Objective To explore the values of translocator protein (TSPO) and serum cell-free mitochondrial DNA (cf-mtDNA) in predicting the disease condition and prognosis of patients with traumatic shock. Methods Eighty patients (traumatic shock group) with traumatic shock and eighty patients (without traumatic shock group) without traumatic shock were selected. Complete demographic and clinical laboratory data of patients were collected. Blood samples of patients with traumatic shock were collected at the time points of immediately after admission (T₁) and the first day (T₂), the third day (T₃) and seventh day (T₄) after admission, the level of TSPO was measured by enzyme-linked immunosorbent assay (ELISA), and level of cf-mtDNA was measured by quantitative real-time polymerase chain reaction (qPCR). The levels of TSPO and cf-mtDNA were compared between patients with and without traumatic shock; the patients with traumatic shock were divided into the poor prognosis group and good prognosis group according to differed prognostic outcome, and the levels of TSPO and cf-mtDNA were compared between the two groups. The predictive values of TSPO and cf-mtDNA for the prognosis of patients with traumatic shock were analyzed by the receiver operating characteristic (ROC) curve. Results Compared with the no traumatic shock group, the traumatic shock group had higher levels of serum TSPO at T₁ to T₄ and higher levels of cf-mtDNA at T₂ to T₃, and the differences were statistically significant (P < 0.05). Compared with the good prognosis group, the poor prognosis group had higher levels of serum TSPO and cf-mtDNA at T₁ to T₄, and the differences were statistically significant (P < 0.05). Taking the prognosis of traumatic shock patients as the state variable to perform ROC curve analysis, the results showed that the area under the curve (AUC) of TSPO from T₁ to T₄ for predicting prognosis in the poor prognosis group was 0.825, 0.829, 0.695 and 0.869 respectively, while those of cf-mtDNA level from T₁ to T₄ for predicting prognosis in the poor prognosis group was 0.766, 0.766, 0.837 and 0.783 respectively, and the differences were statistically significant (P < 0.001). Conclusion Traumatic shock patients have elevated levels of TSPO and cf-mtDNA, with significantly higher levels observed in those with poor prognosis, and the TSPO level on the seventh day and cf-mtDNA level on the third day have the highest predictive value for prognosis.

China ; Humans ; Lenalidomide/therapeutic use* ; Multiple Myeloma/drug therapy* ; Thalidomide ; Therapeutic Equivalency

Objective To observe and compare the clinical efficacy of fire needling versus electroacupuncture in treating knee osteoarthritis of kidney and marrow deficiency type.Methods One hundred and ten patients with knee osteoarthritis of kidney and marrow deficiency type were randomly allocated to two groups, a fire needling group of 56 cases and an electroacupuncture group of 54 cases. The fire needling group received filiform needle acupuncture and fire needle pricking and the electroacupuncture group, electroacupuncture. Treatment was given once every other day, for a total of four weeks as one course. The Visual Analogue Scale (VAS) score and the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score were recorded in the two groups before and after treatment. The clinical therapeutic effects were compared between the two groups.Results The VAS score and the WOMAC subscores significantly decreased in both groups after treatment compared with those before treatment (P<0.01). There were no statistically significant post-treatment differences in the VAS score and the WOMAC subscores between the two groups (P>0.05). The VAS score and the WOMAC subscores decreased further in both groups at four weeks after treatment compared with those at the end of treatment (P<0.01). In the electroacupuncture group, there were statistically significant differences in the VAS score and the WOMAC pain and stiffness subscores between eight weeks and four weeks after treatment (P<0.01) and in the WOMAC pain, stiffness and joint function subscores between at eight weeks after treatment and at the end of treatment (P<0.01). In the fire needling group, there were statistically significant differences in the VAS score and the WOMAC pain, stiffness and joint function subscores between at eight weeks after treatment and at the end of treatment (P<0.01). At four weeks after treatment, the VAS score and the WOMAC pain, stiffness and joint function subscores were significantly lower in the fire needling group than in the electroacupuncture group (P<0.05,P<0.01). At eight weeks after treatment, the VAS score and the WOMAC pain and stiffness subscores were still significantly lower in the fire needling group than in the electroacupuncture group (P<0.01). The total efficacy rate was 94.6% in the fire needling group and 90.7% in the electroacupuncture group; there was no statistically significant difference between the two groups (P>0.05).Conclusions Both filiform needle acupuncture plus fire needle pricking and electroacupuncture can markedly relieve the pain and stiffness and improve joint function in knee osteoarthritis of kidney and marrow deficiency type. Both have an equivalent short-term therapeutic effect. Filiform needle acupuncture plus fire needle pricking has a better long-term therapeutic effect than electroacupuncture.

