Tangcao pill is commonly applied in adjuvant and even alternative therapy for patients with AIDS. However, the herb contains complex ingredients, but with unknown effect against anti-HIV drug and unknown function. Because CYP450 emzyme is the main metabolic enzymes of the drug, it is of important significance to study the regulation of CYP450 enzymes before and after the combined administration of Tangcao pill and EFV. Proteomics, due to its high throughout and high sensitivity, has been widely applied in CYP450 enzyme study. In this paper, liver microsomes were separated through differential centrifugation. Their proteins were separated through SDS-PAGE. The three protein bands that CYP450 enzymes were located were cut and identified by liquid chromatography tandem mass spectrometry. Totally 16 CYP450 isoenzymes were identified. Furthermore, in order to make a quantitative analysis on the effect of tang herb on CYP450 emzyme, the multiple reaction monitoring (MRM) technology based on MS was adopted. The CYP2C11 was selected based on the results of the mass spectrum identification of proteins. The characteristic polypeptides were obtained through searching Expasy blast database. The m/z of the fragment ions was less than 800. In the paper, the m/z of ion pairs of CYP2C11 were 711.5/232.1, 711.5/319.2, 711.5/466.2 and 711.5/595.3, and the m/z of ESAT-6 (internal standard, IS) were 735.5/215.3, 735.5/389.3, 735.5/460.3 and 735.5/524.3. The relative peak (analyte/IS) area was adopted for the relative quantitative analysis. Compared with the EFV single administration group, the EFV and Tangcao pill combined administration group showed a 1.6-fold increase in CYP2C11. The results of the paper indicated that Tangcao pill may affect drug metabolism by regulating metabolic enzymes such as CYP2C11, but the specific mechanism still unknown.
Animals ; Cytochrome P-450 Enzyme System ; chemistry ; genetics ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Electrophoresis, Polyacrylamide Gel ; Male ; Microsomes, Liver ; chemistry ; drug effects ; enzymology ; Proteomics ; Rats ; Rats, Sprague-Dawley

Objective To know the microcystins pollution caused by cyanobactcria bloom in two drinking water sources in Ningbo city.Methods Water samples were collected at four sampling sites in Yaojiang River and three sampling sites in Miehu reservoir from 20 July,2007 to 31 August,2007.The algae in these water samples were classified and counted,and Microcystin-LR (MC-LR),MC-RR were determined by UPLC-MS-MS.Results The Cyanobacteria cell concentration in Yaojiang River and Meihu reservoir ranged from 0.8?10~6 to 6.7?10~7,and from 1.21?10~7 to 9.928?10~8 cells/L respectively.The MC-LR and MC-RR in Yaojiang River weren't detected,and neither was the MC-RR in MeiHu reservoir.However,the MC-LR was detected at sometimes,and the maximum concentration of MC-LR was 0.84?g/L.Conclusion There is no microcystins pollution in Yaojiang River,but there existed MC-LR pollution in MeiHu Reservoir,and the MC-LR concentration in it was below national standard limit of 1.0?g/L after the treatment of controlling alage and microsystins.

Objective To detect the expression of matrix metallproteinases(MMPs) in aortic valve of children who suffered from rheumatic heart disease(RHD) and to explore the pathological role of MMPs in children′s rheumatic aortic valve disease.Methods RHD group composed of 18 aortic valves from children suffered from RHD.Controls were 8 children who were died accidentally without cardiovascular system diseases.Hematoxylin and eosin stain observing the histological characteristic of the 2 groups.Immunohistochemistry was used to detect expression of MMP2 and MMP9 on aortic valves in 2 groups.Results Hematoxylin and eosin stain showed:in RHD the valves′ structure were destroyed along with fibrous tissue proliferation,mucinous degeneration,collagen and fiber hyalinization,blood vessel and blood capillary proliferation,lymphocyte,plasmocyte,monocyte infiltration.Immunohistochemistry showed that MMP2 and MMP9 expression were significantly higher than those in the aortic of RHD(68.85?13.08,64.35?9.59) compared with control group(107.31?23.39,116.28?6.99)(t=3.92,10.18 all P

OBJECTIVETo analyze the effects of high-frequency electromagnetic field (HF-EMF, 30 MHz, 0-1600 V/m) on the apoptosis and ultramicrostructure of the hippocamp and demonstrate the cytotoxicity of hippocamp.

METHODS120 Wistar female adult rats were randomly divided into ten groups based on body weight with different levels of 30 MHz electromagnetic field (0, 25, 100, 400, 1600 V/m) for eight hours daily. Five group rats were irradiated for three days. The other five group rats were irradiated for fifty-six days. Weekly the rats were continuously exposed five days. The apoptotic rate of the hippocamp was detected with TUNEL System. Meanwhile, the ultramicrostructure was observed with the transmission electron microscope.

