Objective To develop a gas chromatography-mass spectrometry (GC-MS) detection method for quantification of the nonpolar amino acids in amniotic fluid of congenital malformation including alanine (Ala),leu cine (Leu),isoleucine (Ile),proline (Pro) and phenylalanine (Phe).To compare the different amino acids in amniotic fluid between pregnant women with congenital malformation and normal control and analyze the corresponding pathogenesis.Methods After precipitated protein of the 100 μL supernatant of amniotic fluid by methyl alcohol,the supernatant was dried by nitrogen.The extractions were treated with MSTFA for derivatization.Then gas chromatography-mass spectrometry was used to detect and analyze the amino acids.Results This method was proved to have good sensitivity,precision,accuracy and recoveries.Under the optimum testing conditions,the limit of detection (LOD) was 0.12-0.38 mg/L.The calibration curves showed good linearity over the investigated concentration range between 0.5 and 10 mg/L.The recoveries were 91.12％ to 104.41％.The relative standard deviation of intra and inter-day was 0.84％ to 9.33％.The developed method was applied to the quantification of 5 nonpolar amino acids in amniotic fluid of 17 pregnant women with congenital malformation and 13 normal control pregnant woman.The contents of leucine and isoleucine decreased in disease group compared to controls.The difference of the other three amino acids between the two groups had no statistical significance.Conclusions The validation results showed that the method was suitable for detection of the amino acids in amniotic fluid and having broad prospect of clinical application.Leucine may participate in the pathogenesis of congenital malformation.

Objective To investigate the changes in cerebral oxygen metabolism during liver transplantation in patients with liver cirrhoses.Methods Sixteen ASAⅢorⅣpatients with liver cirrhoses(14 male,2 female)aged 25-67 yrs,weighing 45-80 kg undergoing liver transplantation were studied.Radial artery was cannulated for direct BP monitoring and blood sampling.Swan-Ganz catheter was placed in pulmonary artery (PA)via right internal jugular vein(IJV)for cardiac output(CO)monitoring and sampling mixed venous blood. Left IJV was cannulated and the catheter was advanced cephalad until jugular bulb for blood sampling.Anesthesia was induced with midazolam,fentanyl,propefol and vecuronium and maintained with isoflurane inhalation and intermittentⅣboluses of fentanyl and vecuronium.The patients were mechanically ventilated after tracheal intubation.PaCO_2 was maintained between 30-45 mm Hg.Blood samples were taken from radial artery,pulmonary artery and jugular bulb simultaneously for blood gas analysis before operation(T_0,baseline),10 min before anhepatic phase(T_1)20 min after onset of anhepatic phase(T_2),30 min after graft reperfusion(T_3)and at the end of operation(T_4).Oxygen delivery(DO_2),oxygen consumption(VO_2),oxygen content of jugular bulb blood (CjvO_2),cerebral arterial-venous oxygen content differences(Ca-jvO_2)cerebral oxygen extraction ratio(CERO_2) and CBF/CMRO_2 were calculated.Results The mean duration of operation was(364?51)min and the mean intraoperative blood loss was(1340?430)ml.CO was significantly increased before anhepatic phase(T_1), during neohepatic phase(T_3)and at the end of operation(T_4)but decreased during anhepatc phase(T_2)as compared with the baseline value at T_0.Hb,CaO_2,Ca-jvO_2 and CERO_2 were all decreased while SjvO_2 and CBF/ CMRO_2 were increased during operation;DO_2,VO_2 and CjvO_2 were decreased during anhepatic phase;DO_2 was increased during other phases;VO_2 was increased at the end of operation as compared with the baseline(T_0)(P＜0.05 or 0.01).Conclusion There is no cerebral oxygen deficiency during liver transplantation in patients with liver cirrhoses.

