Objective To develop a multiplex PCR-based reverse line blot(mPCR/RLB) hybridization assay to detect,simultaneously,seven genes encoding AR-erm(A/TR),erm(B),mef(A/ E),tet(M),tet(O),aphA-3,aad-6 and two AR-related genes,int-Tn and mreA in group B streptococcus.Methods Nine pairs of specific primers and Oligonucleotide probes targeting erm(A/TR), erm(B),mef(A/E),tet(M),tet(O),aphA-3,aad-6 int-Tn and mreA respectively were modified according to former studies or designed in this study.The primers and probes were labeled with biotin and amino,respectively.The nine genes were amplified simultaneously in the same tube.PCR product hybridized with the probes labeled in the BiodyneC nylon membrane to detect the nine genes.To detect the sensitivity and specificity of the method developed,PCR with single pair of primer targeting each gene were tested in 318 isolates tested and the results were compared with the one abtained by RLB.Results The nine resistance-related genes could be successfully detected by mPCR/RLB assay developed in this study.Based on sequencing,21 of 22 isolates with mef had mef(E)and eight of 353 with int-Tn had an atypical sequence.Except for the above 29 genes,all the others corresponded well with the results obtained by single pair primer PCR.Conclusion The mPCR/RLB assay developed in this study is simple,rapid and suitable for surveillance of antibiotic resistance in GBS.

As one of the most serious types of psoriasis, pathogenesis of erythrodermic psoriasis (EP) is unclear so far. In this study, we aimed to detect the levels of Th1/Th2 cytokine-associated transcription factors and T-lymphocyte clone in peripheral blood mononuclear cells (PBMCs) derived from EP patients, and gene expression level of T-bet/GATA-3 in skin lesion. The potential role of Th1/Th2 reaction pattern played in the pathogenesis of EP was also discussed. Serum levels of IFN-γ, IL-2, IL-4 and IL-10 were quantified by ELISA among 16 EP patients, 20 psoriasis vulgaris (PV) patients and 15 healthy controls. The expression levels of T-bet/GATA-3 in the skin lesion and PBMCs were examined by real-time qPCR. The ratio of Th1/Th2 was measured by flow cytometry. The levels of IFN-γ, IL-2, IL-4 and IL-10 were higher in EP patients than in the healthy controls. The levels of IL-4 and IL-10 were 69.44±11.45 and 12.62±4.57 pg/mL, respectively, in EP patients, significantly higher than those in PV patients and healthy controls (P<0.05). Flow cytometry revealed the levels of both Th1 and Th2 in PBMCs from EP patients were higher than those in healthy controls, and the Th1/Th2 ratio was dramatically lower than in PV patients (P<0.01). The ratios of IFN-γ/IL-4 and T-bet/GATA-3 in EP patients were both less than 1.0, suggesting a reversal when compared with the other two groups. Our study indicated that the EP patients exerted a Th1/Th2 bidirectional response pattern, and the balance of Th cell subsets inclines to Th2, which might be one of the important mechanisms of EP pathogenesis.

Objective To investigate the incidence of lower urinary tract symptoms(LUTS) and bladder function in the community-dwelling elderly females. Methods The questionnaires were administered,including complaints of LUTS,International Prostate Symptoms Score(IPSS) and quality of life assessment.Meanwhile,uroflowmetry was performed and volume of residual urine was measured. Results The total number of women surveyed was 359.According to IPSS,the prevalence of moderate to severe LUTS(IPSS≥8) was 39.0%.The prevalence in those with age of 50-59,60-69 and ≥70 years were 35.1%,46.2% and 54.8%,respectively(P=0.034).Of all the women surveyed,73.8% had predominantly irritative symptoms.The maximum flow rates(Qmax)were(24.5?11.5) mL/s,(22.7?11.0) mL/s and(14.5?8.2) mL/s,respectively for these age groups(P

Aim To investigate the effect of p-hydroxybenzaldehyde ( HD) on intestinal fibrosis in mice based on mouse intestinal fibrosis model and in vitro EMT model,and to explore the underlying mechanism Methods HE staining, Masson staining, immunohisto-chemistry ,qPCR, Western blot and other experimental methods were used to verify the effect of HD on intestinal fibrosis in mice and the potential mechanism. Results In vivo experiments showed that compared with the normal group, the DSS-induced intestinal fibrosis model group had shortened colon, increased colon his-topathological score, increased collagen volume fraction, and significantly increased collagen I expression. After treatment with 4, 10, and 25 mg • kg

Adult ; Aged ; Aged, 80 and over ; Decompression, Surgical ; Female ; Fractures, Bone ; complications ; surgery ; Humans ; Leg Injuries ; complications ; surgery ; Male ; Middle Aged

OBJECTIVERecent studies have shown that integrin linked kinase (ILK) plays an important role in the pathogenesis and development of some kidney diseases. This study aimed to investigate the relationship between ILK and renal glomerular damage in children with Henoch schonlein purpura nephritis (HSPN).

