OBJECTIVE:To establish the HPLC method for the simultaneous content determination of chloramphenicol and prednisolone acetum in hadro-chlorine gutta.METHODS:The chromatographic column was HiQ sil C 18 ,the methanol-water-glacial acetic acid(70∶30∶0.04)served as mobile phase,and the detecting wavelength was278nm,the flow rate was0.8ml/min and the column temperature was30℃,the sample size was10?l.RESULTS:A satisfactory linear relationship be?tween was found when the concentration determination ranges of chloramphenicol and prednisolone acetum at22.36～89.44?g/ml and25.00～100.00?g/ml respectively;The average recoveries for chloramphenicol and prednisolone acetum were99.65%(RSD=0.88%)and99.57%(RSD=1.02)respectively.CONCLUSION:The method was simple,accurate and good in repro-ducibility,which can be used for the simultaneous contents determination of 2 components in hadro-chlorine gutta.

Objective To observe the effects of dexmedetomidine (DEX) on the expressions of TNF-α, IL-1β and apoptosis-related proteins in rat mesenteric artery.Methods Male SD rats of SPF grade were sacrificed and the mesenteric artery was separated under the stereo-microscope.We established an experimental model of vascular injury induced by lipopolysaccharide (LPS) and randomly divided the injured vessels into dexmedetomidine treatment group and control group.DEX treatment group was divided into 10-8, 10-7, and 10-6 mol/L subgroups according to the different concentrations of DEX.RNA and total protein was extracted in each group.The mRNA expressions of TNF-α, IL-1β and CaSR were detected by RT-PCR and the protein expression of TNF-α, Caspase-3 and AMPK were tested by Western blot.Results DEX (10-8, 10-7, and 10-6mol/L) obviously reduced vascular inflammatory reaction induced by lipopolysaccharide, reduced the mRNA and protein expressions of TNF-α as well as mRNA expression of IL-1β.Caspase 3 protein expression significantly lowered in blood vessels in DEX group compared with LPS group.DEX had no obvious effect on lipopolysaccharide-induced vascular AMPK and CaSR mRNA or protein expressions.Conclusion DEX obviously deceased the expressions of inflammation-related proteins, suggesting that DEX has anti-inflammatory effects.

Objective To optimize the formula of konjac glucomannan-paeonol matrix tablets. Methods The formula of paeonol matrix tablets was optimized by the orthogonal design with the accumulative release rate in vitro as index, with the viscosity of konjac glucomannan ( KGM) , the amount of KGM and lactose as influence factors. Results The optimized formula was as follows:the viscosity of konjac glucomannan was 20 000 mPa·s, KGM 30%, lactose 20% and the release in vitro fit into the Higuchi equation. Conclusion The formula of the paeonol matrix tablets is reasonable and the tablets have well release effect in vitro.

Objective To explore the effect and mechanism of the bone morphologenetic protein 4 (BMP4)/Smad signaling pathway on the apoptosis of mouse primordial follicle oocytes .Methods Three-day-old Kunming mouse ovarine tissues were digested by the two-step enzymatic method to extract and purify oocytes .The cultured oocytes were divided into three groups: the normal culture medium (Con group), the medium with BMP4 (BMP4 group), and the medium with BMP4 and BMP4 inhibitor ( BMP4+inhibitor group ) .TUNEL was used to examine the effects of BMP 4 on the survival of the primordial follicle oocyte;Immunohistochemical staining and Real-time quantitative PCR were performed to investigate the expressions of p-Smad1/5/8, sohlh2, c-kit and foxo3a;siRNA interference, sohlh2 plasmid transfection and LY294002 treatments were performed to explore the mechanism of the BMP 4/Smad signaling pathway on the apoptosis of oocytes . Results TUNEL results demonstrated that the ratio of apoptotic oocytes in BMP 4 group was significantly lower than that in the Con group ( P <0.05 ) and the BMP4 +inhibitor group ( P <0.05 ); BMP4 significantly promoted the nuclear translocation of Smad and inhibited the nuclear translocation of foxo 3a, the mRNA and protein levels of sohlh2 and c-kit remarkably increased in BMP4 group.The effect of BMP4 on the oocyte survival was significantly repressed after sohlh 2 siRNA transfection.Sohlh2 overexpression up-regulated the expression of p-foxo3a, and this activity was abolished by LY 294002.Conclusion BMP4/Smad signaling pathway may inhibit primordial follicle oocyte apoptosis , via up-regulation of the expression of sohlh2 and c-kit, and then down-regulation of the nuclear translocation of foxo 3a.

