Animals ; Drugs, Chinese Herbal ; chemistry ; Humans ; Medicine, Chinese Traditional ; Microscopy, Confocal ; Muscle, Smooth, Vascular ; ultrastructure

Objective: To observe the effect of electroacupuncture (EA) on Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) pathway in knee joint synovial tissues of rats with rheumatoid arthritis (RA) and to explore the action mechanism of EA on RA. Methods: Twelve of the 48 SPF male Sprague-Dawley (SD) rats were assigned to a normal group by the random number table method. The remaining 36 rats were subjected to RA model preparation by intradermal injection of the Freund's complete adjuvant into the right hind foot pad of each rat under sterile conditions. After the model was successfully prepared, rats were then divided into a model group, a drug group and an EA group according to a random number table method (n=12). Rats in the drug group were treated with 2 mL aqueous solution of tripterygium glycosides [8.1 mg/(kg?bw)]; rats in the EA group were treated with EA at bilateral Yanglingquan (GB 34) and Zusanli (ST 36), for 30 min each time; rats in the normal group and the model group were placed in a special rat fixation tank for 30 min each time, and received the same dose of normal saline as those in the drug group. Rats in all groups received intervention once a day for 4 weeks. Diameter of rat ankle joint and rat arthritis index were measured before and after the intervention. At the end of the experiment, the expressions of phospho-JAK2 and phospho-STAT3 were determined by immunohistochemistry. Quantitative real-time polymerase chain reaction (RT-qPCR) was used to detect JAK2 and STAT3 mRNAs expressions. Results: After the model was produced, the arthritis index >2 was considered successful in model preparation. Compared with the model group, the ankle joint diameters and arthritis indexes of rats in the drug group and the EA group were significantly lower (all P<0.01); immunohistochemical staining cells with phospho-JAK2 and phospho-STAT3 were significantly decreased (all P<0.01); the expression levels of JAK2 and STAT3 mRNAs were decreased with statistical differences (all P<0.01). There were no significant differences between the EA group and the drug group (all P>0.05). Conclusion: EA can alleviate the inflammatory response of RA rats, improve their pathological conditions, reduce the expressions of phospho-JAK2 and phospho-STAT3 in the synovial tissue of knee joint, and decrease the expressions of JAK2 and STAT3 mRNAs. The therapeutic effect of EA is comparable to that of the tripterygium glycosides. The mechanism of EA treatment may be related to the inactivation of the JAK2/STAT3 pathway.

Objective To investigate the diagnosis value of the cytological examination of the cerebrospinal fluid of central nervous system leukemia.Methods Adopting cell smear centrifugal machine to collect the cerebrospinal fluid cells,the cells were stained and examinated under the microscope.Results Fifty-nine children with different type of leukemia had been examinated by 438 times by cerebrospinal fluid.The positive rates of the cases and samples were 15.3% and 8.7%,respectively.Conclusion The cytological examination of cerebrospinal fluid is especially valuable for the early diagnosis ,therapy and relapse of central nervous system leukemia of monitoring.

