Objective To investigate the effect of penehyclidine (PHCD) on Toll-like receptor 4 (TLR4)mRNA and Toll-like receptor 2 (TLR2) mRNA expression in the lung tissue in rats with acute lung injury induced by lipopolysaccharide (LPS) .Methods Sixty healthy SD rats of both sexes weighing 200-220 g were randomly divided into 5 groups ( n = 12 each) :control group (group C) , LPS group and P1-3 groups. Acute lung injury was induced by intraperitoneal (IP) LPS 8 mg/kg in LPS and P1-3 groups. PHCD 0.3, 1.0 and 3.0 mg/kg were given IP after LPS administration in P1-3 groups. The animals were anesthetized at 6 h after IP LPS. Blood samples were collected for determination of serum TNF-α and IL-6 concentrations ( by ELISA) and then sacrificed, the lungs were immediately removed for determination of TLR4 mRNA and TLR2 mRNA expression (by RT-PCR), and microscopic examination. Results LPS significantly increased TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and IL-6 concentrations. PHCD 1.0 or 3.0 mg/kg significantly inhibited LPS-induced increase in TLR4 mRNA and TLR2 mRNA expression in the lung tissue and serum TNF-α and ILr6 concentrations.The lung histopathologic damage was significantly ameliorated in P2 and P3 groups as compared with group LPS.Conclusion PHCD can protect the lungs against LPS-induced acute lung injury through inhibiting TLR4 mRNA and TLR2 mRNA expression in the lung tissue and reducing the inflammatory response.

Objective To evaluate the relationship between spectinomycin resistance in Neisseria gonorrhoeae and mutations in the rpsE gene.Methods Genomic DNA was extracted from 4 clinical isolates of Neisseria gonorrhoeae with different levels of spectinomycin resistance.Then,PCR was performed to amplify the entire rpsE gene and the spectinomycin resistance-determining region (SRDR) in the 16S rRNA gene followed by direct sequencing.Two spectinomycin-sensitive Neisseria gonorrhoeae strains were transformed with the genomic DNA containing the mutant rpsE gene.Subsequently,the susceptibility of the transformants to spectinomycin was determined,and PCR was performed to amplify the rpsE and 16S rRNA genes in the transformants followed by sequencing.Results All the 4 spectinomycin-resistant Neisseria gonorrhoeae strains harbored an A70C transversion in the rpsE gene,but no abnormality in the SRDR of the 16S rRNA gene.No mutations were detected in the spectinomycin-sensitive Neisseria gonorrhoeae strains.The A70C transversion in the rpsE gene was also detected in the two Neisseria gonorrhoeae transformants with spectinomycin resistance.Conclusion The A70C point mutation within the rpsE gene is associated with spectinomycin resistance in Neisseria gonorrhoeae.

As a part of modern medical practice,medical treatment facility(MTF) has playing an important role in disease diagnosis and medical treatment;therefore,nowadays more attention is paid to the medical equipment management.The methods of equipment management which combine actuality and practical experience are introduced in order to make the management of medical equipment more scientific and effective.

Objective To evaluate the reproducibility of quality intima-media thickness(QIMT) and quality arterial stiffness (QAS) technique in assessment of carotid under different measuring methods.Methods Between December 2012 and January 2014,carotid QIMT and QAS examinations were carried out in 30 health volunteers.QIMT and QAS indicators included IMT in QIMT and distensibility coefficient (DC),compliance coefficient(CC),stiffness index α(α),stiffness index β(β),pulse wave velocity(PWV) in QAS.The measurement employed unilateral/once,bilateral/twice,and unilateral/twice methods.Using intra observer and inter-observer variability,the reproducibility was compared between different QIMT and QAS indicators and measuring methods.Results Extremely high level of intra-observer reproducibility was found for both QIMT and QAS technique (ICC＞0.8).QIMT also showed an excellent inter observer reproducibility (ICC＞0.8).In contrast,the reproducibility of QAS technique varied in different indicators (PWV ＞ β ≈ α ＞ CC ＞ DC) and method ( unilateral/once ＞ bilateral/twice ＞ unilateral/twice).Conclusions QIMT measurement was highly reproducible.Whereas the reproducibility of QAS technique varied in different indicators and methods.Due to low reproducibility,the study result did not support the clinical application of DC indicator and unilateral/once method.

