[ ABSTRACT] AIM:To investigate the biological characteristics of newborn rabbit tracheal chondrocytes in vitro. METHODS:Newborn rabbit tracheal chondrocytes were obtained by the method of enzyme digestion, and then cultured in monolayer in vitro.Morphological and growth observations were performed under inverted phase contrast microscope.The ultrastructures of the cells were observed under scanning electron microscope and transmission electron microscope.The bi-ological characteristics of secreted extracellular matrix components were detected by real-time PCR, immunocytochemistry staining and toluidine blue staining.RESULTS: Newborn rabbit tracheal chondrocytes isolated and cultured in vitro showed short triangular or irregular shapes, and adherent growth very well.The ultrastructures of the cells showed pore and abundant cytoplasm and organelles, with a lot of protein secretions in the cells.The chondrocytes expressed the mRNA of collagen I, collagen II and proteoglycans, mainly collagen II and proteoglycans.Immunocytochemistry staining showed col-lagen II and SOX9 positive, and collagen I weakly positive.Toluidine blue staining was also positive.CONCLUSION:Enzyme digestion and monolayer culture are suitable method to obtain newborn rabbit tracheal chondrocytes.These cells, secreting extracellular matrix components, are able to be selected as seed cells for tissue engineering of trachea in vitro, and used to study the therapeutic method for neonatal rabbit tracheal stenosis.

OBJECTIVE:To analyze the features of risperidone-induced leucocytopenia ADR.METHODS:Twenty-one ADR cases of risperidone-induced leucocytopenia reported by our hospital were collected and analyzed during 2004-2015.The characteris tics of risperidone-induced leucocytopenia were discussed.RESULTS:Among 21 patients,there were 10 male and 11 female.The age was from 15 to 72 years old.Nine cases of patients were 31-40 years old (42.9％).Most of the original disease was schizophre nia.Incubation period of leucocytopenia caused by risperidone was (28.6 ± 21.4) d.Patients had no discomfort complain when leucocytopenia occurred.The lowest white blood cells reported was(3.1 ± 0.5)× 109 L-1.The leucocytopenia were improved after reduc tion,drug withdrawal and symptomatic treatment.CONCLUSIONS:Patients usually have no body discomfort complain when risperidone-induced leucocytopenia appears.Doctors should moniter ADR regularly,identify it earlier and treat carefully.

Aim The effects of midazolam on the whole-cell sodium currents in rat sympathetic neurons were studied to explore the mechanisms where by midazolam mediates hypotension. Methods Whole-cell patch-clamp recordings were performed on enzymatically isolated rat superior cervical sympathetic neurons. Results Midazolam dose-dependently blocked the whole-cell sodium currents evoked by a voltage step to 0 mV from a holding potential of -80 mV with a mean drug concentration required to produce 50% current inhibition (IC50) values of 18.35 ?mol?L-1; Clinically relevant concentration of midazolam(0.3 ?mol?L-1) reduced sodium peak currents by 19.98%(P

BACKGROUND: Bone marrow mesenchymal stem cells are important seeded cells for construction of tissue-engineered trachea, but there is no special surface marker. Therefore, identification of bone marrow mesenchymal stem cells is mostly based on morphology, phenotype antigen and the function of differentiation. OBJECTIVE: To explore the feasibility of the tracheal chondrogenic differentiation of bone marrow mesenchymal stem cells under a special condition through isolation, cultivation and identification of bone marrow mesenchymal stem cells. METHODS: Rabbit bone marrow was acquired in the sterile environment to isolate and culture bone marrow mesenchymal stem cells to passage 2 by bone marrow adherence and screening method. Flow cytometry identified the phenotype CD44, CD45 of bone marrow mesenchymal stem cells at passages 1 and 2. Rabbit tracheal samples were acquired in the sterile environment, the tracheal chondrocytes were isolated and cultured by enzyme digestion, and toluidine blue staining was used to detect aggrecan. Bone marrow mesenchymal stem cells were co-cultured with tracheal chondrocytes by Transwel and transforming growth factor β1. Cel morphology was detected under an inverted microscope. Real-time quantitative PCR and toluidine blue staining detected the extracel ular matrix components, such as type Ⅱ col agen and aggrecan.RESULTS AND CONCLUSION: After isolation and culture, cells were spindle and irregular in morphology, and passaged cells thrived that were gathered into a fish-like colony growth. For passage 1 bone marrow mesenchymal stem cells, the positive rates of phenotype antigen CD44 and CD45 were respectively 96.97% and 13.72%; for passage 2 cells, the positive rates of phenotype antigen CD44 and CD45 were 99.11% and 8.54%, respectively. Tracheal chondrocytes were positive for toluidine blue staining. The morphology of induced bone marrow mesenchymal stem cells changed from long fusiform to triangular or irregular shape, indicating the chondrocytes expressed type Ⅱ col agen and aggrecan, and toluidine blue staining was positive. These results showed bone marrow adherence and screening method could acquire bone marrow mesenchymal stem cells, and the purity of passage 2 cells is higher. Under a special condition, bone marrow mesenchymal stem cells have the potential of chondrogenic differentiation, and can be selected as seed cells for construction of tissue-engineered trachea.