Background Vascular endothelial growth factor(VEGF) has been shown to be associated with the pathogenesis of age-related macular degeneration (AMD),therefore VEGF is a target for the treatment of wet AMD.However,the mechanism of VEGF in the pathogenesis of AMD is not clearly understood.Studying the correlation between VEGF gene polymorphism and AMD is becoming a new research hotspot,but relevant studies on Han Chinese have not been performed.Objective This study was to investigate the association between the VEGF +936C/T gene polymorphism and AMD in the Chinese population.Methods A pilot prospective and nonrandomized controlled trial was designed.This protocol complied with Declaration of Helsinki and was approved by the Ethic Committee of Chinese PLA Second Artillery General Hospital.Written informed consent was obtained from each subject prior to entering the study.Two hundred AMD patients and 200 age-and gender-matched normal controls were enrolled in this study.The genomic DNA was extracted from the peripheral blood samples of the subjects,and analysis of the VEGF polymorphisms at the +936 position in the promoter and 3＇-untranslated regions was performed by the restriction fragment length polymorphism method.Frequencies of the VEGF+936C/T genotype were compared between the two groups,and the risk of the VEGF+936C/T gene polymorphism in pre-disposing AMD was evaluated.Results No significant differences were seen in the incidence rates of smoking(P ＝ 0.76),hypertension(P ＝ 0.84),hyperlipidemia (P=0.71),diabetes mellitus (P=0.86) and cardiovascular disease(P=0.89) between the AMD group and the normal control group,and BMI was matched between the two groups (P =0.18).The prevalence of the TT genotype was 9.0％ (18/200)in the AMD group,but that in the normal control was 3.5％ (7/200),showing a significant difference between the two groups (P =0.03).The odds ratio (OR) was 2.73 with a 95％ confidence interval(CI) of 1.11 to 6.68 for AMD in this genotype.The CC and CT genotypes were not significantly different between the two groups (P =0.52,P =0.57).The genotype frequency and allele frequency conformed to HardyWeinberg equilibrium law.There were no significant differences found in the CC,CT,TT genotype frequencies among the early AMD,geographic atrophy AMD and choroidal neovascular AMD (all at P＞0.05).Conclusions The VEGF+936TT genotype is associated with AMD in Han Chinese population.

Objective To discuss causes and corresponding prevention for groin pain occurred early after total hip arthroplasty(THA).Methods A retrospective study was done on 189 cases(193 hips)treated with THA including unilateral procedures in 185 hips and bilateral procedures in eight hips to analyze common causes for early groin pain.Results Groin pain was found in 9.3% hips(18/ 193)during hospital stay,including 1.6%(3 cases)with deep infection,1.6%(3 cases)with incision infection,1%(2 cases)with posterior dislocation,4.1%(8 cases)with leg lengthening and 1%(2 cases)with hematoma.Conclusions Despite of the numerous diagnostic alternatives available to the orthopedic surgeon,detailed history,careful physical examination,necessary laboratory and imaging stud- ies can contribute to a correct determination of causes for groin pain.Meanwhile,appropriate indication, accurate preoperative radiographic measurement,intraoperative standardized surgical procedures and per- fect rehabilitation are necessary to avoid complications.

OBJECTIVE:To establish a RP-HPLC method for simultaneous determination of chlorapheniramine maleate,furacilin and ephedrine hydrochloride in Puma nose drops.METHODS:The analysis was carried on a XDB C 8 column;the mo?bile phase was composed of methanol(A),acetonitrile(B)and0.02mol/L potassium dihydrogen phosphate solutions(containing0.2%triethylamine and adjusted to pH3.0with phosphoric acid,C)with linear gradient elution(0min～3.5min,A∶B∶C=6∶13∶18,8.5min,A∶B∶C=6∶30∶64)and the flow rate was1.0ml/min;the detection wavelength was254nm and the column temperature was30℃.Chloramphenicol was used as the internal standard.RESULTS:The linear ranges were0.04～0.20mg/ml for chlorapheniramine maleate,0.02～0.10mg/ml for furacilin,0.50～2.50mg/ml for ephedrine hydrochlo?ride.The average recoveries were99.44%(RSD=0.48%,n=3)for chlorapheniramine maleate,101.36%(RSD=0.41%,n=3)for furacilin and99.43%(RSD=0.59%,n=3)for ephedrine hydrochloride.CONCLUSION:The method is reliable,accurate and suitable for quality control of Puma nose drops.

Objective: To study the chemical constituents in the whole herb of Balanophora involucrate. Methods: The compounds were isolated and purified using polyamide, silica gel colimu, ODS, MCI gel, and semi-preparative HPLC, and their structures were elucidated by means of physicochemical properties and spectroscopic analysis. The anti-inflammatory activities of all the isolated compounds were evaluated using Griess method and ELISA for the determination of LPS-induced NO and IL-6 releases in inflammation cell model induced by LPS. Results: Eleven lignans were isolated from 75% ethyl alcohol extract from the whole herb of B. involucrata and identified as (+)-pinoresinol (1), (+)-5’-hydroxypinoresinol (2), isolariciresinol 4-O-β-D-glucopyranoside (3), (+)-isolariciresinol (4), burselignan (5), (+)-9-acetoxyisolariciresinol (6), yunnanensin A (7), (-)-secoisolariciresinol-4-O-β-D- glucopyranoside (8), (-)-secoisolariciresinol (9), dihydrocubebin (10), and secoisolariciresinol-9’-acetate (11). Conclusion: Among them, compound 11 is a new natural product, and compound 2, 5, 7, 8, and 10 are isolated from the genus of Balanophora for the first time. All compounds showed strong anti-inflammatory activities.

