To investigate the effect of compound Chinese traditional medicine PC-SPES II I in inhibiting proliferation of human prostate cancer cell LNCaP based on the androgen receptor (AR) signaling pathway. The effect of PC-SPES II on LNCaP cell proliferation was detected by MTT assay. According to the findings, at the mass concentration of 180-1 440 mg x L(-1), PC-SPES II significantly inhibited the proliferation of LNCaP cells; the IC50 of PC-SPES II at 24 h and 48 h were 311.48, 199.01 mg x L(-1), respectively. The flow Cytometry detection showed 240 mg x L(-1) PC-SPES II arrested cells in G2/M phase, and an obvious apoptotic peak appeared before G0/G1 peak and rose over time. Meanwhile, Hoechst 33258 staining revealed apoptotic cellular morphology. Annexin V-FITC/PI staining manifested an increase in apoptotic cell ratio at the PC-SPES II concentration of 480 mg x L(-1) in a dose dependent manner. The prostate specific antigen (PSA) secretion of LNCaP cells was tested by PSA ELISA kit. Besides, compared with 25 mg x L(-1) Bic, 480 mg x L(-1) PC-SPES II significantly reduced the cell secretion of PSA. The AR and PSA mRNA and protein expressions were detected by qRT-PCR and Western blot. According to the results, after the induction of LNCaP cells with synthetic androgen 25 μg x L(-1) R1881, 240-480 mg x L(-1) PC-SPES II notably down-regulated the AR and PSA mRNA and protein expressions and inhibited the translocation of AR from cytoplasm to nucleus. In summary, PC-SPES II significantly can inhibit the in vitro proliferation of LNCaP cells and arrest cell cycle arrest in G2/M phase. Its mechanism may be associated with the down-regulation of the AR and PSA expressions and the inhibition of AR nuclear translocation.
Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Male ; Prostate-Specific Antigen ; genetics ; metabolism ; Prostatic Neoplasms ; drug therapy ; genetics ; metabolism ; physiopathology ; Receptors, Androgen ; genetics ; metabolism ; Signal Transduction ; drug effects

OBJECTIVETo investigate the effects of atorvastatin on C-reactive protein (CRP) induced Toll-Like receptor 4 (TLR4)expression on CD14+ monocyte, and the production of proinflammatory cytokines tumor necrosis factor alpha (TNFalpha), interleukin-6 (IL-6), matrix metalloproteinases-9 (MMP-9), and to study the anti-inflammatory mechanisms of statins.

METHODSThe monocytes were isolated from blood of healthy volunteers by the Ficoll density gradient and stimulated by CRP with different doses (5, 25, 50, 100 microg/ml) and different exposure time (6, 12, 24, 48 h). Cells were also incubated with atorvastatin of different doses (1.0, 2.5, 5.0, 7.5, 10.0 micromol/L) in the presence of CRP 50 microg/ml. The protein expression of TLR4 was measured by flow cytometry, mRNA expression of TLR4 and of myeloid differentiation protein (MD2)was detected by quantitative PCR. TNFalpha, IL-6, MMP-9 concentrations in supernatants of cultured medium were measured by ELISA.

RESULTS(1) Compared with the un-stimulated control group, enhanced TLR4 protein expression was already detected at a concentration of 5 microg/ml of CRP and increased in a dose-dependent manner (32.22 +/- 2.80)%, (49.94 +/- 5.58)%, (74.82 +/- 3.24)% and (90.82 +/-2.88)% at 5, 25, 50 and 100 microg/ml CRP. (2) TLR4 protein expression on 50 microg/ml CRP stimulated cells also increased in a time-dependent manner (29.80 +/- 2.70)%, (47.44 +/- 4.41)%, (81.71 +/- 2.92)% and (50.57 +/- 3.34)% after 6 h, 12 h, 24 h, 48 h. (3) When monocytes were incubated with CRP 50 microg/ml and atorvastatin (1.0, 2.5, 5.0, 7.5, 10.0 micromol/L), protein expression [(68.17 +/- 1.71)%, (52.43 +/- 1.38)%, (27.72 +/- 4.55)%, (17.46 +/- 3.20)%, (9.99 +/- 2.81)%] and mRNA expression (82.72%, 67.34%, 48.16%, 30.88%, 13.85%) of TLR4 as well as mRNA expression of MD2 (81.78%, 71.04%, 47.85%, 27.06%, 18.30%) were reduced in a dose-dependent manner. (4) Level of TNFalpha, IL-6 and MMP-9 in supernatants was significantly reduced by atorvastatin (2.5 micromol/L) compared with control group (P < 0.01). When monocyte incubated with CRP 50 microg/ml and atorvastatin 10.0 micromol/L, the level of TNFalpha, IL-6, MMP-9 decreased to (25.8 +/- 2.5) microg/ml, (128.2 +/- 14.7) pg/ml, (65.2 +/- 12.3) ng/ml, respectively.

