Objective: To establish an extraction and purification process line synchronization suitable for industrial production of glycyrrhizin acid and liquiritin. Methods: The extraction rates of glycyrrhizic acid and liquiritin were as indexes, orthogonal test was performed to determine the optimum conditions; The retention rate of glycyrrhizic acid and liquiritin and impurity removal rates were as indexes, the best ultrafiltration process parameters were optimized by orthogonal test. Results: The optimum extraction conditions were as follows: 0.75% ammonia water (24 times), extracted for three times, each time under 60 min, average rates of glycyrrhizic acid and liquiritin were 98.3% and 72.3%; The best ultrafiltration process parameters: 10 nm inorganic ceramic membrane, pressure of 0.12 MPa, temperature of 25℃. The average retention rates of glycyrrhizic acid and liquiritin were 99.3% and 98.9%, and an average removal rate of impurity was 23.3%. Conclusion: The experiment adopted the joint application of inorganic ceramic membrane ultrafiltration technology and ammonia extraction process, and it has realized the glycyrrhizic acid and licorice glycosides extraction and purification of synchronization; The process has low production cost and good safety, and is suitable for industrial application.

Objective To investigate the risk factors and prognosis on the post-resuscitation multiple organ dysfunction syndrome(MODS) after successful cardiopulmonary resuscitation(CPR).Methods Clinical data of 53 patients who were suffered with cardiac arrest(CA) and undergone successful CPR with return of spontaneous circulation(ROSC) were analyzed. Results There were 48 patients accompanied with MODS(90.6%),and among them,35 patients died in hospital(66.0%).All the 10 patients with CPR interval ≥6min were accompanied with MODS and they died in hospital.There were 43 patients who underwent CA immediately and of them,38 patients were accompanied with MODS.The incidence and mortality of MODS in the patients with CA-ROSC interval 0.05).Conclusion The risk factors such as ROSC interval ≥6 min,AC-ROSC interval ≥10min and the SIRS after ROSC are significantly associated with the incidence of MODS.The organic function of the patients should be evaluated promptly.

Objective To investigate the effects of methylprednisolone on the release of cytokines in patients after successful cardiopulmonary resuscitation(CPR).Method Thirty patients after CPR with restoration of spontaneous circulation(ROSC)were randomly divided into two groups:group A(methylprednisolone group,n=14)and group B(control group,n=16)during the period from May 2005 through May 2007.The patients of group were treated with methylprednisolone 3 mg/kg by intravenously twice a day after ROSC.The levels of serum tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),interleukin-6(IL-6),interleukin-8(IL-8),interleukin-10 (IL-10)were measured by enzyme-linked immunosorbent assay(ELISA)before CPR,and 24,48,72 hours and 7 days after ROSC.The data were analyzed studentis t test and chi-gquare test.A P value less than 0.05 indicated significant difference.Results There was on significant difference in the mean time from cardiac arrest to return of spontaneous circulation,and the levels of serum cytokines between the two groups before CPR(P＞0.05).In comparisorl with group B,the levels of serum TNF-α,IL-1β,IL-6,IL-8 decreased markedly at 24 and 48 hour after ROSC in group A(P＜0.05-0.01),and the levels of serum IL-8 decreased markedly at 72 hours after ROSC in groupA(P＜0.05).Thelevels of serum TNF-α,IL-1β,IL-6,IL-8 were not of significant differences between the two groups at 7 hys after ROSC(P＞0.05).There was no significant difference in the levels of serum IL-10between the two groups at different time points after ROSC(P＞0.05).Conclusions Methylprednisolone plays a role of preventive effects on patients with ROSC after PCR through decreasing the levels of serum TNF-α,IL-1β,IL-6,IL-8.

Objective To evaluate the effects of various polysorbates(PS)on the stability of different types of monoclonal antibody(mAb)drugs.Methods Three types of monoclonal antibodies mAbA(IgG1 proantibody drug),mAbB(IgG1 mAb)and mAbC(IgG1 mAb with Fc N297A mutation)were used as model proteins,and different kinds or contents of PS were added into the mAb formulations respectively to investigate the influencing factors.The effects of PS on the stability of mAb drugs were evaluated comprehensively by detecting the changes of quality attributes,such as protein aggregates and insoluble particles.Results PS20 and PS80 showed no significant difference in inhibiting the formation of aggregates and charge variants in the three mAbs(P>0.05),while the addition of PS80 in mAbB and PS20 in mAbC significantly inhibited the increase of insoluble particles respectively(P<0.05);The content of PS20 showed a significant effect on the detection indexes of charge variants and insoluble particles in mAbC(P<0.05).Conclusion Different types of mAbs have different sensitivities to various kinds and contents of PS.Therefore,when designing the formulation of mAbs,it is necessary to select appropriate kinds and contents of PS to further improve the stability of mAb drugs.