Objective To investigate the effects of triptolide on proliferation,apoptosis and the changes of Ski,Smad3,Smad7 and collagen type I(ColI) in cultured rat mesangial cells induced by transforming growth factor (TGF)-β1.Methods Cultured HBZY-1 rat mesangial cells were divided into 5 groups:(1)normal control group; (2)TGF-β1 group (10 μg/L); (3)-(5)triptolide (0.4,2,10 μg/L)+TGF-β1 (10 μg/L) groups.The cell proliferation was detected by MTT.Apoptosis of mesangial cells was detected by TUNEL assay.The expressions of Ski,Smad3,Smad7 mRNA were examined by real-time quantitative PCR.The expressions of Ski,Smad3,Smad7 and ColI protein were detected by Western blotting.The localizations of Ski and Smad3 protein were detected by laser confocal fluorescence microscope.Results Compared with the normal control,TGF-β1 (10 μg/L) significantly stimulated mesangial cells proliferation,while decreased apoptosis.The mRNA and protein expressions of Ski,Smad7,Smad3 and ColI protein expression in TGF-β1 group were increased (P ＞0.05).In comparison with TGF-β1 group,triptolide could significantly inhibit TGF-β1-induced mesangial cells proliferation in dose-dependent manner,and promote the apoptosis of mesangial cells.In TGF-β1 group,mRNA and protein expresscons of Ski and Smad7 were increased (P＜0.05),Smad3 mRNA and protein were decreased (P ＞0.05),and ColI protein was decreased (P＜0.01).In comparison with TGF-β1 group,fluorescence intensity of Ski,Smad3 proteins was significantly increased in cytoplasm,while decreased in nucleus.Conclusions Triptolide can inhibit TGF-β1-induced mesangial cells proliferation through regulating the expressions of Ski,Smad7 mRNA and protein,inhibiting Ski.Smad7 translocation to the nucleus,and down-regulating Smad3 mRNA and protein expression.Triptolide can promote apoptosis of mesangial cells.

Objective To investigate the expression of triggering receptor expressed on myeloid cells receptor-1 (TREM-1) in plasma and bronchoalveolar lavage fluid (BALF) and its correlation with Galactomannan,IFN,IL-6 and IL-10 in Aspergillus infected mice.Methods Cyclophosphamide (CTX) was intraperitoneally injected and fumigatus spore suspension was inhaled by nose to establish the immunocompromised invasive pulmonary aspergillosis (IPA) mouse model.Healthy controls,immunocompromised only and IPA only groups were also established.Each group had 6 mice.After inoculation,mice were sacrificed.Lung tissue specimens,BALF,and plasma samples were collected.Plasma and BALF soluble TREM-1 (sTREM-1),Galactomannan,IFNγ,IL-6,and IL-10 were detected by ELISA.Results Positive Aspergillus fumigatus was found by tissue culture in the lung.Infiltration of inflammatory cells,blood congestion and interstitial lung tissue injury were observed in histological sections of both IPA and immunocompromised IPA mice.Compared to IPA group [(453.78 ± 74.18) ng/L,P ＜ 0.001;(10.21±1.46) ng/L,P＜0.001] and control group [(245.16 ±65.85) ng/L,P＜0.001;(6.60 ± 3.74) ng/L,P ＜ 0.001],the plasma and BALF sTREM-1 significantly increased in immunocompromised IPA group [(1 537.64 ± 359.52) ng/L;(20.12-± 2.72) ng/L].Compared to control group,both the BALF sTREM-1 in IPA group (P =0.041) and the plasma and BALF Galactomannan,IFNγ,IL-6,and IL-10 levels in IPA and immunocompromised IPA groups were significantly higher (P ＜0.01).Pearson correlation analysis showed that plasma and BALF sTREM-1 were significantly correlated with Galactomannan (r =0.83,P ＜ 0.001;r =0.82,P ＜ 0.001),IFNγ (r =0.79,P＜0.001;r=0.61,P＜0.01),IL-6 (r=0.81,P＜0.001;r=0.66,P＜0.01),andIL-10 (r=0.70,P =0.001;r =0.54,P =0.02).Conclusions Plasma and BALF sTREM-1 appears highly expressed in Aspergillus infected mice.sTREM-1 in mice plasma and BALF is closely correlated with Galactomannan,IFNγ,IL-6,and IL-10 levels,which suggests that sTREM-1 has great diagnostic value during invasive fungal infection.