Objective To determine whether Morphine has the ability to enhance HIV-1 replication in MT2 and Macrophage in vitro and assess the influence of Naloxone on Morphine2s effect.Methods MT2 cells were randomly assigned into 4 groups: (1) Morphine treatment for MT2 group, (2) Morphine+Naloxone co-treatment for MT2 group, (3) Naloxone treatment for MT2 group and (4) MT2 Control;Macrophages were also randomly assigned into 4 groups: (5) Morphine treatment for Macrophage group, (6) Morphine+Naloxone co-treatment for Macrophage group, (7) Naloxone treatment for Macrophage group and (8) Macrophage Control. Group (2), (3), (6) and (7) were pre-treated with 10-8 mol/L Naloxone for 0.5 h, and then group (1) and (2) were treated with 10-12, 10-10 and 10-8 mol/L Morphine for 24 h;group (5) and (6) were disposed of 10-10 mol/L Morphine for 24 h.All 8 groups were added in HIV-1 viral strain with 50% tissue culture infective dose(TCID50).P24 antigen in MT2 cells culture supernatant at day 3, 4, 5 and 6, and in Macrophages culture supernatant at day 4, 6, 8, 10 and 12 after infection were determined with ELISA.Student2s t-test and ANOVA were used to compare the differential expression in different groups, and repeated measures ANOVA was used to compare the increasing or decreasing expression of p24 antigen in morphine treatment groups than that in the control group at different time points.Results On the 3rd day of infection with HIV-1 in MT2 cells, the expression of p24 antigen in 10-12, 10-10 and 10-8mol/L dose of group (1) were (4.44?.30), (5.59?.25) and (4.60?.24) ng/ml respectively, compared to control[(1.93?.05) ng/ml, t= 14.15, 24.74 and 19.14, all P<0.01].On the 4th day, 10-12, 10-10 and 10-8mol/L dose of group (1) resulted in a significant increase of p24 antigen expression [(24.30?.66), (31.73?.17) and (26.02?.37) ng/ml]in culture supernatants compared to control[(8.03?.09) ng/ml, t=10.59, 34.92 and 81.2, all P<0.01].On the 5th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (1) were (56.30?.26), (81.77?.49) and (63.66?.57) ng/ml respectively, compared to control [(15.30?.91) ng/ml, t= 45.83, 43.51 and 30.07, all P<0.01].On the 6th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (1) were (150.70?.97), (243.09?.93) and (173.72?.73) ng/ml respectively, compared to control [(41.01?.84) ng/ml, t= 21.09, 39.02 and 29.55, all P<0.01].The enhanced multiple of p24 antigen expression in three doses of morphine treatment group compared to control increased with HIV-1 infected MT2 cells time, trend analysis of repeated measurements showed statistically significant time effect (F=842.18, P<0.01). On the 4th day of infection with HIV-1 in Macrophage cells, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (5) were (0.68?.15), (0.87?.41) and (0.75?.09) ng/ml respectively, compared to control [(0.60?.01) ng/ml, t= 7.27, 11.06 and 3.02, all P<0.05]. On the 6th day, 10-12, 10-10 and 10-8 mol/L dose of group (5) resulted in a significant increase of p24 antigen expression[(1.64?.57) , (2.07?.12 ) and (1.75?.17) ng/ml]in culture supernatants compared to control [(1.16?.07) ng/ml, t=8.93, 11.3 and 5.45, all P<0.01].On the 8th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of group (5) were (6.31?.17), (8.81?.34) and (7.19?.11) ng/ml respectively, compared to control [(3.84?.45) ng/ml, t=8.83, 15.11 and 12.42, all P<0.01]. On the 10th day, the expression of p24 antigen in 10-12, 10-10 and 10-8 mol/L dose of Morphine treated group were (32.30?7.55), (50.74?7.55) and (39.74?.56) ng/ml respectively, compared to control [(17.55?.86) ng/ml, t= 13.65, 17.84 and 36.69, all P<0.01].The enhanced multiple of p24 antigen expression in three doses of group (5) compared to control increased with HIV-1 infected Macrophage cells time, trend analysis of repeated measurements showed statistically significant time effect (F=135.58, P<0.01).Conclusions Morphine has the ability to enhance HIV-1 replication in MT2 cell and Macrophage. This Morphine-mediated increase of p24 antigen expression can be blocked by Naloxone.

Qigui Tongfeng tablet (QLTFT) is a traditional Chinese medicine with good effect for treating gout. Here, network pharmacology method and molecular similarity analysis were utilized to study the effective substance basis and molecular mechanism of the QLTFT on the gout. The similarity to the medicinal compounds is reflected in the Tanimoto coefficient that gives the structural similarity of two compounds. Operationally, similar modifiers were described as pairs of concepts with a similarity score of 0. 500. The results of the molecular similarity analysis suggested that the flavonoids in QLTFT could be new leads for gout. Furthermore, complex biological systems may be represented and analyzed as computable networks. Two important properties of a network were degree and betweenness. Nodes with high degree or high betweenness may play important roles in the overall composition of a network. And the results of network analysis showed that dongbeinine, verticinone-N-oxide, verticine N-oxide, peimine, peiminine, isobaimonidine, dongbeirine, peimisine and simi-arenol which with high degree acted on xanthine dehydrogenase/oxidase, matrix metalloproteinase-9, an arachidonate 5-lipoxygenase-activating protein, tyrosine-protein kinase and etc. Inhibition of these targets can prevent the formation of uric acid, reduce inflammation by uric acid and regulate the body's immune response. Thus, these compounds may be the main effective substance basis. The research results not only reveals its molecular mechanism, but also provide a theoretical basis for the quality control of drugs and clinical application.
Gout ; drug therapy ; Humans ; Medicine, Chinese Traditional ; Pharmacology ; methods ; Tablets ; Technology, Pharmaceutical ; methods

Objective To investigate the correlation between the indications,findings,interventions of fibrobronchoscopy(FB) in neonates and their correlative diseases with neonatal FB results and clinical data.Methods Retrospective case series of 243 consecutive patients of 28 days old or younger were investigated underwent FB for the first time from January 2010 to December 2014,at a tertiary care hospital.The common indications for FB and detection rate of respiratory tract diseases were collected.If the findings of FB had significant associations with premature birth and other diseases were analyzed.Associations between interventions and basic illnesses were also analyzed.Results Of the 243 patients undergoing 275 procedures of FB,201 cases were boys(73.1％).The age of FB was (13.34 ± 9.76) days and the weight was (3.08 ± 0.68) kg.Forty-five cases were premature infants (16.4％).A total of 254 procedures were found to have congenital diseases (92.4％),and 177 cases of them had congenital heart diseases (CHD) (64.4％).Common indications for FB were dyspnea(140 cases,50.9％),tachypnea(82 cases,29.8％),and stridor(71 cases,25.8％).A total of 188 upper airway lesions were found and the most common findings were laryngomalacia(56 cases,20.4％) and vocal cord paralysis(bilateral/unilateral,50 cases,18.2％).A total of 315 lower airway lesions were found and the most common findings were airway mucosal inflammation (98 cases,35.6％),trachea and main bronchial stenosis (73 cases,26.5 ％).A total of 21 cases (7.6％) underwent supraglottoplasty during or after FB,while 17 cases (6.2％) underwent tracheal dilation and 10 cases (3.6％) underwent tracheotomy.Compared with non-CHD neonates,neonates with CHD were statistically significantly less likely to have congenital lesions statistically,such as laryngomalacia(15.8％ vs.28.6％,P =0.012),bilateral vocal cord lesions(6.2％ vs.21.4％,P =0.000) and congenital laryngeal dysplasia(0 vs.7.1％,P =0.001).The tracheotomy(0 vs.10.2％,P =0.000) and supraglottoplasty(2.3％ vs.17.3％,P =0.000) were more rare.Nevertheless,they were more likely to have secondary lesions such as the left main bronchial stenosis caused by extrinsic compression (23.7％ vs.1.0％,P =0.000),abnormal bronchial anatomy(9.6％ vs.2.0％,P =0.018),left vocal cord paralysis(9.0％ vs.1.0％,P =0.008) and airway mucosal inflammation(41.8％ vs.24.5％,P =0.004).The tracheostenosis and main broncial stenosis (37.3％ vs.7.1％,P =0.000) with long-term intubation(78.5％ vs.58.2％,P =0.000) were more common.There was no significant difference between term neonates and premature infants in the detection rate of respiratory tract diseases (P ＞ 0.05),tracheotomy (0 vs.4.3 ％,P =0.322),supraglottoplasty (13.3 ％ vs.6.5 ％,P =0.205) or long-term i ntubation (80.0％ vs.69.6％,P =0.157).Complications caused by procedure were rare and mild.Conclusions FB can detect whether the neonates with dyspnea,tachypnea and stridor have laryngomalacia,vocal cord paralysis,airway mucous edema,tracheal and main bronchial stenosis and other signs,and FB may play an important role in diagnosis,treatment and prognosis evaluation of neonatal respiratory diseases.