OBJECTIVETo detect the relationship between chromosomal anomalies and the pathogenesis, development and prognosis of ovarian carcinoma.

METHODSThirty-six specimens of ovarian carcinoma (n=12), ovarian benign tumor (n=12), and normal ovary (n=12) were examined by fluorescence in situ hybridization (FISH).

RESULTSTwelve cases of mutations, including trisomy 8, monosomy 8 or tetraploid 8 chromosomal anomalies, were found in the group of ovarian carcinoma, making up 100% (12/12). Three cases of trisomy 8 chromosomal anomalies were found in the group of ovarian benign tumor, accounting for 25% (3/12). No anomaly was found in the normal group. There were significant differences between the three groups, P<0.001.

CONCLUSIONThe above anomalies of chromosome 8 are significantly associated with the pathogenesis and development of ovarian carcinoma. The anomalies may occur in the early stage of the carcinoma, and may be significantly associated with the pathological differentiation and clinical stage of the case.

Adolescent ; Adult ; Aged ; Aneuploidy ; Chromosome Aberrations ; Chromosomes, Human, Pair 8 ; genetics ; Female ; Humans ; In Situ Hybridization, Fluorescence ; methods ; Middle Aged ; Monosomy ; Ovarian Neoplasms ; genetics ; Trisomy

To study the mechanism of thrombogenesis and search new anti-thrombotic agent, the cDNA for human vWF A1 domain was high-level expressed in E. coli and recombinant protein of vWF A1 with biologic activity was obtained. The gene encoding A1 domain was amplified by PCR from plasmid containing full length cDNA of human vWF. After confirming by DNA sequencing analysis, the recombinant expression plasmid pQE31-vWF/A1 was constructed and introduced into E. coli M15 strain, then induced by IPTG; the expressed protein was purified with Ni-NTA agarose, identified by Western blotting. The results showed that the 854 bp DNA fragment was obtained by PCR from the plasmid containing full length cDNA for human vWF and its sequence was identical to the published sequence. High level expression of A1 protein was yielded after 5 hour-induction, which amounted to 30% of total bacteria protein in inclusion body. Western blot demonstrated it possessed good antigenicity and high specificity. It is concluded that cDNA for vWF/A1 had been cloned successfully, high level expression of A1 protein was achieved in E. oli. This study will provide a basis for the further clinical and basic research on the role of vWF in thrombosis and hemostasis.
Blotting, Western ; Cloning, Molecular ; DNA, Complementary ; chemistry ; Escherichia coli ; genetics ; Humans ; Polymerase Chain Reaction ; Recombinant Proteins ; analysis ; von Willebrand Factor ; analysis ; biosynthesis ; genetics

The interaction among collagen, von Willebrand factor (vWF) and glycoprotein Ib axis is the first step in hemostasis and thrombosis, especially under high shear condition. To develop a new remedy of anti-thrombosis, mRNA from endothelial cells was extracted, and reverse transcription PCR was adopted to amplify DNA of interest. After sequencing, recombinant expression vector was constructed. The amplified DNA fragment of vWF domain A3 was inserted into expression vector with 6 x his taq, pET20b(+), the recombinant was transformed into E coli (strain DE3) and induced by IPTG. Recombinant vWF-A3 was designated as a recombinant fragment comprising residues 918 - 1114 of mature vWF subunit. It was purified through Ni-NTA resin column and refolded in Tris buffer containing GSH and GSSG. The results showed that rvWF-A3 was expressed successfully in E coli (strain DE3), accounting for 46% of total bacterial protein with its purity of over 95%. It was identified that rvWF-A3 is capable to bind collagen and inhibit the wild vWF binding to collagen by competition. It is concluded that rvWF-A3 fragment might be an effective antithrombotic agent for preventing arterial thrombosis.
Cloning, Molecular ; Collagen ; metabolism ; Escherichia coli ; genetics ; Humans ; Protein Structure, Tertiary ; Recombinant Proteins ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Sequence Analysis, DNA ; von Willebrand Factor ; chemistry ; genetics ; metabolism

OBJECTIVETo explore the incidence and angiographic features of exercise-induced ST-segment elevation in patients without prior myocardial infarction.

