Research status of medicinal value of solanine from potato metabolite / 中国药理学通报

Potato skin may turn green, purple or sprout when stored improperly.At this time the content of its metabolite sola- nine increases significantly.Although solanine is toxie,it also has certain medicinal value.It has been found that solanine has effects like anti-cancer, anti-inflammation, detumescence, treatment of pulmonary hypertension, and diabetes.This paper searches the relevant literature for the research on the medicinal application of solanine,and systematically summarizes the solanine medicinal value.For a more effective development and utilization of the plant resources,this paper hopes to provide the theoretical and application basis for the development of natural medicine with high efficiency and small adverse reactions in the treatment of cancer, inflammation, tumescence, pulmonary hypertension and diabetes.

Bone marrow mesenchymal stem cell-derived exosomes improve hippocampal neuron injury in rats with depression via microRNA-1297/CTGF axis / 中华微生物学和免疫学杂志

Objective:To investigate the effect and mechanism of bone marrow mesenchymal stem cell (BMSC)-derived exosomal microRNA-1297 (miR-1297) on hippocampal neuron damage in depressed rats.Methods:BMSCs and BMSCs-derived exosomes were prepared and identified. Rats were first injected with corticosterone to establish the model of depression, and then injected with BMSCs-derived exosomes. Superoxide dismutase (SOD), malondialdehyde (MDA), lactate dehydrogenase (LDH), TNF-α and IL-1β in rat serum samples, hippocampal tissues and neurons were detected. Expression of miR-1297 in hippocampal tissues and neurons was detected by RT-qPCR. A rat hippocampal neuron injury model was established to investigate the role of BMSC-derived exosomes and miR-1297 in neuronal apoptosis and proliferation. The targeting relationship between miR-1297 and connective tissue growth factor (CTGF) was analyzed using dual luciferase reporter genes.Results:In the hippocampus of depressed rats, the expression of miR-1297 was low, while the expression of CTGF was elevated. Exosomes derived from BMSCs can inhibit the expression of CTGF by up-regulating the level of miR-1297, thereby inhibiting neuronal cell apoptosis in the hippocampus of depressed rats, while increasing the level of SOD, and reducing inflammatory damage, and ultimately improving the behavioral function of depressed rats.Conclusions:Depressed rats showed decreased expression of miR-1297 and increased expression of CTGF. BMSC-derived exosomes inhibited CTGF expression through up-regulating miR-1297, thereby improving hippocampal neuron damage in rats with depression.

Effect and mechanism of hyperbaric oxygen combined with aquaporin-4 gene silencing on cognitive dysfunction in rats with traumatic brain injury / 中华行为医学与脑科学杂志

Objective:To investigate the effect of hyperbaric oxygen combined with RNA interference (RNAi) technology targeting aquaporin-4 (AQP-4) on improving cognitive function in rats with traumatic brain injury (TBI), and to explore its mechanism.Methods:Totally 112 adult male SD rats were randomly divided into four groups: control group, hyperbaric oxygen(HBO) group, AQP-4 RNAi group and combined treatment group, with 28 rats in each group.The TBI model of rat was established by hydraulic percussion and siRNA targeting aquaporin 4 was constructed. Rats were given corresponding intervention according to their groups.Then the modified neurological severity scores(mNSS)was evaluated on the 7th day and 21th day after operation. Morris water maze test was carried out from the 21st day to 25th day after operation and the percentage of target quadrant and daily escape latency were recorded.The changes of the brain permeability of blood-brain barrier and moisture in brain tissues were measured by Evans blue fluorometry and a wet-dry-weighing technique respectively. The protein expression levels of AQP-4, Caspase-3, Bcl-2, MMP-2 and MMP-9 were detected by Western blot method. The mRNA expression of AQP-4 in TBI brain tissue was measured by RT-PCR method, and the apoptosis rate of TBI brain cells was detected by TUNEL and AnnexinV methods on the 7th day after operation. SPSS 23.0 and Graphpad Prism 7.0 softwares were used for data analysis.One-way ANOVA was used for inter group comparison.Repeated measurement ANOVA was used for Morris results, and the LSD- t test was used for pairwise comparisons. Results:The results of mNSS showed that there were significant differences among the groups on the 7th day and 21st day after operation ( F=4.89, 7.59, both P<0.05). The scores of each treatment group were lower than that of the control group, and the effect of the combined treatment group was the best (7th day: t=3.98, -7.75, both P<0.05; 21st day: t=47.82, 7.94, both P<0.05). The results of Morris water maze test showed that the time and group interaction of rats in the target quadrant residence time and escape latency were not statistically significant( F=1.83, 8.42, both P>0.05). The escape latency and the percentage of stay in the target quadrant in the combined treatment group were better than those in other groups on the 24th and 25th day after operation (all P<0.05). Evans blue staining showed that the contents of Evans blue in AQP-4 RNAi group, hyperbaric oxygen group and combined treatment group were lower than that in the control group(all P<0.05), and that in the combined treatment group was the lowest( t=6.19, P<0.05). The results of dry-wet specific gravity method showed that the water content of brain tissue in the combined treatment group((68.15±1.52)%) was the lowest, and that in the AQP-4 RNAi group((76.71±1.06)%) was lower than that in the HBO group ((80.23±1.43)%)( t=4.38, P<0.05). The results of Western blot showed that the protein levels of AQP-4, Caspase-3, MMP-2 and MMP-9 in the combined treatment group were significantly lower than those in other groups(all P<0.05), while the expression of Bcl-2 was increased in the combined treatment group( P<0.05). RT-PCR results (gray value ratio) showed that AQP-4 mRNA levels in AQP-4 RNAi group(0.61±0.21), HBO group (0.83±0.12), combined treatment group(0.22±0.05) and CON group (1.31 ± 0.25) were significantly different( F=175.05, P<0.05), while the AQP-4 mRNA levels decreased in AQP-4 RNAi group which was better than that in hyperbaric oxygen group ( t=5.25, P<0.05). The decrease was the most obvious in the combined treatment group ( t=58.94, P<0.05). The results of TUNEL and AnnexinV showed that the treatment groups were more effective than the control group in inhibiting neuronal apoptosis, especially in the combined treatment group ( P<0.01). Conclusion:The combination of targeted AQP-4 RNAi and hyperbaric oxgen can effectively promote the recovery of neurological and cognitive function, and the mechanism may be related to protecting the integrity of blood-brain barrier, alleviating brain edema and inhibiting apoptosis of nerve cells after TBI.