RESULTS(1) There was no significant difference on the apoptotic rate and pathological change of the hippocamp cell between the exposure and the control groups through short term experiment (P > 0.05). (2) The apoptotic rate of the granulocyte on the DG campus of the hippocamp in the 400 V/m group and the 1600 V/m group (0.165% +/- 0.049%, 0.189% +/- 0.049% respectively) were increased significantly (P < 0.01) through inferior chronic experiment compared with the control group (0.052% +/- 0.016%). Along with the increase of radiation dose, the ultramicrostructure of the neuron cell appeared more abnormal cells. Especially there were marked change on the neuron in the 1600 V/m group.

CONCLUSIONSThere is no association between cell apoptotic rate of the hippocamp and short period exposure to HF-EMF (30 MHz, 25-1600 V/m). However inferior chronic exposures to HF-EMF might induce the cytotoxicity, especially in the high dose exposure (1600 V/m) under our experiment.

Animals ; Apoptosis ; radiation effects ; Electromagnetic Fields ; Endocytosis ; radiation effects ; Female ; Hippocampus ; cytology ; pathology ; radiation effects ; Neurons ; pathology ; radiation effects ; Rats ; Rats, Wistar

AIMTo evaluate the characteristics, the hypoglycemic efficacy and the pharmacokinetics of the insulin-liposomes double-coated by chitosan (CH) and chitosan-EDTA conjugates (CEC).

METHODSInsulin-liposomes were prepared by reversed-phase evaporation. The protection of insulin against peptic and tryptic digestion was studied with HPLC. The hypoglycemic effects of insulin-liposomes were investigated using the glucose oxidase method after oral administration to rats. Serum insulin concentration in rats were determined by radio-immunoassay, and were assessed by Pkanalyst computer program.

RESULTSThe insulin-liposomes double-coated by CH and CEC was shown to protect insulin against digestion of pepsin, trypsin and gastrointestinal contents. In glucose tolerance test in normal rats, as compared with phosphate buffer solution control group, the insulin-liposomes coated by CH and CEC could reduce the glucose-induced peak of hyperglycemia. The reduction of the insulin-liposomes double-coated by CH and CEC was superior to that of other insulin-liposomes. When administered intragastrically to normal rats, the insulin-liposomes coated by CH and CEC could reduce glycemia measured after an overnight fast. The hypoglycemic effect the insulin-liposomes double-coated by CH and CEC was superior to that of other insulin-liposomes, and the dosage of 50 mu x kg(-1) decreased by 45.98% of initial blood glucose level at 1 h. As compared with subcutaneous injection, the relative pharmacological bioavailability was 17.02% calculated by area under the curve of glucose level versus time profile after oral administration of the insulin-liposomes double-coated by CH and CEC to rats. The serum insulin concentration-time curves were found to best fit the one-compartment open model. As compared with subcutaneous injection, the relative bioavailability was 8.91% calculated by the area under the curve of serum insulin concentration versus time profile after oral administration of the insulin-liposomes double-coated by CH and CEC to rats.

CONCLUSIONThe stability and absorption of insulin-liposomes double-coated by CH and CEC was superior to that of the insulin-liposomes coated either by CH, or by CEC respectively.

Administration, Oral ; Animals ; Biological Availability ; Blood Glucose ; metabolism ; Chitosan ; chemistry ; Delayed-Action Preparations ; Drug Carriers ; Drug Delivery Systems ; Edetic Acid ; chemistry ; Hypoglycemic Agents ; administration & dosage ; pharmacokinetics ; pharmacology ; Insulin ; administration & dosage ; pharmacokinetics ; pharmacology ; Liposomes ; Male ; Nanotechnology ; Particle Size ; Rats ; Rats, Sprague-Dawley ; Technology, Pharmaceutical ; methods

AIMTo evaluate the effect of intracytoplasmic sperm injection (ICSI) in the management of cases with a history of conventional in vitro fertilization (IVF) failure.

METHODSTwo groups of patients, 19 with normal semen parameters and a history of IVF failure (metaphase II oocytes: 0-30 %) and 28 with severe male factor infertility received ICSI technology during the same period. Ovarian stimulation was achieved by conventional procedure. Transvaginal ultrasound-guided oocyte collection was done 35-37 h after human chorionic gonadotrophin (hCG) injection. Only metaphase II oocytes were selected for microinjection.

RESULTSFertilization was achieved with ICSI in all the patients. The fertilization rate (75.6 % +/-21.1 % vs. 73.9 % +/-19.2 %), cleavage rate (85.1 % +/-19.3 % vs. 82.7 % +/-22.1 %), clinical pregnancy rate per embryo transfer cycle (31.6 % vs. 28.6 %) and implantation rate per embryo (15.3 % vs. 14.4 %) did not differ significantly between the two groups.

