Objective To investigate the differences of glycoprotein non-metastatic melanoma b (Gpnmb) expression between M1 and M2 bone marrow-derived macrophages (BMMφs) in mouse.Meth-ods Primary BMMφs were cultured and then identified by immunofluorescence staining for F 4/80 and flow cytometry testing of CD11b.Interferon-γand lipopolysaccharide were used to induce differentiation of BMMφs towards M1 macrophages and interleukin-4 was adopted to induce differentiation of M 2 macropha-ges.Realtime PCR was performed to analyze mRNA expressions of tumor necrosis factor (TNF-α), induc-ible NO synthase (iNOS), macrophage mannose receptor (MMR), arginase-1 (Arg-1) and Gpnmb.Pro-teins of Gpnmb and MMR were detected by double immunofluorescence staining , Western blot and flow cy-tometry.Results (1) Immunofluorescence staining showed high expression of F 4/80 in BMMφs and flow cytometry results showed that CD11b was expressed in 92.7%±6.1% of BMMφs, suggesting that primary BMMφs were successfully cultured.(2) Compared with M0 BMMφs, mRNAs of TNF-αand iNOS were highly up-regulated in M1 BMMφs (both P<0.01), and mRNAs of MMR and Arg-1 were highly up-regula-ted in M2 BMMφs (both P<0.01), indicating that differentiation of BMMφs towards M1 and M2 BMMφs were successfully induced .(3) Expressions of Gpnmb mRNA and Gpnmb protein were predominantly up-regulated in M2 BMMφs in comparison with those in M0 and M1 BMMφs (both P<0.01).Gpnmb and MMR were co-expressed in M2 BMMφs and 83.2%±9.7% of MMR positive BMMφs expressed Gpnmb. Conclusion Gpnmb expression is significantly increased in M 2 macrophages than that in M 1 macrophages in vitro, indicating that Gpnmb which takes part in the differentiation of macrophages might be used as a marker for identification of M 1 and M2 macrophages .

Objective To observe the expression of glycoprotein non-metastatic melanoma protein B (Gpnmb) in the kidney and urine after ischemic-reperfusion injury (IRI),and explore the relationship between Gpnmb and macrophage phenotypes in the IRI kidney.Methods Male C57BL/6J mice were randomly divided into control group (n =4),sham group (n =4) and IRI group (n =12).Both renal pedieles of mice in IRI group were identified and occluded with microvascular clamps for 30 min.Renal pathological injury was observed by PAS staining.The expression of Gpnmb was examined by real-time PCR and immunofluoresence staining.The location of Gpnmb was observed by flow cytometry and double immunofluoresence staining with F4/80.The mRNA expressions of Gpnmb,CD40,CRR7,CD163 and MMR were examined by real-time PCR.The expression of Gpnmb in the urine was examined by Western blotting and ELISA.Results PAS-stained IRI kidney section showed desquamative epithelia,necrosis debris and a large number of inflammatory cell infiltration.Real-time PCR results showed that there was little expression of Gpnmb in the kidney of control group and sham group.However,the Gpnmb mRNA level in IRI kidneys was highly up-regulated at day 1 and day 2 (both P ＜ 0.01) and followed by a decrease that was similar to the control level at day 3.Double immunofluoresence staining of kidney sections from IRI mice revealed that Gpnmb was predominantly detected in F4/80 positive macrophages.The mRNA expression of Gpnmb was not correlated with M1 macrophage phenotypes CD40 and CCR7,but positively correlated with M2 macrophages phenotypes CD163 and MMR.Western blotting and ELISA result showed that there was significant increase of Gpnmb expression in the urine from IRI mice compared to those of the control group and the sham group (P ＜ 0.01).Conclusions Gpnmb expression is up-regulated in IRI kidney and is associated to M2 macrophages.It may play a role in the process of acute kidney injury.Gpnmb expression is also increased in urine after IR injury and it may be a new biomarker to diagnose AKI.

This study is to investigate the effects of concentration, intestinal section, pH, paracellular route, substrate/inhibitor of enzyme (CYP3A) and proteins (P-gp, MRP2, SGL1) on the absorption of forsythoside A. The absorption of three concentrations (2.6, 5.2, and 10.4 microg x mL(-1)) of forsythoside A in different intestinal segments was studied with phenol red as the marker by rat circulation in situ. The results showed that the residue of forsythoside A with different concentrations had little significant difference from that obtained after perfusing via duodenum, jejunum, ileum and colon, which indicated that the absorption of forsythoside A was passive diffusion and had no difference in different segments of rat intestine. The residue of forsythoside A increased to 466.160 and 463.429 microg respectively when cyclosporine (4 microg x mL(-1)) or midazolam (50 micromol x L(-1)) was added to the circulation fluid, which showed significant difference compared to the control group (P < 0.05). Moreover, the residue of forsythoside A showed a tendency of increase with the increase of cyclosporine or midazolam. When digoxin (50 micromol x L(-1)) or EDTA (10 microg x mL(-1)) was added to the circulation fluid, the residue of forsythoside A decreased to 325.110 and 369.888 microg respectively, which showed significant difference as compared to the control group (P < 0.05). Besides, the residue of forsythoside A showed a tendency of reduction with the increase of digoxin or EDTA. However, there is no significant change in the absorption of forsythoside A when the different concentrations of mannitol were added to the circulation fluid. The results above indicated that the absorption of forsythoside A was mainly passive diffusion and involved paracellular route at the same time. In addition, the substrates of P-gp or CYP3A had dose-dependent effect on the absorption of forsythoside A.