METHODSOne hundred and eighty eight HSPN children (aged 3 to 17 years) were assigned to five groups according to the classification of the International Study of Kidney Disease in Children (ISKDC): grade < or = IIa (n = 62), grade IIb (n = 42), grade IIIa (n = 29), grade IIIb (n = 40) and grade > or = IV (n = 15). Fifteen children with basement membrane nephropathy served as the control group. ILK expression on glomeruli was ascertained by immunohistochemical staining. The relationships of ILK expression on glomeruli with glomerular histopathologic lesions and urinary protein excretions were examined.

RESULTSThe positive areas of ILK expression on glomeruli in the control, grade < or = IIa, grade IIb, grade IIIa, grade IIIb and grade > or = IV groups were (3.35 + or - 1.01)%, (4.88 + or - 1.13)%, (9.64 + or - 1.36)%, (11.27 + or - 1.68)%, (17.42 + or -3.0)% and (20.62 + or - 2.32%), respectively. There were significant differences in the ILK expression between groups (p<0.01). ILK expression on glomeruli increased with increased urinary protein excretions. There were significant differences in the ILK expression in children with different urinary protein excretions (p<0.01).

CONCLUSIONSILK might be involved in the process of renal glomerular histopathologic damage and the production of proteinuria in children with HSPN.

Child ; Child, Preschool ; Female ; Humans ; Immunohistochemistry ; Kidney Glomerulus ; pathology ; Male ; Nephritis ; enzymology ; pathology ; Protein-Serine-Threonine Kinases ; analysis ; Purpura, Schoenlein-Henoch ; enzymology ; pathology

OBJECTIVETo observe the therapeutic effect and safety of perineal, crissal and progenital pruritus treated with acupuncture according to differentiation.

METHODSSelf-control method was applied in these 32 cases. Changqiang (GV 1), Huiyin (CV 1), Qugu (CV 2), Sanyinjiao (SP 6) and Ashi points etc. were punctured as main points, and adjunct points were added according to differentiation: Taichong (LR 3) and Ququan (LR 8) etc. were added for wind and heat excess of liver meridian, Xuehai (SP 10) and Quchi (LI 11) etc. were added for blood deficiency and wind dryness. Itchiness, skin lesions sign scores and therapeutic effects were observed before and after treatment.

RESULTSThe total scores of itchiness before and after treatment were 6.06 +/- 1.46 and 2.19 +/- 1.71 respectively, and the total scores of skin lesions sign were 4.38 +/- 2.21 and 1.50 +/- 1.44, indicating that the scores and the total scores of itchiness and skin lesions sign reduced obviously after treatment (P < 0.05, P < 0.01); the cured and markedly effective rate was 73.4% (11/15) for wind and heat excess of liver meridian, and 70.6% (12/17) for blood deficiency and wind dryness, presenting similar therapeutic effect (P > 0.05). Hematoma or ecchymosis appeared in 2 cases, and disappeared spontaneously after 2-3 days, without obvious adverse reaction.

CONCLUSIONSimple perineal, crissal and progenital pruritus treated with acupuncture according to differentiation is effective, safe and applicable.

Acupuncture Therapy ; Adolescent ; Adult ; Female ; Humans ; Male ; Middle Aged ; Pruritus Ani ; therapy ; Pruritus Vulvae ; therapy ; Treatment Outcome ; Young Adult