CONCLUSIONICSI is a valuable method for couples with a history of IVF failure. These patients may have a similar ICSI result as in severe male infertility.

Abortion, Spontaneous ; Adult ; Female ; Fertilization in Vitro ; methods ; Humans ; Infertility, Male ; therapy ; Male ; Oocytes ; Pregnancy ; Pregnancy Rate ; Pregnancy, Ectopic ; Sperm Injections, Intracytoplasmic ; Spermatozoa ; Treatment Failure

Nerve injury produces a long lasting neuropathic pain, manifested as allodynia, a decrease in pain threshold and hyperalgesia, an increase in response to noxious stimuli. The mechanism underlying the lasting abnormal pain is not well understood. Our previous works have shown that electrical tetanic stimulation of the sciatic nerve induces long-term potentiation (LTP) of C-fiber evoked field potentials in the spinal dorsal horn, which is considered as a synaptic model of pathological pain. In the present study we tested if nerve injury, which is proved to produce neuropathic pain, induced the spinal LTP in intact rats. C-fiber evoked field potentials in spinal dorsal horn produced by electrical stimulation (10-20 V, 0.5 ms, 1/min) of the sciatic nerve were recorded. For induction of LTP of C-fiber evoked field potentials, three types of noxious stimuli were applied. (1) Electrical tetanic stimulation (40 V, 0.5 ms pulses at 100 Hz for 1 s repeated four times at 10 s intervals). (2) Transection of the sciatic nerve at 4-5 mm distal to the stimulation electrode. (3) Crushing the sciatic nerve with a forceps four times at 4-5 mm distal to stimulation electrode (from distal to proximal with 1 mm spacing at 10 s intervals), which simulated electrical tetanic stimulation. Acute nerve injury was made by either transection of the sciatic nerve at the distal to the stimulating electrode or crushing the sciatic nerve. We found that nerve injury by cutting or crushing the sciatic nerve produced LTP of C-fiber evoked field potentials lasting until the end of the experiments (3-9 h), and that pretreatment of the sciatic nerve with lidocaine 10 min prior to the nerve transectoin completely blocked LTP induced by nerve transection. The nerve transection-induced LTP was blocked by NMDA receptor antagonist AP5. LTP produced by nerve transection could not be further potentiated by electrical tetanic stimulation, while LTP induced by single electrical tetanic stimulation could be further potentiated by transection of the sciatic nerve. However, when LTP was saturated by several times of electrical tetanic stimulation, nerve transection did not affect the spinal LTP. We conclude that acute nerve injury induces LTP of C-fiber evoked field potentials in intact animals and that nerve transection is more powerful than electrical tetanic stimulation for induction of the spinal LTP. The results further support the notion that LTP of C-fiber evoked field potentials may underlie neuropathic pain.
Animals ; Evoked Potentials ; physiology ; Long-Term Potentiation ; physiology ; Male ; Nerve Fibers, Unmyelinated ; physiology ; Neural Pathways ; drug effects ; physiology ; Nociceptors ; physiology ; Posterior Horn Cells ; enzymology ; physiology ; Rats ; Rats, Sprague-Dawley ; Sciatic Nerve ; injuries ; physiology ; Spinal Cord ; physiology

AIMTo explore the expression of ER and PR mRNAs in endometrium with endometriosis.

METHODSThe rat model of endometriosis was established, and the expression of ER, PR mRNAs in the endometrium was examined by reverse transcription-polymerase chain reaction (RT-PCR).

RESULTSThe expression of ER and PR mRNAs in ectopic endometrium was significantly lower than that in eutopic and normal endometrium (P < 0.01). But no difference was observed between eutopic and normal endometrium (P > 0.05). Ratio of ER/PR mRNA in ectopic endometrium was larger than that in eutopic and in normal endometrium (P < 0.01).

CONCLUSIONThe result illuminates that the increased ER plays a vital role in the onset of endometriosis.

Animals ; Endometriosis ; metabolism ; Endometrium ; metabolism ; Female ; RNA, Messenger ; genetics ; Rats ; Rats, Sprague-Dawley ; Receptors, Estrogen ; genetics ; metabolism ; Receptors, Progesterone ; genetics ; metabolism

AIMTo study the effect of the liposomes coated by chitosan and its derivatives as oral dosage form for peptide drugs on the gastrointestinal (GI) transit of drugs.

METHODSInsulin-liposomes were prepared by reversed-phase evaporation. The in situ perfusion experiment was used to investigate the enteral absorption of insulin. The hypoglycemic effects of insulin were investigated using the glucose oxidase method after administration in rats. The insulin concentrations of serum and enteral tissues were determined by radio-immunoassay in rats.