AIM To investigate the anti oxidation activities of new systhesized nitroxides in liver, liver mitochondria and RBC from rats and in egg phospholipid. METHODS The homogenates of liver, liver mitochondria from rats and the suspensions of egg phospholipid were used to determine malondialdehyde (MDA) formation induced by Fe 2+ Vit C system using TBA colorimetric method. H 2O 2 caused hemolysis was measured spectrometrically. Superoxide anion was assayed spectrometrically. RESULTS Nitroxides A, B with one active group (NO?) could inhibit MDA generation caused by ?OH generation system significantly, antagonized hemolysis induced by H 2O 2, but did not affect O ? 2 formation; Nitroxide C with two active group (NO?) possessed similar potent anti lipoperoxidative activities,IC 50

Objective:To clone the human cell death-inducing DFF45-like effectors 3(CIDE3) full length cDNA for construction the eukaryotic expression vector pcDNA3.l(+)-CIDE3 and to study its bioactivity.Methods:① Total RNA was extracted from human white adipose tissues and the desired cDNA fragment was obtained by RT-PCR.After the fragment had being inserted into an eukaryotic expression vector pcDNA3.l(+),the resulted recombinant plasmid pcDNA3.l(+)-CIDE3 was transformed into DH5?.The positive clone was selected and confirmed to contain full length of CIDE3 cDNA by agarose gel and DNA sequence analysis.②After the pcDNA3.l(+)-CIDE3 plasmid was transfected into HeLa cells under lipofectamine mediation,the effect of target gene expression on growth of HeLa cells was analysed by TUNEL staining. Results:① The recombinant eukaryotic expression plasmid pcDNA3.l(+)-CIDE3 was constructed successfully and the sequence of CIDE3 was consistent with that of genebank.②After transfected pcDNA3.l(+)-CIDE3,HeLa cells presented distinguished apoptosis(about 15%),compared with that of transfected plasmid pcDNA3.l(+)(

Objective To investigate the parental origin for a rare case of complete hydatidiform mole and coexisting fetus and to discuss its diagnosis and differential diagnosis.Methods Tissues from the fetus,mole and placenta were collected and pathology analysis and chromosome analysis were done.The DNA from the fetus,mole and parents' peripheral blood leukocytes was amplified with five short tandem repeat (STR) markers (D4S2460,D18S488,D21S2039,DXS1205 and DYS219) at the same time to confirm the parental source of the hydatidiform.Results (1) Casereport:A 27-year-old woman,gravida 1,para 0,was found high risk for neural tube defects at 20 weeks of gestation.At 24+5 weeks of gestation,ultrasound examination demonstrated a normal fetus,a normal placenta and a huge mass with a multicystic appearance attached to the placenta with an obvious demarcation.The fetus died at 26 weeks of gestation.Serum human chorionic gonadotropin-β(β -hCG) level decreased obviously during the first two weeks after artificial induction,but elevated at the third week,and β-hCG titers fell to normal after 2 courses of chemotherapy.Fetus autopsy showed no structure abnormality.Histopathologic examination of the hydatidiform showed swelling of chorionic villi with hyperplasia of the trophoblast and formation of central cisterns suggesting of a twin pregnancy consisting of a complete hydatidiform mole and coexisting fetus.(2) Genetic analysis:The karyotype analysis of the normal placental villi was 46,XY; the cell cultures of fetal cartilage tissue and hydatidiform were failed.STR analysis showed that the fetus was diploid from biparental source;the mole was androgenetic source.And the mole had locus both from Y and X chromosome of the father,so it was heterozygous.It was suggested that this case was derived from one single oocyte fertilized with three spermatozoas.Conclusions STR analysis could be used to confirm the diagnosis of complete hydatidiform mole and coexisting fetus and to find the pathogenetic rnechanism.