As one of the most serious types of psoriasis, pathogenesis of erythrodermic psoriasis (EP) is unclear so far. In this study, we aimed to detect the levels of Th1/Th2 cytokine-associated transcription factors and T-lymphocyte clone in peripheral blood mononuclear cells (PBMCs) derived from EP patients, and gene expression level of T-bet/GATA-3 in skin lesion. The potential role of Th1/Th2 reaction pattern played in the pathogenesis of EP was also discussed. Serum levels of IFN-γ, IL-2, IL-4 and IL-10 were quantified by ELISA among 16 EP patients, 20 psoriasis vulgaris (PV) patients and 15 healthy controls. The expression levels of T-bet/GATA-3 in the skin lesion and PBMCs were examined by real-time qPCR. The ratio of Th1/Th2 was measured by flow cytometry. The levels of IFN-γ, IL-2, IL-4 and IL-10 were higher in EP patients than in the healthy controls. The levels of IL-4 and IL-10 were 69.44±11.45 and 12.62±4.57 pg/mL, respectively, in EP patients, significantly higher than those in PV patients and healthy controls (P<0.05). Flow cytometry revealed the levels of both Th1 and Th2 in PBMCs from EP patients were higher than those in healthy controls, and the Th1/Th2 ratio was dramatically lower than in PV patients (P<0.01). The ratios of IFN-γ/IL-4 and T-bet/GATA-3 in EP patients were both less than 1.0, suggesting a reversal when compared with the other two groups. Our study indicated that the EP patients exerted a Th1/Th2 bidirectional response pattern, and the balance of Th cell subsets inclines to Th2, which might be one of the important mechanisms of EP pathogenesis.
Adult ; Cytokines ; immunology ; Dermatitis, Exfoliative ; immunology ; pathology ; Female ; Gene Expression Regulation ; immunology ; Humans ; Male ; Psoriasis ; immunology ; pathology ; Skin ; immunology ; pathology ; Th1 Cells ; immunology ; pathology ; Th2 Cells ; immunology ; pathology

OBJECTIVETo observe the protective effect of serum with Tongqiao Huoxue decoction (TQHXD) on PC12 cells damaged by glutamate(Glu) and provide clinical proof of the formulae.

METHODSprague Dawley rats underwent intragastric administration of Tongqiao Huoxue decoction twice a day for five days, the administration dose for rats was 4 g x kg(-1). Thus the serum containing TQHXD was prepared. The model of neurocytes damaged by Glu had been established by adding 12.5 mmol x L(-1) Glu to culture medium of PC12 cells. Photic microscope had been used to observe the morphologic changes of cells, the proliferation and activity of PC12 cells had been detected by MTT. Cell membrane permeability had been investigated by the methods of the coloration of trypan blue and AO/EB, the activities of LDH and the contents of NO in the supernatant of PC12 cells had been detected after 5%, 10%, 20% TQHXD being added to the culture medium of PC12 cells damaged by Glu.

RESULTThe morphologic changes in cells were observed under the inverted microscope. Normal PC12 cells in control group were full and bright, but the cells exposed to Glu were shrank; moreover, some cells were disrupted, configuration of the cells in the TQHXD group was closed to that of normal group. Compared with model group, the ratio of living cells in the group being treated by TQHXD after trypan blue staining had a significant increase. After AO/EB staining, cells in TQHXD groups were less being stained by EB. The absorbance (A) values of TQHXD group increased largely. LDH and NO in cell culture supernatant had decreased, it had shown the marked difference.

CONCLUSIONTQHXD showed an apparent protective effect on neurocytes damaged by Glu on proliferation activity and membrane permeability.

Animals ; Cells, Cultured ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Glutamic Acid ; toxicity ; Male ; Neurons ; drug effects ; Neuroprotective Agents ; administration & dosage ; pharmacology ; Neurotoxins ; toxicity ; PC12 Cells ; Rats ; Rats, Sprague-Dawley ; Serum ; chemistry

Objective: To explore the risk predictive value of lipoprotein-associated phospholipase A2 (Lp-PLA2) on acute coronary syndrome(ACS) and to study the relationship between Lp-PLA2 and the severity of coronary stenosis in ACS patients. Methods:A total of 155 ACS patients admitted in our hospital were enrolled. The patient were divided into 2 groups:AMI (acute myocardial infarction) group, n=49 and UA (unstable angina)group, n=106; in addition, there was a Control group, n=44 subjects with normal coronary angiography (CAG).Blood levels of Lp-PLA2 were examined, CAG was conducted and GRACE score, SYNTAX score,Gensini score were calculated. Based on Grace score, ACS patients were divided into 3 subgroups: Low risk subgroup, Grace score≤108, Mid risk subgroup,Grace score 109-140 and High risk subgroup,Grace score≥140.The above parameters were comparedamong different groups. Results: Compared with UA group and Control group, AMI group had increased blood level of Lp-PLA2, P<0.05. Compared with Low risk subgroup, High risk subgroup had much higher Lp-PLA2, P<0.05. Correlation analysis showed that Lp-PLA2 level was positively related to Gracescore (r=0.301, P<0.001). By SYNTAX score and Gensini score evaluation,Lp-PLA2 levels were similar among different subgroups. Conclusion:Blood level of Lp-PLA2 had certain risk predictive value in ACS patients; while it was not related to the severity of coronary stenosis.