Amphetamine causing damage of dopamine cells via inhibiting of protein kinase B / glycogen synthase kinase-3β/ collapsin response mediator protein-2 signal pathway / 解剖学报

Objective To explore the damage mechanism of dopamine cells induced by amphetamine (AMPH). Methods The damage model of dopaminergic cells in mice was established by intraperitoneal injection of AMPH. The mice were randomly grouped into control, saline, amphetamine treatment for 1 day, 7 days, 14 days and 28 days. Each group contained 10 mice. The model of cell injury was established by use of AMPH in PC12 cells. The dopaminergic fibers of corpus striatum and PC12 cells were observed by the immunohistochemistry and immunofluorescence method, and changes of proteins in the protein kinase B (Akt) / glycogen synthase kinase 3β(GSK-3β) / collapsin response mediator protein 2 (CRMP-2) signal pathway were detected by Western blotting. Results AMPH caused the damage of dopaminergic fibers in the mouse corpus striatum and PC12 cells. Meanwhile, AMPH inhibited Akt and GSK-3β phosphorylation levels, and increased phosphorylated CRMP-2 level. Nerve growth factor(NGF), an agonist of Akt, or SB216763, an inhibitor of GSK-3β protected PC12 cells against AMPH-induced toxicity through upregulation of Aat and GSK-3β phosphorylation and downregulated of phosphorylation CRMP-2. Conclusion AMPH causes damage of dopamine cells via inhibition of Akt/ GSK-3β/ CRMP-2 signal pathway.

Regulatory Effect of Liuwei Dihuangtang and Guifu Dihuangtang on Low-dose Hydrocortisone Reduction and Discontinuance in Mice with Drug-induced Deficiency Syndrome / 中国实验方剂学杂志