Objective To investigate the changes in systemic and pulmonary hemodynamics in patients with liver cirrhosis and portopulmonary hypertension(PPH)during liver transplantation.Methods Eight patients with liver cirrhosis and PPH(5 male,3 female)aged 50-63 yr weighing 45-80 kg were included in PPH group. Another 8 liver-cirrhotic patients without PPH served as control group.The patients were premedicated with intramuscular phenobarbital 0.1 g and atropine 0.5 mg.Anesthesia was induced with midazolam 3-5 mg,fentanyl 0.15-0.2 mg,propefol 1 mg?kg~(-1) and vecuronium 0.1 mg?kg~(-1) and maintained with 0.5%-3% isoflurane inhalation and intermittent Ⅳ boluses of fentanyl and vecuronium.The patients were mechanically ventilated after tracheal intubation.P_(ET)CO_2 was maintained at 30-45 mm Hg.Right subclavian vein was cannulated for fluid and drug administration and blood transfusion.Radial artery was cannulated for BP monitoring.Swan-Ganz catheter was placed via right internal jugular vein.BP,CVP,MPAP,PAWP,CI,PVRI and SVRI were monitored and recorded before operation(baseline),during preanhepatic phase,at 3 and 30 min of anhepatic phase and 3,7, 15,60 min of neohepatic phase and at the end of operation.Results(1)The two groups were comparable with respect to fluid balance,the amount of vasoactive drugs used during anhepatic and neohepatic phase,duration of anhepatic phase and operation.(2)MPAP and PVRI were significantly higher before operation in PPH group than in control group.(3)CI,MPAP, PAWP and CVP were siguificanfly decreased during anhepatic phase as compared to the baseline values(before operation)in both groups and then gradually returned to and even exceeded the baseline values during neohepatic phase.(4)During neohepatic phase PVRI in PPH group was significantly increased as compared to the baseline value and was significantly higher than that in control group.Conclusion MPAP and PVRI are significantly increased during neohepatic phase in patients with PPH and need to be treated.

AlM: To study the clinical effect of the application of microscopic pterygium resection combined with different concentration of mitomycin C ( MMC) .
METHODS:A total of 110 cases of pterygium patients (120 eyes) were randomly divided into control group (58 eyes) and observation group (62 eyes) according to the odd and even number method. The control group adopted the pterygium resection combined 0. 3mg/mL MMC, and the observation group was given pterygium resection combined 0. 2mg/mL MMC. The cure rate and the recurrence rate, eyesight before and after the treatment, two groups of cornea and sclera wound healing situation, the incidence of postoperative complications were compared.
RESULTS: The cure rate and recurrence rate of the control group was 84. 5% and 15. 5% respectively, and the observation group was 93. 6% and 6. 5% respectively, the differences were statistically significant (P<0. 05). There were statistical differences of vision of the two groups before and after treatment (P<0. 05), and there were no statistical differences of the two groups between the two groups after treatment (P>0. 05). The cornea, sclera, wound healing time of the observation group were less than the control group, and there were statistical differences between the two groups ( P < 0. 05 ). The incidence of complications was 13. 8% in the control group and 3. 2% in observation group, with statistically significant difference (P<0. 05).
CONCLUSlON: The application effect of microscopic pterygium resection combined with MMC is remarkable, and the joint of 0. 2mg/mL concentration of MMC is more safe and effective, and is worth popularizing in clinical application.

Objective To summarize the experiences in using Bio-Gide and Bio-Oss for guided bone regener- ation in dental implantation.Methods In 28 cases of bone deficiency,Bio-Gide membranes were applied to cover alveolar defects filled with the Bio-Oss bone powder.In postoperative periodic follow-.up,the bone regeneration effect was observed by successive clinical and X-ray examination.Results 38 implants were inserted in the 28 patients and Bio-Gide membranes were used in the sites of the 38 implants.Alveolar bone defects were filled with new bone in 27 patients,1 implant loosed because of inflammation.37 implants had ideal osseointegration at stageⅡsurgery and were prosthetic rcconstructed successfully.No implant loosed during the observed period of 15 months to 4 years. Conclusion Bio-Gide and Bio-Oss have ideal effect of guided bone regeneration in dental implantation.

OBJECTIVETo observe the clinical effect of indwelling the anterior urethral stent in the treatment of anterior urethral stricture.