Objective To clarify the diagnostic value of salivary gland ultrasonography in Sj(o)gren's syndrome (SS),and its correlation with the disease activity index and important organs involvement were analyzed.Methods A total of 116 patients with SS were involved,including 71 cases of primary SS and 45cases of secondary SS.Ultrasonography examination of major salivary glands was conducted for these patients,at the same time the clinical data including inflammatory parameters,the immunological parameter and the involved systems were collected.Ultrasonography examination was conducted in 49 cases as the control group.Use t test,x2test and analysis of variance for statistical analysis.Results The positive rate of salivary gland uhrasonography in SS (56/116,48.3％) was significantly higher than that of the normal control groups (1/49,2.0％),(The chi-square value was 32.57,P＜0.05),the sensitivity of salivary gland ultrasonography in primary SS (62.0％) was obviously higher than secondary SS (27％),(The Chi-square value was 13.75,P＜0.01).The specificity of salivary gland ultrasonography was 98％.The scores of salivary gland ultrasonography had shown positive correlation with the erythrocyte sedimentation rate,the levels of Immunoglobulin (Ig)G,and RF(r=0.234,0.353,0.176;P=0.002,0.000,0.013),and negative correlation with the white blood cell count (r=-0.292,P=0.000).Conclusion Salivary gland ultra-sonography provides additional evidence for the diagnosis of SS,particularly in primary SS groups.The scores of ultrasonography are correlated with inflammatory biomarkers,indicating that salivary gland ultrasonography is related to disease activity.

T helper (Th) 17 cells are a kind of Th cell subset, and are distinct from the Th1 and Th2 cells and produce interleukin-17A (IL-17A, IL-17). Th17 cells have a mechanism of independent differentiation and developmental regulation. The differentiation and cytokine secretion of Th17 cells are regulated by TGF-β, IL-6, IL-23 and orphan nuclear receptor (RORγt). IL-17A induces pro-inflammatory cytokines and chemokines, mediating neutrophil recruitment. Increasing evidence implicated involvement of Th17 cells in anti-glomerular basement membrane disease, lupus nephritis and pauciimmune glomerulonephritis. In this review, we discussed the discovery of Th17 subset, its properties, its relationship with other Th subsets and involvement of Th17 cells in glomerulonephritis.
Animals ; Glomerulonephritis ; immunology ; Humans ; Interleukin-17 ; metabolism ; physiology ; Interleukin-23 Subunit p19 ; physiology ; Interleukin-6 ; physiology ; Nuclear Receptor Subfamily 1, Group F, Member 3 ; physiology ; Th17 Cells ; immunology ; metabolism ; Transforming Growth Factor beta ; physiology

Objective To investigate the correlation of infiltration of mast cells in kidney with renal interstitial fibrosis, expression of TGF-β1 and stem eel] factor (SCF) in rat models withprotein-overload nephropathy. Methods Sixty uninephrectomized SD rats were randomly divided into model group [intraperitoneal injections of bovine serum albumin (BSA)] and control group (intraperitoneal injections of equal volume of saline). Ten rats from both groups were sacrificed respectively at week 3, 7 and 11 after injection. 24 h urinary protein and serum biochemistry of these SD rats at the time of sacrifice were measured. The intensity of mast cell infiltration was examined by toluidine blue (TB) staining and immunohistochemistry using a monoclonal anti-MC chymase antibody. The expression of TGF-β1 and SCF was detected byimmunohistochemistry, using a monoclonal mouse anti-rat TGF-β1 antibody and a polyclonal rabbstanti-rat SCF antibody. Results Severe proteinuria was induced in the rats by BSA injectionpeaked at week 7 [(199.1±98.4) mg/d] after the BSA injection and gradually decreased until week11 [(133.7±67.8) mg/d]. Renal injury was accompanied with chymase-postitive and TB-postitive mast cell infiltration, in close proximity to areas of interstitial fibrosis. With aggravation oflesions degree, the number of mast cells increased,the difference between the modal rats and control rats was significant (P<0.05). Immunostainahle expression of SCIF and TGF-β1 was detected in tubular as well as interstitial cells, and increased with the BSA injection. The difference between the model rats and control rats was significant (P<0.05). Mast cells were positively correlated with interstitial fibrosis (r=0.772, P<0.01), expression of TGF-β1 (r=0.521, P<0.01) and SCF(r=0.916,P<0.01). Conclusions Increased infiltration of mast cells is involved in interstitial fibrosis of rats with protein-overload nephropathy. Proteinuria may attract mast cells to kidney by chemot actions of SCF,and mast cells may contribute to the development of renal fibrosis by secreting chymase and increasing expression of TGF-β1.