Objective:To study the effect of the kidney-tonifying recipes on mouse syndrome and adrenal function induced by low-dose hydrocortisone reduction and discontinuation. Method:The 48 male ICR mice were randomly divided into normal control group, hydrocortisone group, Liuwei Dihuangtang group and Guifu Dihuangtang group, with 12 rats in each group. Each group was given 1.32 mg·kg-1·d-1 hydrocortisone for 28 days, and half a dose for 7 days, and the administration was discontinued 14 days before modeling. In the reduction and discontinuation phase, 12.5 g·kg-1·d-1 Liuwei Dihuangtang and 13.5 g·kg-1·d-1 Guifu Dihuangtang were simultaneously administered to the corresponding group for 21 d. On the 28th and 49th day of the experiment, the characteristic information of mice was detected by the experimental methodology of syndrome differentiation and treatment in mice (holding power, body surface infrared temperature). The mice were sacrificed on the 50th day of the experiment, the spleen and thymus were separated, and the spleen and thymus index were calculated, serum corticosterone content is measured by enzyme-linked immunosorbent assay (ELISA), the total RNA of the adrenal gland was extracted, and the expressions of Star, cholesterol-side-chain cleavage enzyme (Cyp11a1), Cyp21a1, Cyp11b1, low-density lipoprotein receptor (Ldlr), scavenger receptor class B type I (Scarb1/SRB1), Hmgcr, acyl-CoA-cholesterol acyltransferase-1 (Acat1), hormone sensitive lipase (Lipe/HSL), insulin-inducible gene 1 (Insig1) and sterol regulatory element binding transcription factor 2 (Srebf2) were detected by Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR). Western blot was used to detect the expressions of LDLR, SRB1, CYP11A1 and cytochrome P450 family 21 subfamily A member 2 (CYP21A2) in adrenal gland. Result:Compared with the normal control group, the body weight and the body mean infrared temperature of the model group were significantly decreased after 28 days of continuous administration (P<0.01). After the administration of the drug, the mice in the model group showed a significant decrease in the grip strength (P<0.05), a significant increase in the thymus index (P<0.01), a significant decrease in the serum corticosterone content (P<0.05). Expressions of adrenal Cyp21a1 and Hmgcr genes decreased (P<0.05, P<0.01), Lipe and Acat1 gene expressions increased significantly (P<0.01), adrenal CYP11A1, SRBI protein expressions decreased significantly, and LDLR protein expression increased. Compared with the model group, the body weight of Guifu Dihuangtang group decreased significantly after 21 days of treatment with traditional Chinese medicine (P<0.01), the spleen index of Liuwei Dihuangtang group decreased significantly (P<0.01), expressions of Cyp11a1, Cyp21a1, Acat1 and Lipe genes in Guifu Dihuangtang group were significantly increased (P<0.05, P<0.01), expressions of Ldlr and Scarb1 genes were significantly decreased (P<0.05), and expressions of Ldlr, Acat1 and Lipe genes in Liuwei Dihuangtang group were significantly decreased (P<0.05, P<0.01), and expressions of CYP11A1 and SRBI proteins in Liuwei Dihuangtang and Guifu Dihuangtang increased. Conclusion:Guifu Dihuangtang can promote the recovery of adrenal function in mice with drug-induced deficiency syndrome by correcting expression of adrenal cortex synthase.

Comparative study on hearing status of preeclampsia infants with different degrees of hypertensive disorder complicating pregnancy and preeclampsia infants without hypertensive disorder complicating pregnancy / 中国实用妇科与产科杂志

OBJECTIVE: To explore the influence and significance of preeclampsia with different degrees of gestational hypertension on infant hearing.METHODS: Totally 1046 newborns whose mothers had a history of pre-eclampsia with hypertensive disorder complicating pregnancy were divided into groups according to the severity of pre-eclampsia and whether they were premature or not from Jan. 2015 to Dec. 2017 in the Forth Hospital of Shijiazhuang.Hearing tests were conducted and compared with newborns whose mothers had no history of pre-eclampsia with hypertensive disorder complicating pregnancy.RESULTS: Of the 14 741 infants studied,57 were finally diagnosed with sensorineural hearing loss,with a total abnormality rate of 0.39%(57/14 741).Among them,34 infants with pre-eclampsia without hypertensive disorder complicating pregnancy had an abnormality rate of 0.25%.There were 12 infants with mild preeclampsia in their mothers,with an abnormality rate of 1.47%.There were 11 infants whose mothers had severe preeclampsia,with an abnormality rate of 4.76%.There were significant differences between mild group and control group,severe group and control group,and severe group and mild group(P0.05),but there was significant difference between the severe group and the control group,and between the severe group and the mild group(P<0.01 and P<0.05,respectively).CONCLUSION: Premature infants with severe hypertensive disorder complicating pregnancy and preeclampsia are at high risk of hearing impairment.The higher the degree of hypertension,the higher the rate of hearing impairment.The emphasis should be put on strengthening the hearing monitoring and follow-up of such children.

Clinical observation of a triple procedure of stimultaneous penetrating keratoplasty combined with extracapsular cataract extraction and intraocular lens implantation / 眼科新进展