Objective To investigate whether leukotriene D4 (LTD4) regulates eotaxin-3 (Eot-3) expression in bronchial epithelial cells, and study effect of pranlukst on the regulation.Methods BEAS-2B cells and normal human bronchial epithelia cells were pre- treated with LTD4 for 1 hour,stimulated with interleukin-4, the cells were incubated for 24 hours. Eot-3 protein in supernatant were measured by enzyme linked immunosorbent assay(ELISA). The cells were pretreated with pranlukast in different concentration, then the above procedure was repeated. Results The untreated bronchial epithelial cell expressed Eot-3 protein on a very low level. After stimulating with IL-4 and incubating for 24 hours, Eot-3 production increased significantly. Pretreating the cells with LTD4 enhanced the inducing effect of IL-4. Pranlukast inverted the upregulation of LTD4. Conclusions Upregulating the expression of Eot-3 induced by IL-4 on bronchial epithelial cells may explain partially the mechanism of leukotrienes involving airway allergic inflammation of asthma. The invertion impact on upregulation of LTD4 by pranlukast may be one of mechanisms that leukotrienes receptor antagonist cure asthma.

Objective To investigate the effects of different concentrations of sodium fluoride on the morphologic characteristics of primarily cultured thyroid cells of SD rats and in order to obtain important proof for approaehing the mechani8m of thyroid gland damage caused by fluoride.Methods Thyroid cells of SD rat were primarily culture for 96 hours,and cell density was adjusted to 5.0×108/L Cell suspension with 5 ml Wills seeded into 6 weII plates,after 12 hours,0(contr01),10.100,1000 μmol/L of sodium fluoride was added into the well, witll each well representing different level of treatment group.Finally the cultured thyroid cells were collected for morph010gic study.Results Under microscope,the transparency of the control thyroid cells Was good,and cells gathered in cluster and adhered to wall.But a lot of cells treated with fluoride suspended,and lost their transparency-under scaning delectron microscope,the control calls showed integrated membrane and tightness to each other,as well as clear boundary between cells normal proliferation.While the thyroid cells treated with 10,100 μmol/L sodium fluoride 0bviouslv shrinked and deformed,and the cells treated with 1000 μmol/L of sodium fluoride were broken-Conclusions nuoride can affect the growth and development of thyroid cell and damage the structure and morphology.Sodium fluoride affects the morphologie characteristics of thyroid cells in a dose-response manner.

Objective To explore the effects of thiamine and riboflavin on H2O2-induced DNA oxidative damage in human umbilical vein endothelial cell line ECV304.Methods ECV304 cells were incubated with 10,100,500,1000 mg/L of thiamine or 20,100,300,500 nmol/L of riboflavin for 24 h,and then oxidative damage of cells were induced by 25 mol/L H2O2 for 30 min.DNA damage was detected with single cell gel electrophoresis(SCGE)assay.ECV304 cells incubated without H2O2,thiamine and riboflavin were served as negative controls,and those incubated with H2O2 and without thiamine and riboflavin were served as positive controls.Results H2O2 induced DNA damage,and the indices of percent of DNA damage cells,percent of tail DNA,tail length and Olive tail moment were increased.The indices of cells pretreated with 10,100,500 mg/L of thiamine or 20,100,300 nmol/L riboflavin were significantly decreased(P0.05).Conclusion Proper supplementation of thiamine and riboflavin may decrease H2O2-induced DNA oxidative damage,while excess thiamine and riboflavin supplementation may be harmful to DNA and enhance the susceptibility to H2O2 potentially.