METHODSWe included 38 patients with anterior urethral stricture in the treatment group, and another 38 in the control, the former treated by indwelling the anterior urethral stent, and the latter by urethral dilatation. Then we analyzed the clinical results by comparing the Qmax, urinary hesitancy and numbers of urethral dilations between the two groups.

RESULTSCompared with the controls, the patients of the treatment group showed an obvious increase in Qmax, a significant decrease in the number of urethral dilatations, and a marked improvement of the quality of life. Six months after the stent removal, there were significantly more patients with Qmax > 15 ml/s in the treatment group than in the control (P < 0.05).

CONCLUSIONIndwelling the anterior urethral stent is a desirable option for the treatment of anterior urethral stricture, which is simple, safe, effective and reliable, and can be applied to general clinical practice.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Humans ; Male ; Middle Aged ; Stents ; Treatment Outcome ; Urethra ; surgery ; Urethral Stricture ; surgery

This study was aimed to investigate the expression and regulation mechanism of DNA-dependent protein kinase catalylic subunit (DNA-PKcs) in chronic myeloid leukemia (CML) and its role in blast crisis of CML. Expression of DNA-PKcs mRNA was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and DNA-PKcs protein by Western blot in 62 CML patients and K562, as compared to those of 23 normal individual controls. In 26 CML patients received allogeneic peripheral blood stem cell transplantation (allo-PBSCT) and 4 CML patients treated with imatinib, the expression of bcr-abl mRNA and DNA-PKcs protein was detected by RT-PCR and Western blot, respectively. After treatment with imatinib in mononuclear cell (MNC) of CML patients and K562 in vitro, expression of DNA-PKcs mRNA was detected by RT-PCR and DNA-PKcs protein level, tyrosine phosphorylation of bcr-abl fusion protein were detected by Western blot. The results showed that the expression of DNA-PKcs protein was significantly lower in CML and K562 than those in normal control (P<0.05). In 26 CML patients received allo-PBSCT and 4 CML patients treated with imatinib, the expression of DNA-PKcs protein was enhanced while the expression of bcr-abl mRNA decreased. After treatment of MNC of CML and K562 with imatinib in vitro, the expression of DNA-PKcs protein was enhanced while tyrosine phosphorylation of bcr-abl fusion protein decreased. It is concluded that the expression of DNA-PKcs protein is down-regulate by bcr-abl fusion gene, and the bcr-abl fusion gene down-regulate the expression of DNA-PKcs protein by post-transcriptional mechanism; the decrease of DNA-PKcs protein expression may be one of mechanisms underlying the acute transformation of CML.
Adult ; Aged ; Benzamides ; Bone Marrow Cells ; metabolism ; DNA-Activated Protein Kinase ; biosynthesis ; genetics ; Female ; Fusion Proteins, bcr-abl ; biosynthesis ; genetics ; Humans ; Imatinib Mesylate ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; therapy ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Piperazines ; therapeutic use ; Pyrimidines ; therapeutic use ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVETo observe the effects of astragalus membranacaus injection on the activity of the intestinal mucosal mast cells (IMMC) and inflammatory response after hemorrahagic shock-reperfusion in rats.

METHODThirty-two Wistar rats were randomly divided into four groups: normal group, model group, low dosage group, (treated with astragalus membranacaus 10 g kg(-1)) and high dosage group (treated with astragalus membranacaus 20 g kg(-1)). Models of hemorrhage shock for 60 minutes and reperfusion for 90 minutes were created. The animals were administrated 3 mL therapeutic solution before reperfusion. At the end of study, intestinal pathology, ultrastructure of IMMC, and expression of tryptase were observed. The levels of MDA, TNF-a, histamine, and SOD activity of intestinal were detected, and the number of IMMC was counted.

RESULTThe degranulation of IMMC was seen in model group and was attenuated by astragalus membranacaus treatment. Chiu's score of model group was higher than that of the other groups. Astragalus membranacaus could attenuate the up-regulation of the Chiu' s score, the levels of MDA and TNF-alpha, expression of tryptase, and the down-regulation of SOD activity and histamine concentration. The Chiu's score and MDA content were negatively, while SOD activity was positively correlated to the histamine concentration respectively in the four groups.