The aim of this study was to investigate the anti-inflammatory effect of Guizhi Fuling capsule and its active complex (consistent of 15 active compounds) on LPS-induced RAW264. 7 cells. The effect of Guizhi Fuling capsule and its active complex on cell viability in RAW264. 7 cells were determined by MTT assay. The inhibitory effect of Guizhi Fuling capsule and active complex on the releasing of IL-1β, TNF-α and PGE2 induced by LPS in RAW264. 7 cells was detected by ELISA assay. The expression of IL-1β and mPGES-1 in Guizhi Fuling capsule or active complex treated RAW264. 7 cells was examined by Western blot assay. Guizhi Fuling capsule and active complex showed no significant effect on the cell viability in RAW264. 7 cells at doses range from 12.5 to 400 mg x L(-1). Compared with LPS treated group, Guizhi Fuling capsule and active complex dose dependently reduced the releasing of IL-1β, TNF-α and PGE2 induced by LPS in RAW264. 7 cells. Moreover, the expression of IL-1β and mPGES-1 was decreased after Guizhi Fuling capsule and active complex treatment, which might contribute to the inhibitory effect of Guizhi Fuling capsule in the releasing of IL-1β, TNF-α and PGE2. This study provided the evidence that Guizhi Fuling capsule and active complex remarkably inhibited the releasing of IL-1β, TNF-α and PGE2induced by LPS in RAW264. 7 cells by reducing the expression IL-1β and mPGES-1. This study provided an experimental basis of Guizhi Fuling capsule for the treatment of inflammation and a theoretical basis for the development of effective compounds of Guizhi Fuling capsule.
Animals ; Anti-Inflammatory Agents ; pharmacology ; Cell Line ; Cell Survival ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Inflammation ; immunology ; Interleukin-1beta ; immunology ; Macrophages ; drug effects ; immunology ; Mice ; Tumor Necrosis Factor-alpha ; immunology

Atrioventricular Block ; etiology ; Cardiac Catheterization ; adverse effects ; Heart Septal Defects, Ventricular ; therapy ; Hematologic Diseases ; etiology ; Hemolysis ; Humans

OBJECTIVETo explore the effect of new model for tuberculosis (TB) control and management, and provide a scientific basis and justification for making TB control strategies in rural communities.

METHODSAmong those townships with low TB service accessibility by the county TB control institute in Guangxi Xingye county (population of 679 thousands), four townships with total population of 152 518 and inconvenient transportation, were selected as the experimental group to conduct a new model research project.Based on the accessibility for community services, setting diagnosis and treatment management centers in township hospitals, employing family treatment supporters to supervise the treatment process. The TB cases of the base-line and the project expiration of the experimental group were 44 and 117. Meanwhile, three townships including Dapingshan, Longan and Gaofeng in the county with the similar condition and total population of 133 303 were selected as the control group. The control group conducted the provisions of national TB control program in the county TB clinic management. The TB cases of the base-line and the project expiration of the control group were 56 and 110. By double-direction comparison method, the effect of the new model was evaluated through TB patients detection, treatment outcomes and TB control management data. SPSS 13.0 statistical software was adopted and Chi-square test was used for analyzing technical data.

RESULTSAfter two-year project research implementation, in the experimental group the detection rate of new smear-positive TB patients increased from 16.39/100 000 (25/152 518) to 51.14/100 000 (78/152 518) (χ(2) = 27.281, P < 0.01), the cure rate of new smear-positive cases increased from 71.4% (15/21)to 91.1% (51/56) (χ(2) = 4.812, P < 0.05), and the completing treatment rate in newly diagnosed smear-negative cases improved from 23.5% (4/17)to 71.4% (15/21) (χ(2) = 8.622, P < 0.01); the loss rate of newly diagnosed smear-positive cases dropped from 23.8% (5/21) to 0.0% (0/56) (χ(2) = 10.608, P < 0.01), and the loss rate of newly diagnosed smear-negative cases decreased from 64.7% (11/17) to 4.8% (1/21) (χ(2) = 15.624, P < 0.01). Meanwhile, the cure rate of new smear-positive cases in the experimental group, 91.1% (51/56), was higher than the control group, 72.0% (36/50) (χ(2) = 6.531, P < 0.05). The loss rate of newly diagnosed smear-positive cases in the experimental group (0.0% (0/56)) was lower than the control group (16.0% (8/50)) (χ(2) = 7.534, P < 0.01). During the project implementation, in the experimental group the on time rate of taking medicine, 91.5% (107/117) and receiving medicine, 100.0% (117/117), the reexamining sputum ratio, 83.6% (98/117) were higher than that in the control group: 81.8% (90/110), 92.7% (102/110) and 64.5% (71/110). The differences were statistically significant (χ(2) = 4.589, 8.820 and 11.005, P < 0.05).

CONCLUSIONThe new management model had been proved effective. It can improve TB case detection and cure rates, reduce the loss rate of patients, and improve patient treatment and management conditions as well.