BACKGROUND/AIMS: This study was to investigate whether transforming growth factor-β1 (TGF-β1) plays a role in hyperalgesia in chronic pancreatitis (CP) and the underlying mechanisms. METHODS: CP was induced in male adult rats by intraductal injection of trinitrobenzene sulfonic acid (TNBS). Abdominal hyperalgesia was assessed by referred somatic behaviors to mechanical stimulation of rat abdomen. Dil dye injected into the pancreas was used to label pancreas-specific dorsal root ganglion (DRG) neurons. Whole cell patch clamp recordings and calcium imaging were performed to examine the effect of TGF-β1 on acutely isolated pancreas-specific DRG neurons. Western blot analysis was carried out to measure the expression of TGF-β1 and its receptors. RESULTS: TNBS injection significantly upregulated expression of TGF-β1 in the pancreas and DRGs, and TGF-β1 receptors in DRGs (T9-T13) in CP rats. Intrathecal injection of TGF-β receptor I antagonist SB431542 attenuated abdominal hyperalgesia in CP rats. TGF-β1 application depolarized the membrane potential and caused firing activity of DRG neurons. TGF-β1 application also reduced rheobase, hyperpolarized action potential threshold, and increased numbers of action potentials evoked by current injection of pancreas-specific DRG neurons. TGF-β1 application also increased the concentration of intracellular calcium of DRG neurons, which was inhibited by SB431542. Furthermore, intrathecal injection of TGF-β1 produced abdominal hyperalgesia in healthy rats. CONCLUSIONS: These results suggest that TGF-β1 enhances neuronal excitability and increases the concentration of intracellular calcium. TGF-β1 and its receptors are involved in abdominal hyperalgesia in CP. This and future study might identify a potentially novel target for the treatment of abdominal pain in CP.
Abdomen ; Abdominal Pain ; Action Potentials ; Adult ; Animals ; Blotting, Western ; Calcium ; Diagnosis-Related Groups ; Fires ; Ganglia, Spinal ; Humans ; Hyperalgesia ; Injections, Spinal ; Male ; Membrane Potentials ; Neurons* ; Pancreas ; Pancreatitis, Chronic* ; Rats*

Objective: To obtain purified and functional CDNF-his recombinant protein and prepare its polyclonal antibodies.Methods:Preparation of recombinant CDNF-his was carried out in HEK 293 T cells with pVR1012-CDNF-his successfully constructed transfected into them.Then,the recombinant protein was purified by Ni-NTA immunoaffinity chromatography.The purity was analyzed by SDS-PAGE and the protein′s identity was tested by Western blot.MTT was used to verify the biological function of the protein purified.New Zealand white rabbits were immunized with purified CDNF-his protein for preparation of polyclonal antibodies.Results:pVR1012-CDNF-his expressed successfully in HEK 293 T cells.The purity of protein was up to more than 90%after purification.MTT showed that CDNF-his was able to protect PC 12 cells from damage by 6-OHDA.The polyclonal antibody was detected at the end of animal immunizing process.Conclusion: A method to express and purify protein using HEK 293T cell and following Ni-NTA immunoaffinity chromatography has been built.CDNF-his with biological activity is obtained based that.Finally, polyclonal antibodies of CDNF were generated successfully.

The UL49.5 gene of most herpesviruses is conserved and encodes glycoprotein N. However, the UL49.5 protein of duck enteritis virus (DEV) (pUL49.5) has not been reported. In the current study, the DEV pUL49.5 gene was first subjected to molecular characterization. To verify the predicted intracellular localization of gene expression, the recombinant plasmid pEGFP-C1/pUL49.5 was constructed and used to transfect duck embryo fibroblasts. Next, the recombinant plasmid pDsRed1-N1/glycoprotein M (gM) was produced and used for co-transfection with the pEGFP-C1/pUL49.5 plasmid to determine whether DEV pUL49.5 and gM (a conserved protein in herpesviruses) colocalize. DEV pUL49.5 was thought to be an envelope glycoprotein with a signal peptide and two transmembrane domains. This protein was also predicted to localize in the cytoplasm and endoplasmic reticulum with a probability of 66.7%. Images taken by a fluorescence microscope at different time points revealed that the DEV pUL49.5 and gM proteins were both expressed in the cytoplasm. Overlap of the two different fluorescence signals appeared 12 h after transfection and continued to persist until the end of the experiment. These data indicate a possible interaction between DEV pUL49.5 and gM.
Animals ; Ducks/virology ; Genes, Viral/genetics ; Mardivirus/*genetics ; Membrane Glycoproteins/*genetics ; Microscopy, Fluorescence ; Phylogeny ; Polymerase Chain Reaction/veterinary ; Viral Envelope Proteins/*genetics

Objective To explore the aesthetic efficacy of modified endoscopic rhytidectomy by using the techniques to minimize tissue damage,to obviate injury to the vessels and nerves,and to control bleeding and to firm fixation.Methods Two discontinuous incisions were made in the temporal scalp during the procedure,obviating injury to the branches of the superficial temporal vessels.Endoscopic technique was used to facilitate elevating,hemostasis,slinging and fixation in the plane under superficial temporal fascia.Three transverse incisions were made after the hairline in the forhead scalp,the operation was carried out by using endoscopic equipment,and the elevated forhead flap was slinged and fixed upward to the lamina externa cranii.Results 58 cases were received endoscopic forehead and temporal rhytidectomy,only slight edema was observed after surgery,and no obvisous ecchymosis was found.All patients returned home 7 days after operation.Degree of satisfaction on long-term follow-up showed that 56 cases(96.55%)improved obviously one year postoperatively;35 cases followed up 2 years,33(94.29%)of them improved obviously.None case was suffered from facial nerve injury.Conclusion The purpose of endoscopic rhytidectomy is to avoid carrying out the operation out of sight,to minimize unexpected damage to vessels and nerves,and to facilitate dissection,hemostasis,suturing,slingling and fixation.By refining the technique,we can achieve minimal injury,shorten recovery period,and obtain more satisfactory results,so the indication for operation is extended.

The differentiation of neural stem cells/neural precursors (NSCs/NPs) is a hot spot in neurobiological research. It used to identify their differentiation degree only by morphologic appearances. The functional characteristics, such as electrical properties of cellular membrane and ion channel activities, are drawing more and more attention with the development of patch clamp technique. It was summarized the recent progress in the study of NSCs/NPs functional differentiation using patch clamp, some existing problems and research perspectives were suggested.

BACKGROUND: Changes of elastic fibers in middle cerebral artery(MCA)is close related withthe aged cerebrovascular disease.OBJECTIVE: To observe the changes of elastic fibers of MCA in different aging rats.DESIGN: A descriptive and controlled study based on experimental animals.SETTING: Department of anatomy and central laboratory in a university.MATERIALS: Totally 36 healthy Wistar rats with either gender, weighing 200 - 280 g, were selected from the Animal Institute of the third medical military university of Chongqing[certification SCXX (army) 2002-007].INTERVENTIONS: Changes of elastic fibers of MCA of different aging rats were observed with light microscope, transmission electron microscope and image analysis system. MAIN OUTCOME MEASURES: ①) Major outcome: changes of elastic lamella in MCA of different aging rats; ②) Secondary outcome: ultramicrostructural changes of internal lamella under the transmission electron microscope. RESULTS: With the increase of age, the folded extent and quantity of internal elastic lamella were decreased, and the content of elastic fibers were also decreased significantly( P ＜ 0.01 ). However, the ratio of collagen fibers to elastic fibers was increased significantly( P ＜ 0.01 ) . In the aging group above 24 months, the internal elastic lamina thinned, delaminated and disrupted, and the lipid deposited in it. Endothelial cells and smooth muscle cells passed through the internal elastic lamina. CONCLUSION: Changes of elastic fibers may be related with the increased susceptibility to the cerebrovascular disease in aged people.

This study was conducted to examine the effectiveness of a gene transfer of human TGF beta 1 gene into smooth muscle cells and whether the TGF beta 1 can increase elastin expression of smooth muscle cells. With the help of DOTAP, smooth muscle cells were transfected with pMAMneoTGF beta 1. The positive cell clones were selected with G418. The stable transfection and expression of TGF beta 1 in the smooth muscle cells were determined by immunofluorescence analysis. The expression of elastin in the transfected and untransfected cells were determined by in situ hybridization. The adhesion force between smooth muscle cells and matrix was detected by micropipette system. The results showed abundant TGF beta 1 stable expression in smooth muscle cells. TGF beta 1 gene can increase two-three times elastin expression and increase the adhesion between smooth muscle cells and matrix. TGF beta 1 can be used in vascular tissue engineering to increase smooth muscle cells adhesion.
Cell Adhesion ; Cells, Cultured ; Elastin ; biosynthesis ; Humans ; In Situ Hybridization ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Transfection ; Transforming Growth Factor beta ; biosynthesis ; genetics ; physiology ; Transforming Growth Factor beta1