OBJECTIVETo study the intestinal absorption kinetics characteristic of the main four active ingredients in Shuanghuanglian oral liquid (SHL) and to investigate the influence of herbal compatibility in SHL on absorption of main effective ingredients.

METHODThe in situ rat circulation model was used to investigate the concentration change differences of the four active components in SHL during perfusion.

RESULTThe absorption quantity of different concentrations of baicalin, chlorogenic acid, phillyrin and forsythoside A ranging from 40-160, 6-24, 3-12, 2.6-10.4 mg x L(-1) respectively was linear with concentration and showed no saturation at high concentration. The absorption rate constant K(a) and the hourly absorption percentages A were essentially unchanged. The pH changing from 5.0-7.43 had little influence on the absorption of phillyrin except baicalin, chlorogenic acid and forsythoside A. The calculated K(a) and A of the four active ingredients had no significant differences from that obtained after perfusing via duodenum, jejunum, ileum and colon; The calculated K(a) and A of baicalin in Scutellariae Radix (SR), chlorogenic acid in Lonicerae Japonicae Flos (LJF) and phillyrin in Forsythiae Fructus (FF) had no significant differences compared with that in SHL, but the calculated K(a) and A of forsythoside A in FF were obviously superior to that in SHL.

CONCLUSIONThe intestinal absorption of the four active ingredients in SHL was mainly passive diffusion and had no difference in different segments of rat intestine; the compatibility of SHL compounds changed the absorption of forsythoside A in FF obviously.

Administration, Oral ; Animals ; Chlorogenic Acid ; analysis ; pharmacokinetics ; Drugs, Chinese Herbal ; chemistry ; Flavonoids ; analysis ; pharmacokinetics ; Glucosides ; analysis ; pharmacokinetics ; Glycosides ; analysis ; pharmacokinetics ; Hydrogen-Ion Concentration ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo research the influence of glycyrrhiza extract on the pharmacokinetics characteristic parameters of daphnetin, which was aimed to explore the rationality of concert application of drugs.

METHODThe rats received intragastric administration of daphnetin and glycyrrhiza extract containing the same daphnetin respectively. The blood concentration of daphnetin was assayed by LC-MS. The data was processed by program DAS2.1.1.

RESULTGlycyrrhiza extract can reduce the t(1/2), tmax and Ke of daphnetin, while increased the Ka and AUC(0-infinity).

CONCLUSIONGlycyrrhiza extract promoted the oral absorption of daphnetin, slowed down the elimination and increased the biological availability.

Animals ; Drug Interactions ; Drugs, Chinese Herbal ; pharmacology ; Glycyrrhiza ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tissue Distribution ; drug effects ; Umbelliferones ; pharmacokinetics

Objective To investigate the prevalence of chronic kidney disease (CKD) and associated factors in Changsha county of Hunan province. Methods Using a stratified, multistage sampling, 1950 residents (older than 20 years old) from 3 towns of Changsha county were randomly selected to be interviewed and tested for the kidney damage indicators and the associated factors with CKD. Results Eligible data of 1727 subjects were enrolled in the study. After the adjustment of age and gender compenent, the prevalence of albuminuria was 8.5%, hematuria 5.1%, and reduced eGFR 1.5%. Approximately 14.6% subjects had at least one indicator of kidney damage, and the awareness rate was 16.5%. Age, hypercholesteremia, hypertriglyceridemia, hypertension and diabetes were independently correlated with albuminuria. Female, age, hypertriglyceridemia and hyperuricemia were independently correlated with reduced renal function. Female was independently correlated with hematuria. Conclusions The prevalence of chronic kidney disease is 14.6% and the awareness rate is 16.5% in suburban adult population of the central south area of China. The spectrum and correlated factors of CKD in this county undergoing fast economic development are close to those of Guangzhou and developed countries.

Breast cancer is the most common malignant tumor in women. The key to the prognosis of breast cancer lies in early diagnosis and early treatment.At present, the early diagnosis of breast cancer mainly relies on imaging methods, and there has been a lack of clinical methods for early diagnosis of it.Exosome are small molecules selectively released from breast cancer cells to the peripheral circulation for a certain period of time. miRNA contained in them are short, non-coding regulatory RNA, which can bind to the 3′UTR region of target genes and regulate the post-transcriptional expression of target genes. Exogenous miRNA can affect the development of breast cancer, It can provide a variety of information indicating the presence of breast cancer cells, clinical staging, molecular typing and other information, which may be put into clinical application in the future as an ideal marker for the early diagnosis of breast cancer.This article reviews the research and application of exogenous miRNA in early diagnosis of breast cancer.