Objective To evaluate the operative outcomes of a triple procedure including simultaneous penetration keratoplasty (PKP),extracapsular cataract extraction (ECCE) and intraocular lens (IOL) implantation,and to investigate the relationship between postoperative corneal refractive power and preoperative lens diopter.Methods This retrospective analysis study involved 15 patients who had undergone a triple procedure surgery in Beijing You'an hospital from April to October 2016.Outcomes including the best corrected visual acuity (BCVA),intraocular pressure (IOP),corneal refractive power,axial length,postoperative complications,corneal endothelial cell counts and the survival of corneal graft were determined one year after surgery.Results All corneal grafts were transparent and corneal endothelium were (1974.20 ±472.82) cell · mm-2.The mean postoperative LogMAR visual acuity (0.80 ±0.27) had a significant improvement compared with the mean preoperative LogMAR visual acuity (2.63 ±0.62) (t =13.042,P ＜0.001).There were no statistically significant differences in preoperative IOP (15.27 ± 2.37) mmHg (1 kPa =7.5 mmHg) and postoperative data (14.53 ± 3.04) mmHg (t =0.685,P =0.505),preoperative axial length (23.69 ±2.01) mm and postoperative data (23.62 ±2.12)mm (t =-0.138,P=0.893)and preoperative keratometry (45.01 ± 1.66) D of the control eye and postoperative data (42.56 ± 5.48) D (t =1.202,P =0.260).The postoperative spherical equivalent refractive was (0.40 ±4.65) D,and the target refraction was (0.58 ±0.25)D.Conclusion The triple procedures are safe and effective for the treatment of patients with coexisting corneal pathologies and cataracts.Selection of emmetropia lens diopter may result in the satisfactory postoperative visual acuity.However,unpredictable postoperative corneal curvature changes will still affect the final refractive state.

Research Progress in PTEN as Target Spot in TCM for Prevention and Treatment of Primary Liver Cancer / 中国中医药信息杂志

Phosphatase and tensin homolog deleted on chromosome ten (PTEN) is an important negative regulatory molecule in the development and progression of primary liver cancer and other cancers. TCM has unique advantages in the prevention and treatment of primary liver cancer. This article reviewed the research progress in PTEN as target spot in TCM for prevention and treatment of primary liver cancer in the past 10 years, and provided references for further research and clinical application.

A meta-analysis of effectiveness of influenza vaccine in the elderly in China / 预防医学

Objective To evaluate the protective efficacy of influenza vaccine in the elderly in China. Methods The Chinese databases (CNKI, Wan Fang, VIP) and English databases (Pubmed, Embase) were searched, then studies related to the protective efficacy of influenza vaccine in the elderly according to pre-designed criteria were included and the vaccine efficacy(VE) was selected as an evaluation index. Rev Man 5.3 and Stata 12.0 were used in this meta analysis. Results A total of 26 studies (2000-2016) including 6 kinds of outcomes were eligible, of which, 22 articles related to influenza like illness (ILI) , 5 articles related to common cold (CC) , 11 articles related to the attendance rate due to ILI and CC, 7 articles about chronic diseases (including Hypertension, Diabetes, Coronary Heart Disease (CHD) , Stroke, Cancer, Chronic bronchitis and others) , 6 articles about chronic disease treatment and 3 articles about all-cause mortality. The VE of influenza vaccine was 58.00% (95%CI: 48.00%-66.00%), 40.00%(95% CI: 30.00%-50.00%), 42.00% (95% CI: 34.00%-49.00%), 17.00% (95% CI: 11.00%-23.00%), 28.00%(95% CI: 14 .00 % -40.00 %) and 28 .00 % (95% CI: 15 .00 % -39 .00 %) , respectively. Conclusion Influenza vaccination can effectively prevent the occurrence of influenza like disease and other symptoms in the elderly in China.

Effects of siRNA interference combined with hyperbaric oxygen treatment on cerebral edema and apoptosis in the brain tissue of rats after hemorrhage / 中华老年医学杂志

Objective To examine the effects and mechanisms of siRNA targeting aquaporin 4 (AQP 4) in combination with hyperbaric oxygen therapy(HBO) on cerebral edema and apoptosis in the brain tissue of rats after hemorrhage.Methods Rats were randomly divided into four groups,the control group,the hyperbaric oxygen group,the AQP-4 siRNA group and the combination therapy group (24 rats).Thrombin Ⅶ was injected into the caudate nucleus to establish the hemorrhage model.Construction of siRNA targeting aquaporin 4 was conducted.The mRNA expression of AQP-4 was detected by RT-PCR at day 3.Changes in brain moisture and blood-brain barrier perme ability were measured by a wet/dry weight method and Evans blue fluorometry.The nerve cell apoptosis rate was analyzed by Annexin V andTdT-mediated dUTP-biotin nick end labeling (TUNEL).The expression of proteins including AQP-4,MMP-2,MMP-9,Bcl-2 and caspase-3 was detected by Western Blotting.All the animals were given a score for their nerve function at day 3.Results AQP-4 siRNA treatment obtained better effects than HBO in decreasing the brain edema leveland silencing AQP-4 mRNA(P＜0.05)while,the combination therapy group achieved the best results(P＜ 0.05).Compared with the control group,the percentage of apoptotic cells decreased in all the three treatment groups,with the most marked decrease observed in the combination treatment group(4.24± 0.04)％(F=13.76,P=0.001).The expression of AQP-4,MMP-2,MMP-9 and caspase-3 was lower (P＜0.05) and the expression of Bcl-2 was higher(P＜0.01)in the combination treatment group than in the other three groups.Compared with the control group,all the other three groups received better scores on nerve function defect evaluation at day 3 after hemorrhage(P＜0.05),with the combination treatment group again achieving the most favorable score (4.7 ± 1.1) (F=7.21,P =0.013).Conclusions Targeted siRNA interference combined with hyperbaric oxygen can effectively reduce cerebral edema after cerebral hemorrhage,inhibit neuronal apoptosis and promote neuron function recovery.The underlying mechanisms may be related to down-regulation of AQP-4,MMP 2,MMP-9 and caspase-3 expression and up-regulation of Bcl-2 expression.