CONCLUSIONCRP increased the protein expression of TLR4 on CD14+ monocyte in a dose-dependent and time-dependent manner. Atorvastatin can inhibit the signal transduction of TLR4 and reduce proinflammatory cytokines release induced by CRP on CD14 monocyte, and this might be one of the anti-inflammatory mechanisms of atorvastatin.

Anti-Inflammatory Agents ; pharmacology ; Atorvastatin Calcium ; C-Reactive Protein ; metabolism ; Cells, Cultured ; Heptanoic Acids ; pharmacology ; Humans ; Lipopolysaccharide Receptors ; Monocytes ; drug effects ; metabolism ; Pyrroles ; pharmacology ; Toll-Like Receptor 4 ; metabolism

OBJECTIVETo investigate the effects of different anesthesia methods on immune surveillance and tumor metastasis in tumor-bearing rats.

METHODSSeventy-two Fischer 344 rats were randomly assigned into 3 equal groups and anesthetized for 1 h with ketamine (group K), propofol (group P), or neuraxial block (group B). All the rats were subjected to laparotomy followed by intravenous injection of MADB106 tumor cells, and 24 h after the injection, the number and activity of circulating CD3(+), CD4(+), CD8(+), and D4(+)/CD8(+) lymphocyte subsets and NK cellèCD161a(+)éwere assessed. Three weeks later, the lung metastases were counted.

RESULTSCompared with those in group B, the numbers of CD3(+), CD4(+), CD8(+), and CD161a(+) lymphocytes and the activity of circulating NK cells were significantly reduced, and the lung metastases of MADB106 increased significantly in groups K and P (P<0.05 or 0.01 ). The activity of immune surveillance in group K was significantly lower than that in group P except for CD8(+) cells, and the tumor metastases in group K increased significantly in comparison with those in group P (P<0.05 or 0.01).

CONCLUSIONNeuraxial block provides protection of the activity of immune surveillance and reduces tumor metastases in tumor-bearing rats compared with general anesthesia.

Anesthesia, Epidural ; adverse effects ; Anesthesia, General ; adverse effects ; Animals ; Breast Neoplasms ; immunology ; surgery ; Female ; Immunologic Surveillance ; immunology ; Ketamine ; pharmacology ; Lung Neoplasms ; immunology ; secondary ; Male ; Neoplasm Metastasis ; Neoplasm Transplantation ; Nerve Block ; Propofol ; pharmacology ; Random Allocation ; Rats ; Rats, Inbred F344

BACKGROUNDConsiderable evidence suggests that phosphatase of regenerating liver-3 (PRL-3) plays multiple roles in cancer metastasis; however, the molecular mechanisms remain largely unknown. The aim of this study was to identify proteins associated with PRL-3-promoted colon cancer metastasis, by comparative proteomic analysis.

METHODSProteomes of human colon cancer LoVo cells transfected with PRL-3 gene (LoVo-PRL-3) or empty vector PAcGFP-C3 (LoVo-control) were compared using 2D gel electrophoresis. Proteins that varied significantly in concentration were selected and identified using mass spectrometry. Expression of translationally controlled tumor protein (TCTP) mRNA and protein in LoVo-PRL-3 and LoVo-control cells was detected by real-time PCR and Western blotting. Small interfering RNA (siRNA) targeting TCTP was used for silencing TCTP expression in LoVo-PRL-3 cells. Functional significance of TCTP in PRL-3-promoted colon cancer cell proliferation, migration and invasion was investigated by Cell Counting Kit-8 assay and transwell chamber.

