Objective: To study the in vivo metabolic pathways of liquiritin (LQ) in rats. Methods: An HPLC-QTRAP-MS method was established and applied to identify the metabolites of LQ in bile, urine, feces, and plasma after ig administration of LQ (300 mg/kg) to rats. Results: A total of nine metabolites were found in rats. The major metabolic pathway of LQ was deglucosidation to liquiritigenin (LG) and dehydration, glucuronidation, and sulfation of LG. Conclusion: LQ undergoes extensive phases I and II metabolism in rats. The major metabolites of LQ are LG and its glucuronides and sulfates.

Objective: A simple, sensitive, and rapid LC-MS/MS method has been established and validated for the determination of liquiritigenin (LG) in rat plasma. Methods: Naringenin was chosen as internal standard (IS). LG and IS were separated on a Diamonsil C18 analytical column with a mobile phase of methanol-10% methanol in water containing 0.5 mmol/L ammonium formate and 0.2% formic acid (55:45) at the isocratic flow rate of 0.6 mL/min for 10 min. The multiple reaction monitoring (MRM) was performed on a mass spectrometer in the negative ion mode with electro-spray ionization (ESI) source and the transition from precursor ion to product ion was m/z 255.0→119.0 for LG and m/z 271.0→151.0 for IS, respectively. Results: The linearity was acceptable in the range of 5-5000 ng/mL (r = 0.9973). The inter-day and intra-day accuracies were in the ranges of -0.09%-3.25% and -5.02%-9.21%, respectively. The precision was in the ranges of 3.60%-12.4% and 0.909%-6.89%, respectively. LG was stable in the course of analysis and storage. Conclusion: The LC-MS/MS method was successfully applied to the pharmacokinetic study for the first time in rats after ig and iv administration of liquiritin (LQ), a glycoside of LG, at pharmacologically effective levels.

Objective To investigate the relationship between subclinical hypothyroidism (SCH) and diabetic retinopathy (DR) in patients with type 2 diabetes mellitus (T2DM).Methods A total of 792 patients of T2DM were enrolled in the study.There were 448 males and 344 females,with an average age of (54.13 ± 13.06)years.The average duration of diabetes was (8.03 4±6.70) years.The patients were grouped according to the degree of DR and thyroid function.Among them,483 patients (61.0％) were no DR,240 patients (30.3％) were mild DR,69 patients (8.7％) were severe DR.725 patients (91.5％) were normal thyroid function,67 patients (8.5％) were SCH.The prevalence of SCH among no DR group,mild DR group and severe DR group was compared.And the prevalence of DR between normal thyroid function group and SCH group was compared.Logistic regression analysis was used to estimate the association between SCH and DR.Results No significant differences among the three groups (no DR group,mild DR group,severe DR group) were found in the prevalence of SCH (x2=1.823,P=0.402).There were no significant differences in the incidences of DR between normal thyroid function group and SCH group (x2=1.618,P=0.239).Logistic regression analysis demonstrated that SCH was not significant associated with DR [mild DR:odds ratio (OR)=1.361,95％ confidence interval (CI)=0.773-2.399,P=0.286;severe DR:OR=1.326,95％CI=0.520-3.384,P=0.555;DR:OR=1.353,95％ CI=0.798-2.294,P=0.261).Conclusion SCH is not significant associated with DR in patients with T2DM.

Objective To observe the biological activities of human doppel(Dpl) protein transiently expressed and Dpl-like protein PrP?32-121 on a human neuroblastoma cell line SH-SY5Y.Methods Recombinant mammalian expression plasmids containing human PRND gene and truncated PrP?32-121 fragment were generated by PCR.The expression and location of Dpl and PrP?32-121 post-transfection were observed by IFA.The cytotoxicity was measured by MTT analysis.Cellular apoptosis was investigated by flow cytometry and Western blot.Results Both Dpl and PrP?32-121 protein were expressed and mainly located on the cell membrane.Remarkable cytotoxicity was detected on SH-SY5Y cells after 24 h transfection.Meanwhile,more Annexin V/PI positively-stained cells as well as lower levels of cellular pro-caspase-3 and Bel-2 were detected in the cells receiving Dpl and PrP?32-121 expressing plasmids.Conclusion Dpl protein transiently expressed and PrP?32-121 can lead to the similar neural cytotoxicity,probably triggering the cell apoptosis program.

Objective To observe the biological activities of human doppel (Dpl) protein transiently expressed and Dpl-like protein PrPΔ32-121 on a human neuroblastoma cell line SH-SY5Y. Methods Recombinant mammalian expression plasmids containing human PRND gene and truncated PrPΔ32-121 fragment were generated by PCR. The expression and location of Dpl and PrPΔ32-121 post-transfection were observed by IFA. The cytotoxicity was measured by MTT analysis. Cellular apoptosis was investigated by flow cytometry and Western blot. Results Both Dpl and PrPΔ32-121 protein were expressed and mainly located on the cell membrane. Remarkable cytotoxicity was detected on SH-SY5Y cells after 24 h transfection. Meanwhile, more Annexin V/PI positively-stained cells as well as lower levels of cellular pro-caspase-3 and Bel-2 were detected in the cells receiving Dpl and PrPΔ32-121 expressing plasmids. Conclusion Dpl protein transiently expressed and PrPΔ32-121 can lead to the similar neural cytotoxicity, probably triggering the cell apoptosis program.

Objective To elucidate the effects of different modes of partial parenteral nutrition (PPN)on immunological function of T-lymphocyte in non small cell lung cancer(NSCLC)patients during chemotherapy.Methods Ninety-three patients with non-small cell lung cancer were randomly divided into three groups:the control group(30 patients),the low dose of PPN(32 patients) and the high dose of NNP(31 patients).Exactly the same chemotherapy was applied to each of three groups.During chemotherapy,three groups were supplied the same diet,the control group received conventional treatment;the low dose group and the high dose group received additional parenteral nutritional support besides diet.The low dose group was given 250 ml 9-AA daily and the high dose group was given 500 ml 9-AA daily.The T lymphocyte subsets CD3~+,CD3~++CD4~+ ,CD3~++CD8~+ and cells were detected respectively before and after chemotherapy.Results In all of the three groups,the percentage of NK cells,CD3~+ and CD3~++CD4~+ cells were decreased significantly before and after chemotherapy(all P＜0.05),In the control and low dose groups,NK cells changed more significantly after chemotherapy(P＜0.01).The percentages of CD3~+,CD3~++CD4~+,CD3~++ CD4~+/CD3~++CD8~+ of the low dose group and high dose group were higher than those of the control group before and after chemotherapy(all P＜0.05),the percentage of CD3~++CD8~+,CD3~++CD4~+/ CD3~++CD8~+ of the low dose group and hight dose group did not change notably(all P＞0.05). Conclusions The chemotherapy on patients with NSCLC will possibly cause malnutrition and immunosuppression.The benefits of giving 9-AA to NSCLC patients who were applying PPN and undergoing chemotherapy may include antagonizing immunological function aggravation,improving nutrition status and improving immunological functions of the T lymphocytes during chemotherapy.

BACKGROUND: As a micro-injurying and reduplicative treatment of osteoarthritis,the arthroscopic debridement has got the affirmation of numerous scholars. But as one of the standard procedures in artnroscopic debridement,synovectomy is called in question recently.OBJECTIVE: To explore the applied value of synovectomy in the arthroscopic debridement of osteoarthritis.DESIGN: Retrospective controlled analysis.SETTING: Department of Orthopaedics, Fushun Central Hospital.PARTICIPANTS: Sixty-five patients received synovectomy in the arthroscopic debridement of knee joint osteoarthritis in the Department of Orthopaedics,Fushun Central Hospital from February 1997 to December 2000.Thirty-two among them,with complete data and over 1 year's followup,were taken for synovectomy group. Forty-eight patients received the arthroscopic debridement of knee joint osteoarthritis without synovectomy in the Department of Orthopaedics,Fushun Central Hospital from January 2001 to November 2003.Thirty among them,with complete data and over 1 year's follow-up,were taken for control group.METHODS: Synovectomy was taken as the factor of intervention in the operation to perform grouping. The analysis of curative effect was performed to control with joint douching,corpus liberum removal,osteophyma cleaning,meniscus fitting,cartilage gouging,synovectomy and without synovectomy or part synovectomy. Lysholm evaluation standard of knee joint osteoarthritis was used to the knee joint functional evaluation beween preoperation and postoperative 1 year in two groups. And operative time,postoperative draining quantity,postoperative 7-day visual analog score,postoperative 1-year Lysholm score of knee joint, were recorded.MAIN OUTCOME MEASURES: Preoperative and postoperative 1-year Lysholm score of knee joint,operative time,postoperative draining quantity,postoperative 7-day visual analog score.RESULTS: Sixty-two patients were included and all of them entered the result analysis.The preoperative patients in two groups were comparable with each other and the differences of Lysholm score were not significant (t=0.127,P=0.899).The operative time was longer in synovectomy group than in control group,the differences were significant (t=9.547,P ＜ 0.001)and the postoperative draining quantity was more in synovectomy group.The postoperative visual analog score was bigger in synovectomy group than in control group and the differences were significant [the scores of synovectomy and control groups were respectively (4.6±1.1),(2.8±1.4),t=6.206,P ＜ 0.001].The differences of knee joint score in 1-year follow-up were not significant [the scores of synovectomy and control groups were respectively (77.6±11.9),(79.0± 10.3),t=0.562,P=0.576].CONCLUSION: Synovectomy can not increase the curative effect in the near future in the arthroscopic debridement of osteoarthritis. On the contrary,the operative time was longer,the operative wound was larger and postoperative reaction was more serious. It should not be used in the debridement generally.

10.3969/j.issn.2095-4344.2013.24.016

Objective:This work aims to investigate diallyl disulfide (DADS) inhibition of cell migration and invasion in human colon cancer SW480 cells through the Rac1-ADF/cofilin1 pathway. Methods:The potential of cell migration and invasion was examined by scratch healing assay and transwell membrane assay. The expression of Rac1-ADF/cofilin1 pathway was detected by RT-PCR and Western blot. Results:After the SW480 cells were treated with 40 and 50 mg·L-1 of DADS for 24 h, the number of transmembrane cells through the Matrigel obviously decreased by 57.12%and 64.59%, respectively (P<0.05). After cell treatment for 48 h, the cell migration rates were 23.23%and 12.87%, which were significantly lower compared with the control group (75.86%;P<0.05). After the cells were treated with 45 mg·L-1 of DADS for 24 and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and destrin mRNA respectively decreased compared with the control group (P<0.05). However, no significant difference was observed in the expression of cofilin1 mRNA (P>0.05). After the treatment with 45 mg·L-1 of DADS for 6, 12, 24, and 48 h, the expression of Rac1, Rock1, PAK1, LIMK1, and Destrin proteins respectively decreased in a time-dependent manner compared with the control group (P<0.05). However, no significant differences were observed in the expression of the cofilin1 protein (P>0.05). Moreover, the expression of p-LIMK1 and p-cofilin1 notably decreased in a time-dependent manner (P<0.05). Conclusion:DADS inhibits cell migration and invasion, which is related to the down-regulation of Rac1, Rock1, PAK1, LIMK1, p-LIMK1, p-cofilin1, and destrin through the Rac1-ADF/cofilin1 pathway.

Objective To explore the relation between plasm neurotransmitters (Glutamic acid,GAA; γ-aminobutyric acid,GABA; 5-hydroxytryptamine,5-HT; and noradrenaline,NE) and depression in acute hemorrhagic stroke.Methods Objectives were screened from consecutive hospitalized patients with acute stroke.Fasting blood samples were taken on the day next to hospital admission,and neurotransmitters were examined by the liquid chromatography-high resolution mass spectrometry (LC-HRMS).The fourth edition of Diagnostic and Statistical Manual of Mental Disorders (DSM-Ⅳ) was used to diagnose depression at two weeks after onset of stroke.The modified Ranking Scale (mRS) was followed up at 1 year.Pearson test was used to analyse the correlation between serum concentration of neurotransmitters and the Hamilton Depression scale-17items (HAMD-17) score.Logistic regression was used to analyse the relation of serum concentration of neurotransmitters and depression and outcome of stroke.Results One hundred and eightyone patients were included in this study.GABA significantly decreased [6.1 (5.0-8.2) μg/L vs 8.1 (6.3-14.7) μg/L,P ＜ 0.05] in patients with depression in hemorrhagic stroke,and there was no significant difference in GAA,5-HT,or NE.GABA concentration was negatively correlated with HAMD-17 score (r =-0.131,P ＜ 0.05) ; while concentration of serum GABA rose by 1 μg/L,risk of depression in acute phase of hemorrhagic stroke was reduced by 5.6％ (OR 0.944,95％ CI 0.893-0.997).While concentration of serum GAA rose by 1 μg/L,risk of worse outcome at 1 year was raised by 0.1％,although a statistic level was on marginal status (OR 1.001,95％ CI 1.000-1.002).Conclusions In patients with depression in the acute phase of hemorrhagic stroke,there was a significant reduction in plasm GABA concentration.GABA may have a protective effect on depression in acute phase of hemorrhagic stroke.Increased concentrations of serum GAA may increase the risk of worse outcomes at 1 year after stroke.