OBJECTIVE: To investigate the effect of asymmetric dimethylarginine (ADMA), an endogenous inhibitor of nitric oxide synthase, on the activation of hepatic stellate cells (HSCs) and its mechanism. METHODS: Primary HSCs isolated from SD rats were cultured and treated with different concentrations (1, 3 or 10micromol/L) of ADMA for various periods (12 approximately 48h). Expression of alpha-smooth muscle actin (alpha-SMA) and synthesis of type-I collagens in HSC were determined. Messenger RNA levels of the transforming growth factor-beta1 (TGF-beta(1)) in the HSCs were determined using RT-PCR. Intracellular reactive oxidant species (ROS) production was measured using oxidant-sensitive fluorescent indicator. Activation of nuclear factor-kappaB (NF-kappaB) was detected by electrophoretic mobility shift assay (EMSA). RESULTS: ADMA could increase alpha-SMA-positive cells ratio and Type I collagens production of HSCs in a concentration- and time-dependent manner, concomitant with the increase of the TGF-beta(1) mRNA level. Treatment with ADMA (10micromol/L) significantly increased the intracellular ROS production and activated NF-kappaB. Such effects of ADMA on the level of TGF-beta(1) mRNA could be markedly attenuated by pretreatment with antioxidant pyrrolidine dithiocarbamate (25micromol/L). CONCLUSION ADMA can induce the HSC activation by increasing TGF-beta(1) expression through ROS-NF-kappaB-dependent pathway. Therefore, ADMA should be a novel and endogenous activator of HSC, which may be involved in the development of liver fibrosis.
Actins ; biosynthesis ; Animals ; Arginine ; analogs & derivatives ; pharmacology ; Cells, Cultured ; Collagen Type I ; metabolism ; Dose-Response Relationship, Drug ; Gene Expression ; drug effects ; Hepatocytes ; cytology ; drug effects ; metabolism ; Male ; NF-kappa B ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reactive Oxygen Species ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transforming Growth Factor beta ; genetics

Objective: To investigate the effect of rhubarb on gastrointestinal blood perfusion in critical illness and hemorrhagic shocked rats.Methods: Clinical Study: Sixty-four septic patients, who suffered from stress ulcer, were treated with rhubarb at a dose of 25 mg/kg. Twenty-five non-septic patients were taken as control. The gastrointestinal perfusion was evaluated by intramural pH (pHi). Animal study: SD rats were anesthetized with intraperitoneal sodium pentobarbital at a dose of 20 mg/kg. Blood-letting were performed in the animals. Blood pressure reduced to 5.32 kPa and maintained for 120 mins. They were resuscitated at the end of shock by reinfusing all of the shed blood. The rats were randomly divided into four groups: Normal control, shock group, therapeutic group (shocked rats were treated with 50 mg/kg rhubarb at the end of shock) and rhubarb group (normal rats were treated with rhubarb). Laser Doppler was applied to estimate the gastrointestinal blood perfusion. Results: Clinical Study: The gastrointestinal pHi in septic patients was much lower than that in the control, whereas rhubarb could obviously elevate gastrointestinal pHi (P<0.001). In addition, rhubarb also had good effect on gastric hemorrhage caused by stress ulcer. Animal Study: Although the shocked rats were resuscitated completely, their gastrointestinal blood perfusion was much lower than that in the control. Rhubarb could significantly improve the blood perfusion in gastrointestinal mucosa and mesentery (P<0.01). Furthermore, rhubarb also increase the gastrointestinal perfusion in normal rats. Conclusion: Rhubarb could improve gastrointestinal blood perfusion in critical illness and shocked rats.

The complex field cure instrument is a new medical instrument with which an experiment was carried out. Rats were continuously irradiated by the complex field for 90 days, with a day's total dose of 285.9 M.T.G. while other rats weren't irradiated for control group. The animals were respectively killed at 7d, 14d, 30d, 60d and 90d, and their blood samples were taken for cell and humoral immune analysis. The results show that values of lymphocyte transform rate, soluble receptor (SIL-2R), total hemolytic complement levels (CH50) and immunoglobulin (A.G.) after irradiation are more than those of the control group having proved that the instrument may improve immune function of rats.
Animals ; Female ; Immunoglobulin A ; blood ; Immunoglobulin G ; blood ; Immunoglobulins ; blood ; Lymphocyte Activation ; Lymphocyte Count ; Male ; Physical Therapy Modalities ; instrumentation ; Rats ; Rats, Sprague-Dawley ; Receptors, Interleukin-2 ; blood ; Time Factors

BACKGROUNDPreS/S gene was derived from hepatitis B virus (HBV) integration fragment of human hepatocellular carcinoma genome, containing the promoter of preS/S and the C terminal truncated preS/S open reading frame. PreS/S protein may have important roles in processing hepatoma in some HBV-infected patients. The aim of the study was to study the activity of HBV preS/S protein on proliferating cell nuclear antigen (PCNA) promoter and to localize compartment of the preS/S protein in the liver cell line L02.

METHODSThe authors studied the effect of the 3 -truncated preS/S on human PCNA promoter by co-transfecting the expression plasmids of luciferase reporter gene, used the immunohistochemical method to localize the preS/S protein.

RESULTSThe expression product of the plasmid, pKSH7C-HpaI which contained the 3 -truncated preS/S and the flanking cellular sequences, stimulated the expression of PCNA promoter dose dependently,and its effect was 0.5 folds higher than control. Immunohistochemistry showed that the preS/S protein located in the cytosolic region of the liver cell line L02.

CONCLUSIONSThe HBV preS/S protein could stimulate the PCNA promoter of the liver cell, its effect was not direct, which suggests that the effect of preS/S protein on PCNA promoter was probably through the cell signal transduction pathway.

Carcinoma, Hepatocellular ; genetics ; virology ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; genetics ; virology ; Proliferating Cell Nuclear Antigen ; genetics ; Promoter Regions, Genetic ; Protein Precursors ; genetics ; Tumor Cells, Cultured ; Virus Integration

OBJECTIVE: To explore the expression of brain-derived neurotrophic factor (BDNF) in the hippocampus after ovariectomy in mice. METHODS: BDNF levels were detected by immunohistochemistry combined image analysis in hippocampal CA regions and dentate gyrus of ovariectomized mice. RESULTS: The expression of BDNF in hippocampus of mice decreased significantly after the ovariectomy after 4 days. The recovery BDNF expression started 14 days after the ovariectomy and after 28 days, the expression of BDNF in hippocampus recovered to the normal level. CONCLUSION The decrease of estrogen in ovariectomized mice can weaken the expression of BDNF in hippocampus during the early stage.
Animals ; Brain-Derived Neurotrophic Factor ; biosynthesis ; genetics ; Dentate Gyrus ; metabolism ; Estrogens ; blood ; Female ; Hippocampus ; metabolism ; Mice ; Ovariectomy ; RNA, Messenger ; biosynthesis ; genetics

OBJECTIVETo investigate the efficacy of continuous propofol infusion via the common carotid artery for general anesthesia.

METHODSForty adult patients scheduled for abdominal surgery were randomly assigned into 2 groups to receive propopol via the common carotid artery (IC group, n=20) or via the median cubital vein (IV group, n=20). Anesthesia was induced with intravenous administration of drugs and maintained with continuous propofol infusion via the common carotid artery or the median cubital vein, with the CSI stabilized at 40-/+5 till the end of the operation. During the anesthesia, intravenous injection of fentanyl (3 microg.kg(-1).h(-1)) and vecuronium (50 microg.kg(-1).h(-1)) were given intermittently to maintain the analgesia and muscular relaxation. The dose of propofol used, hemodynamics and recovery of the patients were observed.

RESULTSThe dose of propofol used during the surgery to maintain a CSI of 40-/+5 was significantly lower in group IC and than in group IV (2.57-/+0.67 vs 5.72-/+1.37 mg.kg(-1).h(-1), P<0.01). In group IC, the blood pressure was elevated in more than half of the patients and in some cases, the elevation exceeded one third of baseline value and needed intervention with hypotensive drugs. In the IV group, the patients' blood pressure remained stable and varied within the amplitude of 15% of the baseline level. Recovery of spontaneous breathing and consciousness was more quickly in group IC than in group IV (P<0.05).

CONCLUSIONLoss of consciousness and nervous reflex can be achieved with propofol infusion via the common carotid artery, which reduces propofol dose by about 50% in comparison with intravenous infusion and allows more rapid recovery of spontaneous breath and consciousness.

Abdomen ; surgery ; Adult ; Aged ; Analgesics, Opioid ; administration & dosage ; Anesthesia, General ; methods ; Carotid Artery, Common ; Female ; Fentanyl ; administration & dosage ; Humans ; Hypnotics and Sedatives ; administration & dosage ; Infusions, Intra-Arterial ; Male ; Middle Aged ; Nicotinic Antagonists ; administration & dosage ; Propofol ; administration & dosage ; Treatment Outcome ; Vecuronium Bromide ; administration & dosage

OBJECTIVETo investigate the role of mesenchymal stem cells (BMSCs) in multiple myeloma (MM) bone marrow (BM) microenrivonment and their effect on myeloma cells survival and bortezomib induced apoptosis.

METHODSBMSCs were derived from BM of untreated myeloma patients (MM-BMSCs) and healthy donors (HD-BMSCs), respectively. The phenotype, proliferation time and cytokine secretion of MM-BMSCs were detected and compared with HD-BMSCs. Then BMSCs were co-cultured with myeloma cell line NCI-H929 and bortezomib in vitro. The NCI-H929 cells proliferation and bortezomib induced cell apoptosis were investigated.

RESULTSMM-BMSCs and HD-BMSCs were isolated successfully. The phenotype of MM-BMSCs was similar to that of HD-BMSCs. Expressions of CD73, CD105, CD44 and CD29 were positive, but those of CD31, CD34, CD45 and HLA-DR (< 1%) negative. The proliferation time of MM-BMSCs was longer than that of HD-BMSCs (82 h vs 62 h, P < 0.05). Moreover, over-expressions of IL-6 and VEGF in MM-BMSCs culture supernatant were detected as compared with that in HD-BMSCs [(188.8 ± 9.4) pg/ml vs (115.0 ± 15.1) pg/ml and (1497.2 ± 39.7) pg/ml vs (1329.0 ± 21.1) pg/ml, respectively]. MM- BMSCs supported survival of the myeloma cells NCI-H929 and protected them from bortezomib induced cell apoptosis.

CONCLUSIONSMM-BMSCs is benefit for myeloma cells proliferation and against cell apoptosis induced by bortezomib. Over-expression of IL-6 and VEGF maybe play a critical role in these effects.

Apoptosis ; drug effects ; Bone Marrow Cells ; cytology ; Bortezomib ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Multiple Myeloma ; metabolism

OBJECTIVEThe aim of this study was to analyze the complications of coronal incision and investigate the methods of prevention.

METHODSThe retrospective analysis was based on 149 cases, who have had operations since 1997 for congenital craniofacial malformation, second deformation of craniomaxillofacial trauma, maxillofacial tumor or cosmetic purposes.

RESULTSOf them, there were injury of unilateral frontal branch of the facial nerve in 3 cases, subcutaneous hematoma in 9 cases, alopecia in 12 cases, incision scar in 14 cases, obvious strip scar in 2 cases, pains, numbness and paraesthesia in 23 cases, ptosis of facial soft tissue in 8 cases and infection in 4 cases.

CONCLUSIONSThe coronal incision has the merits of distinct exposure, hidden incision scar, but its complications can not be neglected. During the operation, care should be taken to anatomical layers, protecting the blood vessel and nerve bundle in order to reduce complications.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Humans ; Male ; Maxillofacial Injuries ; surgery ; Middle Aged ; Oral Surgical Procedures ; methods ; Postoperative Complications ; prevention & control ; Retrospective Studies ; Scalp ; surgery ; Young Adult

To screen factors related to spermatogonial stem cell (SSC) proliferation, and to investigate the mechanism of infertility caused by cryptorchidism, ten-day-old Kunming (KM) mice were used and experimental cryptorchidism was conducted. On the 35th day after cryptorchid operation, the left testes were fixed in Bouin's fluid and used for histological analysis. The testes of 45-day-old mice were subjected to the same histological analysis, and it was found that they contained germ cells at every stage of development, from SSCs to sperm, indicating that the animals were fully sexually mature at this age. While in experimental cryptorchid mice, the spermatogenesis was arrested at the stage of spermatocytes, and only spermatogonia and primary spermatocytes were present in cryptorchid testes. The proportion of spermatogonia to other types of germ cells was much higher than that in sexually mature mice. On the other hand, the right testes were used for proteomic analysis. The total protein in testes was extracted on the 35th day after cryptorchid operation. The differentially expressed proteins in cryptorchid mice and sexually mature mice were screened and compared by the proteomic techniques. Through the separation of two-dimensional gel electrophoresis (2-DE), 20 differential protein spots were found, and 9 of them were digested and identified by the matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrum. In cryptorchid mice, 6 out of 9 proteins were down-regulated, and 3 were up-regulated. Among these proteins, 4 proteins were identified, and they were Stathmin, phosphatidylethanolamine-binding protein1 (PEBP1), HES-related basic helix-loop-helix protein (HERP), and one unnamed protein (we temporarily named it Px). More Stathmin, PEBP1 and Px were expressed in sexually mature mice than in experimental cryptorchid mice. But HERP1 was the other way round. In the present study, we have screened 4 proteins related to cryptorchidism. It is helpful to study the mechanism of SSC proliferation and infertility caused by cryptorchidism.
Animals ; Cryptorchidism ; metabolism ; Male ; Membrane Proteins ; analysis ; Mice ; Phosphatidylethanolamine Binding Protein ; analysis ; Proteomics ; methods ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; Stathmin ; analysis ; Testis ; chemistry

OBJECTIVETo observe effects of removing arms and ovarian on lumbar intervertebral disc and vertebral bone mineral density (BMD) by establishing rat model of lumbar intervetebral disc degeneration (IDD) with kidney deficiency, and to explore internal mechanism of disc degeneration, relationship between disc degeneration and osteoporosis.

METHODSThirty Sprague-Dawley female rats aged one month were randomly divided into control group, lumbar IDD group and lumbar IDD with kidney deficiency group (combined group), 10 rats in each group. Lumbar IDD group removed double arms, lumbar IDD with kidney deficiency group removed double arms after 3 months, both ovaries were removed. Vertebral bone mineral density were observed by Micro-CT scan; morphological changes were tested by safranine O-fast green staining; II, X collagen protein expression in the intervertebral disc were obsevered by immunohistochemistry; extracellular matrix gene expression were obsevered by real-time polymerase chain reaction (RT-PCR), in order to evaluate the effects of removed of forelimbs and double ovarian on degeneration and vertebral bone mineral density of intervertebral disc.

RESULTSMicro-CT scan showed osteoporosis in kidney deficiency group was obviously worse than other two groups; safranine O-fast green staining showed that intervertebral space became narrowed, intervertebral disc tissue degenerated obviously, chondral palte was underdeveloped in kidney deficiency group; immunohistochemistry showed that X collagen expression increased, type II collagen expression decreased in kidney deficiency group; RT-PCR showed that type II collagen expression in lumbar IDD group and kidney deficiency group was lower than control group, and had statistical meaning among three groups (P=0.000, P=0.000); Age 1 in lumbar IDD group and kidney deficiency group was lower than control group, and had statistical meaning among three groups (P=0.000, P= 0.000); while type X collagen expression was higher than control group, but no significant meaning; MMP-13 in lumbar IDD group and kidney deficiency group was higher than control group, with significant meaning compared among three groups (P= 0.000, P=0.000); aggrecanase-2 in lumbar IDD group and kidney deficiency group was higher than control group, with significant meaning compared among three groups (P=0.006, P=0.008).

CONCLUSIONRats model of lumbar disc degeneration established by removed forelimbs and ovariectomized can occure "bone like"--osteoporosis, which is similar with clinical kidney lumbar disc degeneration in tissue morphology, molecular cell biology expression.

Animals ; Collagen ; genetics ; metabolism ; Extracellular Matrix ; genetics ; metabolism ; Female ; Humans ; Intervertebral Disc Degeneration ; etiology ; metabolism ; physiopathology ; surgery ; Kidney ; physiopathology ; Osteoporosis ; complications ; genetics ; metabolism ; Ovariectomy ; adverse effects ; Rats ; Rats, Sprague-Dawley