Mechanisms of resistance to cancer immunotherapy / 中华微生物学和免疫学杂志

Cancer immunotherapy uses the host′s immune system to mobilize immune cells to rec-ognize and eventually eliminate cancer cells .At present, studies in terms of cancer immunotherapy mainly focus on programmed cell death protein 1/programmed death-ligand 1 (PD-1/PD-L1) antibody, cytotoxic T-lymphocyte-associated protein 4 ( CTLA-4 ) antibody, chimeric antigen receptor T-cell immunotherapy (CAR-T), T cell receptor Immunotherapy (TCR-T), etc.Despite the fact that cancer immunotherapies elicit unprecedented durable responses in clinical therapy , they appear to be ineffective to some patients .In addition, some responders relapse and show resistance to immunotherapies even if their symptoms are re -lieved for a time .Resistance to cancer immunotherapy can be categorized into primary , adaptive and ac-quired, which can occur in every stage during the process of anti-tumor response.In this review, we discuss the known mechanisms of resistance and provide a rationale for the use of combination therapy to overcome resistance.

Effect of Osthole on Adrenocortical Function in Y1 Mouse Adrenocortical Tumor Cells / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the effect of osthole (Ost) on adrenocortical function in Y1 mouse adrenocortical tumor cells.</p><p><b>METHODS</b>Y1 mouse adrenocortical tumor cells were taken as subjects in this experiment. In 10.0%, 1.0%, and 0.1% serum DMEM-F12 medium, Y1 cells were treated with 1, 10, 25, 50, 100, and 200 micromol/L Ost for 24 and 48 h. 0.1% Dimethyl Sulfoxide (DMSO) was taken as negative control group and 1 mmol/L (Bu) 2cAMP as positive control group. Cell growth morphology was observed under inverted microscope. Contents of corticosterone were tested by ELISA. Expression levels of steroids synthase such as Star, Cyp11a1, Cyp21a1, Hsd3b2, Cyp11b1, Cyp11b2, Cyp17a1, and Hsd17b3 mRNA were detected by Real time quantitative PCR (RT-qPCR).</p><p><b>RESULTS</b>Y1 cell proliferation was obviously inhibited by 100 and 200 micromol/L Ost, and its inhibitory effect was more significant in 0.1% serum medium. Compared with the negative control group, gene expressions of Star, Cyp11a1 , Cyp21a1, Hsd3b2, Cyp11b1, Cyp17a1, and Hsd17b3 were significantly enhanced in the posi- tive control group (P < 0.05). Y1 cell corticosterone levels significantly increased in 50 micromol/L Ost treatment group after 24-and 48-h intervention (P < 0.05). Contents of corticosterone increased more obviously in 25 and 50 +/- mol/L Ost treatment groups after 48-h intervention, as compared with 24-h intervention (P < 0.01). After 24-h intervention, expression levels of Star, Cyp21a1, and Hsd3b2 genes were significantly up-regulated in 25 and 50 lLmol/L Ost groups (P < 0.05). Star gene expression was further enhanced after 48-h intervention (P < 0.05). However, Ost showed no effect on Cyp11a1 (P > 0.05). Additionally, gene expressions of Cyp11b1 and Cyp17a1 were significantly enhanced by 10, 25, and 50 pLmolIL Ost after treatment for 24 and 48 h (P < 0.05). Ost showed no obvious effect on Cyp11b2 and Hsd17b3 expressions.</p><p><b>CONCLUSION</b>Ost could regulate adrenal cortex function and promote corticosterone synthesis and secretion through strengthening gene expressions of steroidogenic enzymes.</p>