Objective To investigate the effects of circumferential pulmonary vein isolation (CPVI) on atrial effective refractory period (ERP) in patients with paroxysmal atrial fibrillation.Methods 30 patients who underwent radiofrequency catheter ablation for paroxysmal AF were enrolled in this study.Using FAM mode,the RA and LA anatomical models were achieved in the CARTO 3 system.SVC,MRA,RAA,LA-A,LA-R,LA-P,LAA,LSPV,LIPV,RSPV,RIPV,CSp,CSd,were respectively located in the RA or LA anatomical model.Before and after CPVI,ERPs were measured in different locations of the atrium using programmed stimulation.The ERPs of the RA (SVC,MRA,RAA,CSp),LA (LA-A,LA-R,LA-P,LAA,CSd),PVs (LSPV,RSPV,LIPV,RIPV) were compared.Bilateral CPVIs were completed in all patients,and PV-LA bidirectional conduction block was achieved.The changes of electrophysiological characteristics of atrium before and after CPVI were observed.Results (1) ERP at different locations in the atrium before CPVI:Comparisons of ERPs at different locations of atrium:RAA had the minimal ERPs[(197.4 ± 28.6) ms (P ＜ 0.01);followed by PVs measuring,respectively,LSPV (213.0 ± 47.5) ms,LIPV (208.9 ± 45.9) ms,RSPV (209.3 ± 43.6) ms,RIPV (213.5 ± 48.1) ms and LAA (218.1 ± 27.7) ms.Comparisons of ERPs in RA,LA,and PVs showed:PVs had the lowest ERPs (211.2 ± 35.2) ms versus RA ERP (227.0 ± 23.7) ms versus LA ERP (241.0 ± 21.5) ms (P ＜ 0.05).(2) Comparisons of ERPs before and after CPVI:Comparisons of ERPs at different locations of atrium showed:RAA [(197.4 ± 28.6) ms vs.(208.6 ± 32.2) ms,P=0.003],CSp [(234.7 ± 29.1) ms vs.(246.9 ± 29.7) ms,P=0.007],LA-R [(242.9 ± 28.9) ms vs.(258.3 ± 26.9) ms,P=0.003],LA-P [(252.2 ± 28.5) ms vs.(261.1 ± 30.2) ms,P=0.039]and CSd [(238.6 ± 28.3) ms vs.(250.3 ± 23.6) ms,P =0.009].ERPs were found statistically prolonged at all different locations after CPVI.Comparisons of ERPs at RAand LA after CPVI showed:RA [(227.0 ± 23.7) ms vs.(235.9 ± 21.7)ms,P=0.002]and LA [(241.0 ± 21.5) ms vs.(249.7 ± 19.9) ms,P =0.001],which were statistically increased after CPVI.(3) A total of 90 episodes of atrial arrhythmias were induced before CPVI which were found at RAA (n =17),LAA (n =12),and PVs (n =36).After CPVI,8 episodes of atrial arrhythmias were induced which were found at,RAA (n =4),LAA (n =3),and SVC (n =1).Conclusions (1) Compared with other parts of atrium,ERPs at PVs,LAA and RAA are significantly shorter in patients with paroxysmal AF.At PVs,LAA and RAA,atrial arrhythmias are easily to be induce by programmed stimulation.(2) In patients with paroxysmal Af:PVs has the shortest ERPsfollowed by RAs whereas LA ERPs is the longest.There is a large ERP gradient change between PVs and LA.(3) The ERPs at RAs,LAs,As,and LA-PV are prolonged after CPVI.(4) Atrial arrhythmia is less likely to be induced after CPVI.

Objective To investigate the diagnostic potential of magnifying narrow-band imaging endoscopy (NBI-ME) for different intrapapillary capillary loop (IPCL) for the diagnosis of esophageal lesion.Methods Patients with abnormal esophageal mucosa found by white light gastroscopy in digestive endoscopy center,Chinese PLA General Hospital during the period of November 2009 to November 2010 were enrolled in this study.IPCL was observed and divided into different types by NBI-ME.Histopathology of biopsy or endoscopic submucosal dissection (ESD) specimens was evaluated and used as the gold standard to evaluate the diagnostic value of NBI-ME for IPCL.Results A total of 146 lesions from 145 subjects with esophageal mucosa abnormal were collected. Among them, 88 were pathology-proven inflammation,5 were pathology-proven esophageal cancers,20 were pathology-proven low intraepithelial neoplasia (LIN) and 33 were pathology-proven high intraepithelial neoplasia (HIN) detected with NBI-ME.By a per-lesion analysis,the accuracy of inflammation and cancer were 100％ (88/88) and 7/7.For the sensitivity,specificity,accuracy,positive predictive value,negative predictive value,positive likelihood ratio,negative likelihood ratio of LIN and HIN were 7/10,69.8％ ( 30/43 ),69.8％ ( 37/53 ),35.0％ (7/20),90.9％ (30/33),12.5％ (70/559),2.3％ (30/1290) and 87.1％ (27/31),72.7％ ( 16/22),81.1％ ( 43/53 ),81.8％ ( 27/33 ),80.0％ ( 16/20 ),634.1％ ( 837/132 ) and 35.2％ ( 124/352 ),respectively.Conclusions NBI-ME can classify the different esophageal IPCL.Higher diagnostic accuracy of IPCL indicates the feasibility of NBI-ME for the efficacious diagnosis of esophageal inflammation and cancer.There is the higher diagnostic accuracy of HIN than LIN.

There are some shortcomings of animal experiments applied in chemical toxicity testing, such as long period, large cost, species differences and dose differences, which limit the use of animal experiments' results in predicting human toxicity.Accordingly, we established a toxicity testing alternative screening system in line with the toxicity endpoints which required in chemical safety evaluation and risk assessment (genotoxicity, carcinogenicity, reproductive toxicity, acute toxicity and general toxicity) based on 3R principles (replacement, reduction, refinement) for animal experiments.This system covers most of the endpoints of toxicity assessment, and molecular biology technology was also applied to integrate the toxicity test, as well as some operation was optimized in order to shorten the experimental period, reduce experimental costs, improve animal welfare.Furthermore, the results from the screening system have higher clinical relevance because it is based on the toxicity mechanisms.