RESULTSIn in situ local intestinal perfusion experiment, the duodenum was the best segment for the absorption of the insulin liposomes coated by chitosan (CH) or chitosan-EDTA conjugates (CEC) , and double-coated by CH-CEC; the colon was the best segment for the absorption of the insulin solution from rat intestine; but the best segment for the absorption of the uncoated and N-trimethyl chitosan chloride (TMC) coated insulin liposomes was unclear. In all segments, the enteral absorption of the insulin liposomes double-coated by CH-CEC was superior to that of other insulin liposomes.

CONCLUSIONThe insulin-liposomes coated by chitosan and its derivatives can enhance enteral absorption of insulin and increase stability of insulin in GI tract.

Animals ; Area Under Curve ; Chitosan ; chemistry ; Colon ; metabolism ; Delayed-Action Preparations ; Drug Carriers ; Drug Delivery Systems ; Duodenum ; metabolism ; Edetic Acid ; chemistry ; Gastrointestinal Transit ; Hypoglycemic Agents ; administration & dosage ; pharmacokinetics ; Insulin ; administration & dosage ; pharmacokinetics ; Intestinal Absorption ; Liposomes ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo establish an appropriate neonatal rat model of necrotizing enterocolitis (NEC) and to investigate the protective effects of glycomacropeptide (GMP) on the gut from injury in neonatal rats with NEC.

METHODA total of 36 neonatal SD rats were randomly divided into 3 groups: NEC model group (Group M), NEC + GMP group (Group G) and normal control group (Group N), each group had 12 rats. All the neonatal rats were fed with breast milk in the first 3 days after birth. During the second 3 days after birth, the rats of Group N were still maternal breast-fed, but the rats of Group M and Group G were separated from their mothers and lived in incubator and began to be formula fed, and were subjected to cold exposure shortly after hypoxic-reoxygenation treatment. After being fed in such means for 6 days, all the neonatal rats were placed into the incubator and fasted for 24 hours. Then all the rats were sacrificed by cervical dislocation. Intestinal tissue located at the boundary of ileum and cecum was obtained for: (1) histological examination after HE staining, (2) TUNEL detection, (3) electron microscopic observation; and the tissue homogenate was obtained for checking TNF-α and IL-1β levels by ELISA and platelet activating factor (PAF) mRNA expression by quantitative fluorescence (QF)-PCR.

RESULT(1) The pathological scores of the 3 groups were 2.17 ± 0.83 (Group M), 0.92 ± 0.79 (Group G) and 0.17 ± 0.39 (Group N) separately. There was significant difference between Group M and Group G (H = 8.819, P = 0.003). (2) TNF-α levels of 3 groups were (41.94 ± 13.51) pg/ml (Group M), (31.69 ± 11.68) pg/ml (Group G) and (17.42 ± 7.18) pg/ml (Group N) separately, and TNF-α level in Group G was significantly lower than that of Group M (F = 3.959, P = 0.030). (3) IL-1β levels of 3 groups were (150.33 ± 36.41) pg/ml (Group M), (118.36 ± 33.00) pg/ml (Group G) and (28.44 ± 15.04) pg/ml (Group N) separately, and IL-1β level in Group G was lower than that of Group M (F = 5.080, P = 0.013). (4) Expression levels of intestinal PAF mRNA (2(-ΔΔCt) value): 3.01 ± 0.96 (Group M), 1.56 ± 0.29 (Group G), 1.01 ± 0.13 (Group N), the level of Group G was significantly lower than that of Group M (F = 25.251, P = 0.000). (5)Electron microscopy: Group N showed that its cell volume was mostly occupied by the nucleus, the structure was clear, nuclear membrane existed, suggesting the normal phase of cell; Group M showed that apoptotic body existed, suggesting that the advanced stage phase of apoptosis; Group G showed that condensed chromatin marginated around the nuclear envelope, nuclear pores expanded, suggesting the early phase of apoptosis. (6) The apoptosis rate of intestinal epithelial cells by TUNEL detection: 38.79 ± 9.79 (Group M), 29.54 ± 7.30 (Group G), 6.37 ± 1.96 (Group N); the apoptosis rate of intestinal epithelial cells of Group G was significantly lower than that of Group M (F = 6.888, P = 0.003).

CONCLUSIONGMP has protective effects on guts of neonatal rats with NEC, which may probably work by reducing TNF-α, IL-1β and PAF expression, inhibiting the apoptosis of intestinal epithelial cells and reducing intestinal tissue injury.

Animals ; Animals, Newborn ; Apoptosis ; Caseins ; pharmacology ; Cold Temperature ; Enterocolitis, Necrotizing ; drug therapy ; metabolism ; pathology ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Female ; Hypoxia ; complications ; Interleukin-1beta ; metabolism ; Intestinal Mucosa ; metabolism ; pathology ; Intestines ; metabolism ; pathology ; Male ; Peptide Fragments ; pharmacology ; Platelet Activating Factor ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism