and inhibit the expressions of iNOS, TNF-alpha and IL-1beta mRNA and the protein expression of NF-kappaB p65.

CONCLUSIONThis study preliminarily proves the protective effect of total saponins of P. japonicus on LPS-induced RAW264.7 macrophages. Its action mechanism may be related to NF-kappaB signal pathway.

Animals ; Anti-Inflammatory Agents ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Inflammation ; drug therapy ; genetics ; immunology ; Interleukin-1beta ; genetics ; immunology ; Lipopolysaccharides ; adverse effects ; Macrophages ; drug effects ; immunology ; Mice ; NF-kappa B ; genetics ; immunology ; Nitric Oxide ; immunology ; Nitric Oxide Synthase Type II ; genetics ; immunology ; Panax ; chemistry ; Protective Agents ; pharmacology ; Saponins ; pharmacology

AIM: To investigate the signal transduction mechanism of Chlamydia pneumoniae (Cpn) in down-regulating the expression of ATP binding cassette A1 (ABCA1) and ATP binding cassette G1 (ABCG1),the key molecules in cholesterol efflux and atherogenesis,from THP-1-derived macrophages. METHODS: Cpn was propagated in Hep-2 cells. THP-1 monocytes were induced into macrophages by 160 nmol/L phorbol myristate acetate (PMA) for 48 h,and were randomly allocated into 4 groups to incubate continually: control group,50 mg/L low density lipoprotein (LDL); Cpn infection group,Cpn (1×10~6 IFU) and 50 mg/L LDL; Cpn and SP600125 (a special JNK inhibiter) group,THP-1 macrophages were previously treated with different concentrations (1-20 μmol/L) of SP600125 for 1 h,and then infected with Cpn (1×10~6 IFU) and 50 mg/L LDL; SP600125 group,SP600125(20 μmol/L)and 50 mg/L LDL. The expressions of ABCA1/ABCG1 and peroxisome proliferator-activated receptor γ (PPARγ) from each group were detected then. The cholesterol efflux was detected by enzyme-fluorescence. The expressions of ABCA1/ABCG1 and PPARγ mRNA and protein were determined by RT-PCR and Western blotting,respectively. RESULTS: Cpn not only down-regulated the ABCA1/ABCG1 expression,but also down-regulated the expression of PPARγ,which regulated the ABCA1/ABCG1 genes transcriptions. However,the mentioned effects of Cpn infection were restrained by the special JNK inhibitor SP600125 in a dose-dependent manner. CONCLUSION: Chlamydia pneumoniae may down-regulate ABCA1/ABCG1 expression from THP-1-derived macrophages via JNK-PPARγ signal transduction pathway.

Objective:To investigate the effects of Chlamydia pneumoniae(Cpn) on SR-A1 and CD36 expression in THP-1-derived macrophages and role of c-Jun NH_2-terminal signal transduction pathway in the process.Methods:Cpn was propagated in Hep-2 cells.THP-1 monocytes were induced into macrophages by 160 nmol/L phorbol myristate acetate(PMA)for 48h,and were randomly allocated into four groups to be incubated continually: control group;Cpn infection group;Cpn and SP600125(a JNK inhibiter)group and SP600125 group.Lipid droplets in cytoplasm were observed by oil red O staining.The contents of intracellular cholesterol ester were detected by enzyme-fluorescence.The expression of SR-A1 and CD36mRNA and protein were determined by RT-PCR and Western blot, respectively. Results:THP-1-derived macrophages infected with Cpn resulted in large accumulation of lipid droplets and foam cell formation when co-cultured with LDL.Meanwhile,the expression of SR-A1 mRNA and protein were up-regulated by Cpn infection (P<0.05).However,the expressions of CD36 mRNA and protein in THP-1-derived macrophages infected with Cpn were unchanged.Moreover,the up-regulation of SR-A1 and foam cell formation induced by Cpn could be restrained by the JNK inhibiter SP600125 in a dose-dependent manner,and SP600125 had little impact on the expression of CD36 in THP-1-derived macrophages infected with Cpn.Conclusion:The up-regulation of SR-A1 but not CD36 expression is involved in mechanisms of Cpn inducing foam cell formation.And Chlamydia pneumoniae up-regulates the expression of SR-A1 via the JNK signal transduction pathway.This may be a novel mechanism for the foam cell formation induced by Cpn.

Background: Dampness-heat syndrome is a major syndrome type in patients with a sharp deterioration of chronic renal failure (CRF). Qingshen Decoction, a compound traditional Chinese herbal medicine, could relieve the clinical symptoms of CRF patients, and was considered to have a certain reversal effect on rapid deterioration of renal function. Objective: To observe the changes of serum interleukin-8 (IL-8) and IL-18 levels in CRF patients with a sharp deterioration and dampness-heat syndrome, and to explore the curative efficacy of Qingshen Granule. Design, setting, participants and interventions: Sixty CRF patients with a sharp deterioration and dampness-heat syndrome from Department of Nephrology, the First Affiliated Hospital of Anhui College of Traditional Chinese Medicine, were randomly divided into treatment group (30 cases) and control group (30 cases), with another 20 healthy individuals as normal control. The patients in the treatment and control groups were all treated with Jiedu Xiezhuo II, a compound traditional Chinese herbal medicine, given as retention enema. Qingshen Granule was additionally administered to the patients in the treatment group with 1 dosage each time and 3 times a day. The treatment course was one month. Main outcome measures: The levels of serum IL-8 and IL-18 in the normal individuals and before and after treatment in the treatment and control groups were detected. Results: The total response rates of treatment group in clinical efficacy assessment and assessment of syndrome of traditional Chinese medicine (86.67% and 86.67%) were higher than those of the control group (56.67% and 60%), and there were significant differences between the two groups (P<0.05). The levels of serum IL-8 and IL-18 in CRF patients before treatment were obviously higher than those in the healthy individuals (P<0.01), and there were no differences in the levels of IL-8 and IL-18 before treatment between the treatment group and control group (P>0.05). After one-month treatment, the levels of serum IL-8 and IL-18 were markedly decreased in the two groups (P<0.01), and the levels of serum IL-8 and IL-18 in the treatment group were markedly lower than those in the control group (P<0.05). There were differences in decreased degrees of IL-8 and IL-18 levels between the two groups (P<0.05). Conclusion: CRF patients with a sharp deterioration and dampness-heat syndrome have high serum IL-8 and IL-18 levels. Qingshen Granule can reduce the levels of serum IL-8 and IL-18, and improve the renal function and ameliorate the clinical symptoms.