Objective To investigatethe clinical value of troponin-I(cTnI)in patientswith septic shocka-nd left ventricular diastolic dysfunction. Methods As a retrospective analysis ,38 patients with left ventricular di-astolic dysfunction and septic shock(Sa group),as well as 20 patients with normal cardiac function(Sn group) were enrolled in this study. Moreover ,20 patients with left ventricular diastolic dysfunction and without septic shock were used as control group(Ca group). The ratio of early diastolic mitral inflow velocity to early diastolic mi-tral annulus velocity(E/e′)was measured as the evaluation index of left ventricular diastolicfunction by echocar-diography within 72 hours after admission to ICU. Level of cTnI was detected in all cases and the relationship was evaluated by E/e′. Receiver operating characteristic curve(ROC)was constructed to indicate the predictable value of left ventricular diastolic dysfunction in patients with septic shock. Results The level of cTnI was significantly elevated in both Sa group and Sn group(P<0.05),while the level of cTnI and E/e′in Sa group were significantly higher than those in Sn group(P < 0.05). cTnI was positively correlated with E/e′(r = 0.367 ,P = 0.004). The area under the curve(AUC)of cTnI was 0.834,with the cut-off value of 0.49 ng/mL(sensibility=77.6,specificity=80.7). Conclusion The level of cTnI was significantly higher in patients with septic shock. cTnI was significantly correlated to left ventricular diastolic dysfunction in patients with septic shock. cTnI ≥ 0.49 ng/mL could be an available predictor for left ventricular diastolic dysfunction in patients with septic shock.

Objective To study the clinical and pathologic changes in gigantomast.Methods Tissue sections were prepared from 180 cases of breast hypertrophy and 45 cases of normal breast tissues.The morphological changes and the expression and localization of ERRγwere evaluated on the HE and immunohistochemistry stained sections between hypertrophy and normal breast tissues.Results Compared with normal breast,hypertrophic breast showed expended ducts and obvious hyperplasia of the duct epithelial papillary.Hypertrophic breast tissues demonstrated strong expression of ERR-γ in ducts and lobules.Conclusions Upregulated expression of ERRγ is identified in hypertrophic breast tissues that may associate with the development of gigantomastia.

Oxidation method with sodium iodide was used to synthesize immunogenic antigen (PF-BSA) and coating antigen (PF-OVA) of paeoniflorin. UV spectroscopy showed that paeoniflorin was successfully conjugated with BSA and OVA. After immunized by PF-BSA, the mice can produce anti-paeoniflorin antibodies specifically. The ELISA test results showed the high titer (1:12 800) and specificity (IC50 = 0.791 mg x L(-1)) of the antiserum from mice injected with PF-BSA. Also, the antiserum showed low cross activities against nine traditional Chinese medicine (TCM) of small molecules. These artificial antigens were successfully synthesized and the anti-paeoniflorin antibody well prepared, which provides the experimental basis for the further study of ELISA and its kit.
Animals ; Antibodies ; analysis ; Antigens ; chemistry ; immunology ; Drugs, Chinese Herbal ; chemistry ; Enzyme-Linked Immunosorbent Assay ; Glucosides ; chemistry ; immunology ; Male ; Mice ; Mice, Inbred BALB C ; Monoterpenes ; chemistry ; immunology ; Serum Albumin, Bovine ; chemistry ; immunology

BACKGROUND:Dendritic cel s can regulate the immunological reaction in the intestinal tract, this functional deficit may induce inflammatory bowel disease. Tol-like receptor-4/nuclear factor-κB pathway is highly involved in this reaction. OBJECTIVE:To establish experimental colitis model in rats, to observe effects of resina draconis on dendritic cel s and Tol-like receptor-4/nuclear factor-κB expression in rats with experimental colitis, and to explore its action mechanism. METHODS:A total of 44 male Sprague-Dawley rats were randomly assigned to four groups (n=11):blank control group, model group, resina draconis group, 5-aminosalicylic acid treatment group. With the exception of blank control group, 2,4,6-trinitrobenzenesulfonic acid-induced ulcerative colitis models were established in the model group, resina draconis group and 5-aminosalicylic acid treatment group. After the models were successful y established, the rats in the resina draconis and 5-aminosalicylic acid treatment groups were intragastrical y treated with resina draconis [(0.75 g(kg·d)] and 5-aminosalicylic acid [100 mg(kg·d)] respectively for 10 days. RESULTS AND CONCLUSION:Disease activity index, macroscopic colonic damage score and histopathological score were significantly decreased in the resina draconis group compared with the model group (P<0.05). Symptoms and tissue damages were obviously lessened in the 5-aminosalicylic acid treatment and resina draconis groups compared with the model group. Expression rates of CD80 and CD86, as wel as expression levels of Tol-like receptor-4 and nuclear factor-κB were significantly higher in the model group compared with the blank control group, resina draconis group and 5-aminosalicylic acid treatment group (P<0.05, P<0.01). Tol-like receptor-4 and nuclear factor-κB expression was significantly lower in the resina draconis group than that in the 5-aminosalicylic acid treatment group. Experimental findings indicate that, resina draconis can partial y relieve experimental colitis symptoms in rats and effectively inhibit the activation of dendritic cel s in the mesenteric lymph node. Resina draconis can relieve enteric inflammatory reaction by suppressing the expression of Tol-like receptor-4 and nuclear factor-κB in rats.