RESULTSSeventeen proteins displaying significant and reproducible differences between LoVo-PRL-3 and LoVo-control cells were identified. Ten proteins were upregulated and seven were downregulated in LoVo-PRL-3 cells when compared with LoVo-control cells. Eight identified proteins are associated with distinct steps of tumor metastasis: ubiquitin-like protein ISG15, interleukin-18, TCTP, serpin B5, annexin A3, macrophage-capping protein, ATP-dependent RNA helicase DDX3X, and cathepsin D. Real-time PCR and Western blotting results showed that both TCTP mRNA and protein were significantly increased in LoVo-PRL-3 cells compared to LoVo-control cells. Transfection with TCTP siRNA significantly reduced the expression of both mRNA and protein levels of TCTP in LoVo-PRL-3 cells. Knockdown of TCTP by siRNA inhibited PRL-3-promoted proliferation, migration and invasion of LoVo-PRL-3 cells.

CONCLUSIONOur results imply that TCTP might be a mediator of PRL-3-promoted proliferation, migration and invasion of human colon cancer cells.

Biomarkers, Tumor ; genetics ; metabolism ; Blotting, Western ; Cell Line, Tumor ; Cell Movement ; physiology ; Cell Proliferation ; Colonic Neoplasms ; metabolism ; Humans ; Neoplasm Proteins ; genetics ; metabolism ; Protein Tyrosine Phosphatases ; genetics ; metabolism ; Proteomics ; methods ; Real-Time Polymerase Chain Reaction

【Objective】To assess the difference in volume load between different blood pressure control levels in outpatients with hypertension. To investigate the effect of clinical factors on volume load in outpatients with hypertension. To analyze the clinical baseline characteristics and use of antihypertensive drugs in outpatients with hypertension. 【Methods】 A total of 514 outpatients with hypertension were included from July to November 2017. Clinical indicators including gender，age，height，weight，years of hypertension，blood pressure levels，treatment options for hypertension and comorbidities of these patients were recorded. The body volume load was evaluated by detecting the ratio of extracellular fluid（ECW）to total body water（TBW）using a Bioimpedance Analyzer. Whole body ECW/TBW≥0.39 was defined as high volume load. The effects of clinical factors on the volume load of hypertensive patients and whether there was a difference in volume load between different antihypertensive therapy were analyzed. 【Results】The blood pressure compliance rate of outpatients with hypertension was 15.37% ，which was still very low. Male patients had lower blood pressure compliance rate than female patients. Blood pressure was more difficult to control in patients with older age ，higher body mass index （BMI），higher waist-to-hip ratio（WHR）or longer duration of hypertension. The higher the blood pressure grading，the higher the proportion of combination medication. Diuretics were still the most widely used antihypertensive drugs. Age ，gender and different hypertension grades were the main factors affecting the volume load of hypertensive patients. Volume load was higher in female，older or higher systolic blood pressure（SBP）patients. Among them，age was the most important factor affecting the volume load of hypertensive patients.【Conclusion】The blood pressure compliance rate of outpatients with hypertension was still low. Effectively reducing the volume load was one of the important means to control blood pressure，which was more important in female，older or higher SBP patients.

OBJECTIVETo investigate the protective effects of different concentrations of ketamine against acute lung injury induced by lipopolysaccharide (LPS) in rats and its mechanism.

METHODSForty-eight male Wistar rats were randomized into 4 equal groups, namely the control group, LPS group, ketamine group I (5 mg/kg), and ketamine group II (10 mg/kg). The neutrophil count, protein contents in the bronchoalveolar lavage fluid (BALF) and the wet/dry lung weight ratio were measured 4 h after LPS injection. TNF-alpha, IL-8, NO, iNOS and NF-kappaB were also measured in the lung tissues.

RESULTSIn LPS group, the neutrophil count, protein contents in BALF, the wet/dry lung weight ratio and the levels of tumor necrosis factor-alpha(TNF-alpha), interleukin-8 (IL-8), and NO were all significantly increased compared with the control group (P<0.01). The mRNA expression of iNOS and the protein expression of NF-kappaB were also increased in LPS groups. Ketamine treatment attenuated the increase in wet/dry lung weight ratio, neutrophil count, and protein contents in BALF in a dose-dependent manner. Ketamine also dose-dependently inhibited the production of TNF-alpha, IL-8 , and NO and lowered iNOS mRNA and NF-kappaB protein expression.

CONCLUSIONKetamine can offer protection against LPS-induced acute lung injury in rats by inhibiting the expression of NF-kappaB and attenuating the production of the inflammatory cytokines.

Acute Lung Injury ; chemically induced ; drug therapy ; Animals ; Bronchoalveolar Lavage Fluid ; Interleukin-8 ; metabolism ; Ketamine ; pharmacology ; Lipopolysaccharides ; Lung ; drug effects ; pathology ; Male ; NF-kappa B ; metabolism ; Neutrophils ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; metabolism

Tuberculous encephalopathy (TBE) is an important diagnosis in countries with a high prevalence of tuberculosis. TBE is a life-threatening condition but rarely reported in the modern literature. We reported a case of a man with extensive parenchymal lesions involving the brainstem and right cerebellar hemisphere that resolved after treatment. The clinical, laboratory and pathological features of this case are highlighted and the pathogenesis is discussed.
Aged ; Antitubercular Agents ; therapeutic use ; Brain Diseases ; diagnosis ; drug therapy ; microbiology ; Humans ; Hydrocephalus ; diagnosis ; drug therapy ; microbiology ; Male ; Tuberculosis, Central Nervous System ; diagnosis ; drug therapy ; microbiology

BACKGROUNDOsteopontin (OPN) is one kind of cytokine which can play a number of roles in promoting activation of T lymphocyte, regulating balance between Th1 and Th2, participating in cell-induced immunologic response and stimulating B lymphocyte to express multi-clone antibodies. Some researches have showed that OPN may be involved in the pathogenesis of systemic lupus erythematosus (SLE). The aim of this study was to investigate possible association of a single nucleotide polymorphism (SNP) at position 9250 in exon 7 of the OPN gene (OPN gene 9250) with SLE in Chinese patients.

METHODSTotally 158 patients (18 males and 140 females) fulfilled the revised criteria for SLE by the American College of Rheumatology in 1982 and 180 healthy volunteer controls (34 males and 146 females), all from the south of China, consented to participate in the study. OPN gene 9250 polymorphism was detected by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP).

RESULTSThe frequency of TT genotype of the OPN gene 9250 was significantly lower (52.5% vs 70%, P < 0.05) and the frequency of TC genotype of the OPN gene 9250 was significantly higher (43.7% vs 29.4%, P < 0.05) in SLE patients than in controls. There were significant differences in OPN gene 9250 allele and phenotype frequencies between the SLE patients and controls (P < 0.05). When the SLE patients and controls were separated into men and women, significant differences of frequencies were noted in TT genotype, TC genotype and allele of the OPN gene 9250 in women (P < 0.05) but not in men (P > 0.05).

CONCLUSIONSOPN gene 9250 polymorphism appears to be associated with susceptibility to SLE in Chinese Han ethnic population.

Adolescent ; Adult ; Aged ; China ; ethnology ; Female ; Humans ; Lupus Erythematosus, Systemic ; genetics ; Lupus Nephritis ; genetics ; Male ; Middle Aged ; Osteopontin ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Polymorphism, Restriction Fragment Length

BACKGROUNDThe prevalence of dermatophytoses and the development of new antifungal agents has focused interest on susceptibility tests of dermatophytes. The method used universally for susceptibility tests of dermatophytes was published as document (M38-A) in 2002 by the Clinical and Laboratory Standards Institute (CLSI), dealing with the standardization of susceptibility tests in filamentous fungi, though not including dermatophytes especially. However, it is not a very practical method for the clinical laboratory in routine susceptibility testing. In this test, we developed a novel rapid susceptibility assay-glucose consumption method (GCM) for dermatophytes.

METHODSIn this study, we investigated the antifungal susceptibilities of dermatophytes to itraconazole (ITC), voriconazole (VOC), econazole nitrate (ECN) and terbinafine (TBF) by glucose consumption method (GCM), in comparison to the Clinical and Laboratory Standards Institute (CLSI) M38-A method. Twenty-eight dermatophyte isolates, including Trichophyton rubrum (T. rubrum) (n = 14) and Trichophyton mentagrophytes (T. mentagrophytes) (n = 14), were tested. In the GCM, the minimum inhibitory concentrations (MICs) were determined spectrophotometrically at 490 nm after addition of enzyme substrate color mix. For the CLSI method, the MICs were determined visually.

RESULTSComparison revealed best agreement for TBF against T. mentagrophytes and T. rubrum, since MIC range, MIC50, and MIC90 were identical from two methods. However, for ITC and VOC, GCM showed wider MIC ranges and higher MICs than CLSI methods in most isolates. For ECN against T. rubrum, high MICs were tested by GCM (0.125-16 microg/ml) but not M38-A method (0.5-1 microg/ml). The overall agreements for all isolates between the two methods within one dilution and two dilutions for ITC, VOC, ECN and TBF was 53.6% and 75.0%, 57.1% and 75.0%, 82.1% and 89.3%, and 85.7 and 85.7%, respectively.

CONCLUSIONMeasurement of glucose uptake can predict the susceptibility of T. rubrum and T. mentagrophytes to ECN and TBF.

Antifungal Agents ; pharmacology ; Econazole ; pharmacology ; Glucose ; metabolism ; Itraconazole ; pharmacology ; Microbial Sensitivity Tests ; Naphthalenes ; pharmacology ; Pyrimidines ; pharmacology ; Triazoles ; pharmacology ; Trichophyton ; drug effects ; metabolism ; Voriconazole

OBJECTIVETo investigate the effects of irbesartan for heart protection and on heart nitric oxide (NO) system in diabetic rats.

METHODSThirty adult male Wistar rats were randomly divided into three equal groups, namely control group, diabetes group and irbesartan group. Streptozotocin (STZ, 50 mg/kg) was injected to the abdomen to induce diabetes in the rats. After treatment for 12 weeks, the rats were sacrificed and the urine volume, body weight, ratio of heart to body weight, plasma glucose and glycosylated hemoglobin (HbA1c) were measured. NO levels in the serum and myocardium were determined. Inducible nitric oxide synthase (iNOS) expression was determined by immunohistochemistry, and iNOS mRNA detected by RT-PCR.

RESULTSUrine volume, ratio of heart to body weight, plasma glucose, HbA1C, NO levels in the urine, blood and myocardium in diabetic and irbesartan rats were significantly greater than those of normal controls (P<0.05). The ratio of heart to body weight and NO levels of urine, serum and heart tissue in rats of irbesartan group were significantly decreased as compared with those of diabetes rats (P<0.05). Myocardium iNOS mRNA and protein expression decreased significantly in irbesartan group, but not in diabetes group.

CONCLUSIONSThe abnormality in NO and iNOS mRNA expression might be related to diabetic cardiomyopathy. Irbesartan can decrease iNOS mRNA and protein expressions and reduce NO levels in STZ-induced diabetic rats.

Angiotensin II Type 1 Receptor Blockers ; pharmacology ; Animals ; Biphenyl Compounds ; pharmacology ; Diabetes Mellitus, Experimental ; enzymology ; genetics ; metabolism ; Gene Expression Regulation, Enzymologic ; drug effects ; Immunohistochemistry ; Male ; Myocardium ; enzymology ; metabolism ; Nitric Oxide ; blood ; metabolism ; urine ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Tetrazoles ; pharmacology