METHODSExercise-induced ST-segment elevation occurred in 15 out of 4601 consecutive patients without prior myocardial infarction underwent treadmill exercise testing during a 2-year period. The coronary angiographic features of the 15 patients (13 males, aged between 40 - 75 years) were analyzed.

RESULTSCoronary angiography revealed one hemodynamically relevant stenotic vessel in 6 patients, two hemodynamically relevant stenotic vessels in 6 patients, three hemodynamically relevant stenotic vessels in 3 patients. Left anterior descending (LAD) coronary artery was affected in 12 patients. Left main coronary artery (LMCA) stenosis was evidenced in 1 patient and right coronary artery stenosis in 7 patients. Severe (90% - 100%) occlusions were visualized in 8 out of 13 patients with LAD or LMCA lesions. Elevated ST-segment leads were consistent with the ischemic area where the blood supply of myocardium was affected by diseased vessels.

CONCLUSIONSThe incidence of exercise induced ST-segment elevation in patients without prior myocardial infarction is very low and mostly due to severe fixed coronary artery stenosis, especially in LAD. The location of ischemic myocardium can be suggested by ST-segment elevation leads during exercise.

Adult ; Aged ; Coronary Angiography ; Coronary Artery Disease ; diagnosis ; physiopathology ; Electrocardiography ; Exercise Test ; Female ; Humans ; Male ; Middle Aged

OBJECTIVETo study the inhibitory effects of Panax Notoginseng Saponins(PNS) on apoptosis induced by hypoxia/hypoglycemia and reoxygenation in cultured rat hippocampal neurons.

METHODApoptosis were measured by flow cytometry, intracellular free calcium concentration([Ca2+]i) was measured with confocal laser scanning microscopy, morphological changes and neuronal necrosis were observed with fluorescence microscope, and meanwhile the leakage of lactic dehydrogenase(LDH) was measured.

RESULTHypoxia/hypoglycemia cultures for 5 hours and reoxygenation induced neuronal apoptosis and necrosis, and significantly increased neuronal [Ca2+]i and the leakage of LDH. The effects were increased with the extending time of reoxygenation. PNS has could significantly decrease the percentage of neuronal apoptosis and necrosis, and reduce neuronal [Ca2+]i and the leakage of LDH.

CONCLUSIONPNS has inhibitory effect on neuronal apoptosis. This effect might be related to its effect of decreasing intracellular free calcium concentration.

Animals ; Apoptosis ; drug effects ; Calcium ; metabolism ; Cell Hypoxia ; Cells, Cultured ; Fetus ; Ginsenosides ; isolation & purification ; pharmacology ; Hippocampus ; cytology ; Hypoglycemia ; pathology ; L-Lactate Dehydrogenase ; metabolism ; Neurons ; cytology ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar

OBJECTIVETo explore the effects of prenatal exposure to stress and lead on spatial learning and memory development in rats.

METHODSAll 32 Sprague-Dawley (SD) pregnant rats were divided randomly into 4 groups, 8 per group in line with the Random Number Table. The four groups were: no maternal stress, no Pb exposure (NS/C); non-maternal stress, Pb exposure (NS/L), maternal stress, no Pb exposure (S/C), and maternal stress plus Pb exposure (S/L). The spatial learning and memory ability, the serum corticosterone level both pre and post-testing of 30-day old offsprings, and lead concentration in hippocampus were tested by means of Morris Water Maze, radioimmunoassay and Inductively Coupled Plasma Mass Spectrometry (ICP-MS).

RESULTSThe residence time of male and female in NS/L was (16.08+/-3.41) s, (15.72+/-3.33) s, which were significantly shorter than NS/L (25.42+/-4.76) s, (24.55+/-4.43) s and S/C (20.96+/-3.45) s, (20.65+/-2.98) s, and significant difference was observed in the joint exposure effect (F=5.478, P<0.05). The effect of the joint exposure was significant on post-testing serum corticosterone. The hippocampus lead concentrations of NS/L and S/L were (0.4378+/-0.1041) microg/g and (0.4679+/-0.1243) microg/g without significant differences (F=0.298, P>0.05).

CONCLUSIONPrenatal joint exposure to restraint stress and lead might increase the effects of single exposure on the spatial learning and memory ability and serum corticosterone level of offsprings, and the joint influence on corticosterone level might be one of the reasons of further impairment of learning and memory.

Animals ; Animals, Newborn ; Environmental Exposure ; Female ; Lead ; toxicity ; Male ; Maze Learning ; drug effects ; Memory ; drug effects ; Pregnancy ; Prenatal Exposure Delayed Effects ; Rats ; Rats, Sprague-Dawley