Inhibition of sciadopitysin against UDP-glucuronosyltransferases / 药学学报

This study was designed to investigate the inhibitory effects of sciadopitysin on the catalytic activities of human 12 kinds of UDP-glucuronosyltransferases (UGTs) in vitro. The risk of drug-drug interactions (DDI) is predicted by in vitro-in vivo extrapolation (IV-IVE). Methods A panel of recombinant human UGT isoforms and human liver microsome (HLM) as well as a series substrates including 4-methyl umbelliferone (4-MU), trifluoperazine (TFP) and N-3-carboxypropyl-4-hydroxy-1, 8-naphthalimide (NCHN) (UGT1A1 specific fluorescent probe substrates) were used to characterize the inhibitory effects of sciadopitysin on human UGTs in vitro. The half maximum inhibitory concentration (IC50) and the constant of inhibition kinetics (Ki) were obtained by nonlinear regression using GraphPad Prism 6.0 software. The potential risk of DDI induced by UGT1A1 was predicted based on in vitro parameters. The results demonstrated that sciadopitysin had strong inhibitory effects on UGT1A1, UGT1A3, UGT1A8 and UGT1A10, with the remaining activity being below 30% at a final concentration of 10 μmol·L-1. For UGT1A1, UGT1A3, UGT1A8 and UGT1A10, the IC50 was 0.20 μmol·L-1 to 1.34 μmol·L-1, the inhibition kinetic constant Ki was 0.07 μmol·L-1 to 2.12 μmol·L-1. The AUC ratio of UGT1A1 can be increased by 19% to 147% at the oral dose of 240 mg·d-1. The sciadopitysin competitively inhibited the formation of 4-MU-O-glucuronide by UGT1A1, UGT1A3, UGT1A8, and UGT1A10. At the same time, the inhibition of NCHN-O-glucuronidation by UGT1A1 was consistent with the competitive inhibition. The strong inhibition of sciadopitysin on UGT1A1 led to reduction of the metabolism of UGT1A1 substrates, and increased the risk of DDI. When co-administrated with other drugs, special attentions should be given to the DDI from inhibition of drug metabolism enzymes to prevent serious clinical consequences.

Establishment of a TMZ-resistant Human Glioma Cell Line U251/TR and the Mechanism of Drug-resistance / 医药导报

Objective To establish a drug-resistance cell line of human glioma with temozolomide ( TMZ) ,investigate its resistance mechanisms, and provide experimental evidence for optimal TMZ therapy. Methods A TMZ-resistant human glioma cell line,U251/TR,was established by stepwise exposure of human parental U251 cells to TMZ. Resistance index and cell viability were accessed by MTT assay. Western-Blot,RT-PCR,immunohistochemistry and immunofluorescence were used to detect MGMT expression for the analysis of resistance mechanism. Results A TMZ-resistant human glioma cell line,U251/TR,was developed after 8 months of stepwise induction with 0. 25-16. 00 μg·mL-1 TMZ. IC50 in U251/TR cells was approximately 7 times higher compared with that in U251 cells (P=0. 00 ). The MGMT expression was significantly increased in U251/TR cells compared with that in parental U251 cells (P=0. 00) . Conclusion A TMZ-resistant human glioma cell line,U251/TR,was established by stepwise exposure of human parental U251 cells to TMZ. The primary mechanism of TMZ resistance is associated with increased activity of MGMT.

Study on pharmacological ingredients of wuzhuyu tang treating migraine by correlating absorption ingredients in everted intestinal sac and pharmacodynamics / 中国中药杂志

Wuzhuyu Tang is a classical formula for treating migraine, but its' pharmacological ingredients is unclear yet. Present study employed the everted intestinal sac model to collect the absorption samples of 10 kinds of Wuzhuyu decoction, and then analyzed the contents of 9 ingredients in Wuzhuyu Tang and absorption samples quantitatively or semi-quantitatively by HPLC-DAD method. Reserpine was used to establish the mice model of migraine, and then the contents and activities of 5-hydroxytryptamine, noradrenaline, dopamine, nitric oxide and nitricoxide synthase in brain tissues and serums were determined respectively after oral administration of Wuzhuyu Tang. Using the partial least squares regression method to correlate the total absorption quantity of 9 ingredients and pharmacodynamics. The result shows that limocitrin-3-O-beta-D-glucoside, ginsenoside Rg1 and Rb1, rutaevine, limonin, evodiamine and rutaecarpine are the main ingredients influenced the effects in absorption samples in everted intestinal sacs, especially ginsenoside Rg1, rutaevine, evodiamine and rutaecarpine among them have obvious improving effects to most pharmacodynamics index, might be the pharmacological ingredients influenced the therapeutical effects of Wuzhuyu Tang treating migraine.

The correlation between adrenal corticosteroids and cytokines expressions in mice of different syndromes / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the cytokines expressions in the adrenal gland and its correlation with serum adrenal corticosteroids in mice of different syndromes.</p><p><b>METHODS</b>Using the quantitative four diagnosis and syndrome differentiation methods, 60 normal mice and 190 H22 liver cancer bearing mice were syndrome typed. Serum corticosterone and aldosterone were tested by ELISA, and mRNA expressions of cytokines in the adrenal gland were detected using Real-time PCR.</p><p><b>RESULTS</b>Mice of different syndromes were obtained, such as normal mice of no syndrome, normal mice of vigorous qi syndrome, normal mice of qi deficiency syndrome, liver cancer bearing mice of excessive evil toxic syndrome, liver cancer bearing mice of evil lying in the middle syndrome, liver cancer bearing mice of weak evil toxic syndrome, and liver cancer bearing mice of poisonous pathogenic factors and qi deficiency syndrome. The serum corticosteroids were significantly higher in the liver cancer bearing mice than in the normal mice (P < 0.05). The cortex hormones increased most significantly in the liver cancer bearing mice of excessive evil toxic syndrome (P < 0.05). Compared with the normal mice, IL-1beta, IL-2, IL-6, IL-10, IL-12alpha, IL-12beta, and TNF-alpha gene expressions increased in the liver cancer bearing mice, while only expressions of IL-1alpha and IL-5 decreased. But the expressions of IL-13 and transforming growth factor beta1 (TGF-beta1) showed no regularity. The expressions of IL-4 and INF-alpha were not detected in all mice. It is notable that the more severe degree of poisonous pathogenic factors, the higher the expressions of serum corticosterone and aldosterone levels as well as IL-6, the lower expressions of IL-1beta, IL-2, IL-5, IL-12alpha, IL-12beta, and TNF-alpha.</p><p><b>CONCLUSIONS</b>The increased serum corticosteroid level in liver cancer bearing mice could possibly be induced by chronic tumor stress, partial cytokines were involved in the synthesis and secretion of the adrenal hormone. Of them, IL-6 might positively regulate the secretion of corticosteroids, while IL-1beta, IL-2, IL-5, IL-12alpha, IL-12beta, and TNF-alpha might negatively regulate their secretions.</p>

Analysis of the dynamic changes of blood hormone levels in H22 liver cancer mice of poisonous pathogenic factors syndromes to different degrees / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the dynamic changes of blood hormone levels in H22 liver cancer mice of poisonous pathogenic factors syndromes (PPFS) to different degrees.</p><p><b>METHODS</b>Two hundred and twenty mice were injected with H22 tumor cells from their armpits. On the ninth day after inoculation the mice of severe poisonous pathogenic factors syndrome (SPPFS) and of mild poisonous pathogenic factors syndrome (MPPFS) were screened. Besides, another normal control group consisting of 30 mice was set up. The mice were killed on the tenth and eleventh day after inoculation (as the 1st and 2nd time window). The weight of the tumor, the wet weight of the thymus and the spleen were weighed. The plasma adrenocorticotropic hormone (ACTH), corticosterone, aldosterone, thyroid hormone T3 and T4, testosterone, TNF-alpha, and IFN-gamma were detected by ELISA. All the aforesaid laboratory parameters were analyzed.</p><p><b>RESULTS</b>The tumor weight was obviously larger in mice of the SPPFS group than in those of the MPPFS group at the same time window (P < or = 0.05). Compared with the normal control group, the thymus was obviously atrophied (P < or = 0.05), the spleen was significantly enlarged (P < or = 0.05), the plasma ACTH significantly increased (P < or = 0.05) in the SPPFS group at the two time windows. But the increment of ACTH was less in the MPPFS group. The plasma corticosterone showed similar tendency as that of ACTH. At the 1st time window the plasma testosterone significantly increased in the two groups (P < or = 0.05). The plasma testosterone and T4 showed a decreasing tendency in the SPPFS group. The plasma TNF-alpha and IFN-gamma levels showed an increasing trend in the two groups. Correlation study showed that the degree of PPFS was negatively correlated with qi deficiency (r = -0.766, P < or = 0.05) and T4 (r = -0.738, P < or = 0.05). The degrees of PPFS was positively correlated with the plasma ACTH level (r = 0.635, P < or = 0.05). The degree of qi deficiency was positively correlated with yang heat syndrome (r = 0.632, P < or = 0.05). The plasma ACTH was negatively correlated with T4 (r = -0.504, P < or = 0.05). The plasma testosterone was positively correlated with TNF-alpha (r = 0.619, P < or = 0.05).</p><p><b>CONCLUSIONS</b>PPFS occurs naturally and shows difference to different degrees in the development of H22 liver cancer. The disorders of neuroendocrine hormones and the suppression of the immune function show dynamic changing trends.</p>

A single institution, retrospective study of treatment experience in primary mediastinal germ cell tumors: elucidating the significance of systemic chemotherapy / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Primary malignant germ cell tumors (GCTs) of mediastinum are rare neoplasms. We introduce our institutional experience in managing patients with primary malignant GCTs of the mediastinum, focusing on the analysis of therapeutic modalities.</p><p><b>METHODS</b>A retrospective review was done in 39 consecutive patients with mediastinal malignant GCTs treated in our institution between 1991 and 2007.</p><p><b>RESULTS</b>A total of 39 patients were enrolled in this study with a median age of 27 years. The 5-year overall survival (OS) and progression-free survival (PFS) rates of the whole population were 60.2% and 57.7%, respectively. Stratified by the histology, 18 patients (46.2%) had seminoma and 21 patients (53.8%) had nonseminomatous germ cell tumors (NSGCTs). The 5-year OS rate of patients with seminoma was 87.4% as compared with 36.7% in patients with NSGCTs (P = 0.0004). The 5-year PFS rate was also significantly higher in seminoma patients (87.4% vs. 31.6%, P = 0.003). For 19 patients with NSGCTs managed with multi-modality treatment, chemotherapy exposure appeared to impact the prognosis. The 5-year OS rate was 44.9% in patients with chemotherapy exposure as compared with 20.0% in patients without it (P = 0.43).</p><p><b>CONCLUSION</b>Our study confirmed the significance of systemic chemotherapy in the treatment of primary mediastinal GCTs.</p>

A method of acutely isolating rat dorsal root ganglion neurons for patch-clamp study of single-channel / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To establish a method of acutely isolating dorsal root ganglion (DRG) neurons for patch clamp study of single-channel.</p><p><b>METHODS</b>DRG neurons of rats were acutely isolated by enzymatic digestion and mechanical blowing.</p><p><b>RESULTS</b>The acutely isolated DRG cells were easy to form the higher sealing resistance (> 5G Omega), which lowered noise level, so that pA-level single channel currents could be recorded.</p><p><b>CONCLUSION</b>The acutely isolated DRG neurons in this study are an ideal for patch-clamp study of single-channel.</p>

Relationship between HBeAg seroconversion with genotypes and HBV specific CTL in patients with chronic hepatitis B treated with Adefovir dipivoxil / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To explore relationship between HBeAg seroconversion with HBV genotypes and HBV specific CTL in patients with chronic hepatitis B (CHB) treated with Adefovir dipivoxil.</p><p><b>METHODS</b>Seventy CHB patients had positive HBV DNA (HBV DNA > or = 1 x 10(4) copy/ml), 45 cases had positive HBeAg, of whom 23 cases (51. 11%) had genotype B, 22 cases (48.89%) had genotype C. ALT > 2 x upper limit of normal value (ULN), human leukocyte antigen (HLA)-A(n) positive, patients were treated with Adefovir dipivoxil (commercial name is Mingzheng, Zhengda Tianjing Pharmaceutical Company), 10 mg, orally, once a day. After treatment for 12 months, observe relationship between HBeAg seroconversion with HBV genotypes and HBV specific CTL.</p><p><b>RESULTS</b>After treatment with Adefovir dipivoxil for 12 months, HBV specific CTL (0.68% +/- 0.11%) was higher than that before treatment (0.33% +/- 0.11%), t = 8.36 P < 0.001, HBV DNA (3.01 +/- 0.2) log10 copy/ml was lower than that before treatment (6.27 +/- 0.70) log10 copy/ml, t = 12.63 P < 0.001, HBV DNA turned negative (< 500 copy/ml) 43 cases (61.43%), in 45 cases with positive HBeAg, HBeAg turned negative in 13 cases (28.89%), 8 cases had HBeAg seroconversion (17.78%), HBV specific CTL (0.86% +/- 0.05%) of patients with HBeAg seroconversion is higher than (0.61% +/- 0.07%) of patients without HBeAg seroconversion (37 cases, 82.22%) t = 7.88, P < 0.001. In 8 cases with HBeAg seroconversion, 7 cases had genotype B (30.43% of genotype B), 1 cases had genotype C (4.55% of genotype C), chi2 = 5.15, P < 0.05.</p><p><b>CONCLUSION</b>Adefovir dipivoxil can enhance HBV specific cellular immunity of CHB patients. After treatment, occurrence of HBeAg seroconversion is related to increase of HBV specific CTL level and may be related to genotypes.</p>