CONCLUSIONAstragalus membranacaus can reduce small intestine mucosal damage by inhibiting the activity of IMMC after hemorrhage shock reperfusion.

Animals ; Astragalus membranaceus ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Injections, Intravenous ; Intestinal Mucosa ; metabolism ; pathology ; Intestine, Small ; metabolism ; Male ; Malondialdehyde ; metabolism ; Mast Cells ; drug effects ; metabolism ; ultrastructure ; Random Allocation ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism ; pathology ; Shock, Hemorrhagic ; metabolism ; pathology ; Tryptases ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo investigate the expression and regulation mechanism of mismatch repair (MMR) genes in chronic myeloid leukemia (CML).

METHODSExpression of MMR genes hMSH2, hMSH3, hMSH6, hMLH1 and hPMS2 mRNAs in 62 CML patients and K562 cell line were detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Expression of bcr-abl mRNA and MMR genes mRNA were detected by RT-PCR in 26 CML patients with allogeneic peripheral blood stem cell transplantation (allo-PBSCT) and 4 CML patients on imatinib treatment. Expression of bcr-abl mRNA was detected by RT-PCR and tyrosine phosphorylation of BCR-ABL fusion protein by Western blot.

RESULTSExpression of hMSH2, hMSH3 and hMLH1 mRNA was significantly lower in CML and K562 cells than in normal control (P < 0.05). In 26 CML with allo-PBSCT and 4 CML patients on imatinib treatment, expressions of hMSH2, hMSH3 and hMLH1 mRNA was enhanced while expression of bcr-abl mRNA decreased. In CML MNC after imatinib treatment and in K562 cells, expression of hMSH2, hMSH3 and hMLH1 mRNA was enhanced while tyrosine phosphorylation of BCR-ABL fusion protein decreased.

CONCLUSIONExpressions of hMSH2, hMSH3 and hMLH1 mRNA were down-regulated by bcr-abl fusion gene.

Adult ; Aged ; Antineoplastic Agents ; pharmacology ; Benzamides ; DNA Mismatch Repair ; Female ; Fusion Proteins, bcr-abl ; genetics ; metabolism ; Gene Expression ; Gene Expression Regulation, Neoplastic ; Humans ; Imatinib Mesylate ; K562 Cells ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Male ; Middle Aged ; Piperazines ; pharmacology ; Pyrimidines ; pharmacology ; RNA, Messenger ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo establish a rapid, relatively quantitative method of detecting acetylated proteins.

METHODSThe proteins of Jurkat cells were acetylated by Trichostatin A (TSA) at different concentrations, then enriched and purified by anti-acetylated lysine antibodies affinity chromatography colum. The components eluted by acid were fixed on the microplate, the levels of acetylated proteins were tested by ELISA, and their components were identified by MALDI-TOF-TOF mass spectrometry. Also the above-mentioned methods were applied to the other three agents (gallic acid, emodin and monoacetylated emodin A).

RESULTSThat 4 × 10(5) Jurkat cells treated with 1 µmol/L TSA produced the optimal acetylated effect, up to 22 acetylated proteins were identified by MALDI-TOF-TOF, of them 15 were acetylated histones. The other three agents also induced acetylation, the relative values of acetylated proteins of Jurkat cells treated with 35.09 µmol/L and 17.54 µmol/L gallic acid were 4.3% and 14.2% respectively; those as of 28.7% and 11.5% treated with 1.47 µmol/L and 2.94 µmol/L emodin; those as of 22.0% and 3.6% treated with 152.91 µmol/L and 30.58 µmol/L monoacetylated emodin A.

CONCLUSIONThe method based on affinity chromatography colum may be useful for the detection of acetylated proteins, and could be used to screen agents which target to histone deacetylase.

Acetylation ; Chromatography, Affinity ; Histones ; analysis ; Humans ; Hydroxamic Acids ; pharmacology ; Jurkat Cells ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization