Objective To investigate the changes of the expression of GDNF and GDNFR?1 of long-term denervation of posterior cricoarytenoid muscles(PCAMs).Methods 38 patients with vocal paralysis were grouped into four according to their denervated period of time while 12 normal PCAMs as control group.Using double immunofluorescence stain,changes of GDNF and GDNFR1 expression were observed in myofibers at different time points after denervation.Results Double immunofluorescence stain with antibodies against GDNF and GDNFR1 showed no staining in the control group,and study groups.However,after the muscle denervation lasted for 6-12 months and 1-2 years,noted was a significant accumulation of GDNF and GDNFR1 protein in cytolemma and endochylema of myofiber.The mean grey scales and positive region ratios were compared using the image analysis system.The results revealed the levels of GDNF and GDNFR1 protein expression in 6-12 months group,1-2yr group changed significantly(P0.05).Conclusion The changes in expression of GDNF and the acceptor GDNFR-?1,a powerful neurotrophic factor,implied that a good nervous regenerated microenvironment in PCAMs within 2 years.This experiment indicated that denervated posterior cricoarytenoid muscles are able to regain their functions through reinneration within 2 years.

To counteract the immune system in parasitic hosts, some viruses encode proteins to suppress the RNA interference (RNAi) effect. In this report, we established two RNAi systems to be easily observed with strong and obvious effect. The function of the P19 of tomato bushy stunt virus, which suppresses RNAi in mammal cells, was then studied using these two systems. Short hairpin RNAs targeting green fluorescence protein (pshRNA-GFP) and firefly luciferase (pshRNA-luc) were designed and inserted into a eukaryotic transcriptional vector pTZU6+1, respectively. The shRNA expressing vectors were co-transfected with plasmids containing the target gene with or without P19. The GFP expression level was assayed by fluorescence microscopy, Western blotting and RT-PCR. The luciferase expression level was analyzed by the dual-luciferase assay system. pshRNA designed in this study down-regulated the target gene specifically and efficiently, with a decrease of expression of both genes of about 70%, respectively. When P19 was introduced into the RNAi systems, the expression of both GFP and the luciferase were mostly recovered compared with the control groups. The RNAi systems of GFP and luciferase were constructed successfully, demonstrating that P19 of tomato bushy stunt virus has the ability to counteract the RNAi effect induced by shRNA in mammal cells.

In order to determine the suitable harvest time of Dendrobium officinale from different regions in Yunnan province, the drying rate, mannose and glucose peak area ratio, extract, contents of polysaccharide and mannose of D. officinale samples collected from six producing areas in Ynnnan province were determined. The results indicate that drying rate and the contents of polysaccharide and mannose arrived the peak from January to April, extract reached a higher content from September to December, and mannose and glucose peak area ratio from October to February of the coming met the requirment of the Chinese Pharmacopoeia. Hence, the suitable harvesting time of D. officinale in Yunnan province is from December to February of the coming year,according to the experimental results and the request of the Chinese Pharmacopoeia.
China ; Dendrobium ; chemistry ; growth & development ; metabolism ; Glucose ; analysis ; metabolism ; Mannose ; analysis ; metabolism ; Time Factors

To prepare rivastigmine liposome, rivastigmine was loaded into liposome via ammonium sulfate gradient method. Its pharmacokinetic profile in rats was evaluated after intranasal administration. The size, zeta potential, entrapped efficiency and release of rivastigmine from the liposome in vitro were determined. Plasma concentration of rivastigmine was determined by high performance liquid chromatography-tandem mass spectrometry (HPLC/MS) using antipyrine as internal standard. The pharmacokinetic parameters were calculated by DAS 2.0. The entrapped efficiency of rivastigmine liposome was (33.41 +/- 6.58) %, with the mean diameter of 154-236 nm and zeta potential of (-10.47 +/- 2.41) mV. The release behavior of rivastigmine was fitting the first order equation in vitro. The pharmacokinetic studies indicated that the C(max), T(max) and AUC(0-infinity), of rivastigmine liposome were (1.50 +/- 0.15) mg x L(-1), 15 min and (89.06 +/- 8.30) mg x L(-') x min, respectively. Rivastimine liposome was absorbed rapidly, and could reach a certain concentration in rat plasma after intranasal delivery.
Administration, Intranasal ; Animals ; Area Under Curve ; Chromatography, Liquid ; Drug Carriers ; Drug Compounding ; Liposomes ; Male ; Neuroprotective Agents ; administration & dosage ; blood ; chemistry ; pharmacokinetics ; Particle Size ; Phenylcarbamates ; administration & dosage ; blood ; chemistry ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Rivastigmine ; Tandem Mass Spectrometry

Lactoferrin (Lf) is one of the food protein belonged to the innate immune system. Apart from its main biological function of binding and transport of iron ions, lactoferrin also has many other functions and properties such as antibacterial, antiviral, antiparasitic, catalytic, anti-cancer, anti-allergic and radioprotecting. Lf is usually used as additives of food and cosmetics. The research of lactoferrin has been increasingly reported, and the application of lactoferrin as a drug carrier has drawn extensive attention over the recent year. In this paper, researches of lactoferrin as drug carriers are classified and summarized in brain targeting, liver tumor targeting, lung tumor targeting and oral delivery systems according to their different characteristics.
Administration, Oral ; Brain ; Drug Carriers ; Humans ; Lactoferrin ; chemistry ; Neoplasms

OBJECTIVETo compare the differences in the clinical efficacy on Alzheimer's disease between acupuncture and medicine.

METHODSOne hundred and forty-one patients were randomized into an acupuncture group (72 cases) and a medicine group (69 cases). In the acupuncture group, the needling technique for benefiting qi, promoting blood circulation, regulating mind and improving intelligence was used at Shenting (GV 24), Baihui (GV 20), Fengchi (GB 20), Wangu (GB 12), Danzhong (CV 17), Zhangwan (CV 12), Qihai (CV 6), Xuehai (SP 10) and Zusanli (ST 36). The supplementary acupoints were selected according to the symptoms and physical signs. Acupuncture was given once a day and 6 treatments were required for a week. In the medicine group, the choline sterase inhibitor, donepezil (aricept) was prescribed for oral administration, 1 tablet (5 mg) each time, once every night. Four weeks later, the dose was increased to 2 tablets (10 mg) each time. In the two groups, the treatment of 4 weeks made one session and 4 sessions were required. The changes of scores before and after treatment in the minimum mental state examination (MMSE), the activity of daily living scale (ADL), Alzheimer's disease assessment scale-cognition (ADAS-cog) and the digit span (DS) were observed.

RESULTSAfter treatment, scores of MMSE and DS were increased as compared with those before treatment (both P < 0.05) and scores of ADL and ADAS-cog were reduced as compared with those before treatment. The score differences in MMSE, ADL, ADAS-cog and DS before and after treatment were significant in the two groups (all P < 0.01).

CONCLUSIONThe needling technique for benefiting qi, promoting blood circulation, regulating mind and improving intelligence significantly improves the overall function, cognition and activity of daily life in the patients of Alzheimer's disease and the efficacy is better than donepezil.

Activities of Daily Living ; Acupuncture Points ; Acupuncture Therapy ; Aged ; Aged, 80 and over ; Alzheimer Disease ; psychology ; therapy ; Cognition ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome

OBJECTIVETo explore the difference in the efficacy and effect mechanism of subcortical ischemic vascular disease (SIVD) complicated with depression between acupuncture and medication.

METHODSSixty patients were randomized-into an acupuncture group and a medication group, 30 cases in each one. In the acupuncture group, acupuncture was applied to Baihui (GV 20), Shuigou (GV 26), Fengfu (GV 16), Fengchi (GB 20) and the others, once a day, 6 times a week. The treatment of 4 weeks made one session and totally 2 sessions were required. In the medication group, nimodipine 30 mg, three times a day and fluoxetine 20 mg, once a day were prescribed for oral administration, for 8 weeks totally. Before treatment, at the end of the 4th week and at the end of the 8th week of treatment, cerebral blood flow velocity (CBFV) and solubility CD40 ligand (sCD40L) were determined respectively. The scores in Montreal cognitive assessment (MoCA) and Hamilton' s depression scale (HAMD) were evaluated in the two groups. The efficacies on cognitive function and depression symptoms were compared in the patients between the two groups. Results Compared with the outcome before treatment, mean blood flow velocity (Vm) of middle cerebral artery (MCA), anterior cerebral artery (ACA) and posterior cerebral artery (PCA) was increased significantly at the end of the 4th week of treatment in the two groups (all P<0.05). At the end of the 8th week, Vm was increased much significantly (all P<0.01). The differences were not significant in comparison between the two groups (all P>0.05). Compared with the expression before treatment, sCD40L was reduced significantly after treatment in the patients of the two groups (all P<0.01), but the differ- ence was not significant between the two groups (all P>0.05). Compared with that before treatment, MoCA score was increased significantly after treatment in the two groups (all P<0.01), HAMD score was reduced sig- nificantly (all P<0.01), the differences were not significant in comparison between the two groups (all P>0.05). The total effective rate of cognitive improvement was 86.7% (26/30) in the acupuncture group and was 80.0% (24/30) in the medication group, the differences were not significant in comparison of the two groups (P>0.05). The total effective rate of the improvement in depression was 93.3% (21/30) in the acupuncture group and was 86.7% (26/30) in the medication group, the differences were not significant in comparison of the two groups (P>0.05).

CONCLUSIONAcupuncture could significantly increases CBFV and reduces serum sCD40L expressions in the patients of SIVD complicated with depression, and significantly improves cognitive function and relieves depression symptoms. The efficacy of it is similar to that of western medication. The increase of serum sCD40L expression is possibly involved in the occurrence and development of SIVD. Reducing sCD40L expression contributes to the alleviation of damage induced by cerebral ischemia and reperfusion.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Cerebral Infarction ; complications ; drug therapy ; therapy ; Depression ; drug therapy ; etiology ; therapy ; Female ; Fluoxetine ; administration & dosage ; Humans ; Male ; Middle Aged ; Nimodipine ; administration & dosage ; Treatment Outcome

Connexin43 has been shown to play a pivotal role in wound healing process. Wound repair is enhanced by acute downregulation of connexin43, by increasing proliferation and migration of keratinocyte and fibroblast. Angiogenesis is also a central feature of wound repair, but little is known about the effects of connexin43 modulation on functions of endothelial cells. We used connexin43 specific small interference RNA (siRNA) to reduce the expression of connexin43 in human umbilical vein endothelial cell (HUVEC), and investigated the effects of connexin43 downregulation on intercellular communication, viability, proliferation, migration and angiogenic activity of HUVEC. Treatment of siRNA markedly reduced the expression of connexin43 by -80% in HUVEC (P < 0.05), and decreased the intercellular communication by -65% (P < 0.05). The viability, proliferation, migration and angiogenic activity of HUVEC decreased significantly (P < 0.05), compared with that of the normal cells. The results suggest that temporally downregulation of connexin43 expression at early stage of wound to inhibit the abnormal angiogenesis characterized with leaky and inflamed blood vessels, maybe a prerequisite for coordinated normal healing process.
Cell Movement ; Cell Proliferation ; Cell Survival ; Connexin 43 ; metabolism ; Down-Regulation ; Human Umbilical Vein Endothelial Cells ; cytology ; Humans ; Neovascularization, Physiologic ; Umbilical Veins ; cytology ; Wound Healing

The clinical thinking modes of "integrative disease and Syndrome" and "integrative macroscopical Syndrome-differentiation and microscopical Syndrome-differentiation" formed in the past half century were merely the primary train of thought and methodology of integrative traditional Chinese and Western medicine. In order to develop them to a higher level, stress should be put on the unity and holism of disordered organism and unity of function, structure and metabolism. It has long been ignored that a parallel and correlated relationship exists between the TCM Syndrome and pathological process of Western medicine, which is a common, set-formed and regular pathophysiological changes with certain development phase, presented in various diseases. Both have common features as that the same pathological process can be seen in different diseases and several pathological progresses can present in one disease, the same Syndrome also can be seen in different diseases and several Syndromes can present in a disease. Moreover, they have the compatible connotation and denotation. The author deduced that pathophysiological basis of main Syndrome is indeed the basic pathological process. Essence of Syndrome is the related pathological processes including abnormal changes in function, metabolism and morphological structure.
Biomedical Research ; methods ; Clinical Medicine ; methods ; Drug Therapy, Combination ; Holistic Health ; Humans ; Medicine, Chinese Traditional ; methods ; Pathology, Clinical ; methods

Objective To determine if Shenfu injectio(SFI)has any analeptic action in patients emerging from general anesthesia.Methods Eighty-six ASAⅠorⅡadult patients undergoing elective abdominal surgery under general anesthesia were randomly divided into 2 groups(n=43 each):SFI group and control group.The patients were premedicated with intramuscular phenobarbital 0.1g and atropine 0.5mg.Anesthesia was induced with propofol 2 mg?kg~(-1),fentanyl 4-5?g?kg~(-1) and vecuronium 0.1 mg?kg~(-1) and maintained with propofol infusion(2-4 mg?kg~(-1)?h~(-1)),0.5%-1.0% isoflurane inhalation and intermittent i.v.boluses of fentanyl and vecuronium.The patients were intubated and mechanically ventilated.The propofol infusion and isoflurane inhalation were stopped during skin closure.The patients were still unconscious and on mechanical ventilation at the end of surgery and transferred to PACU with a tube in trachea.As soon as the patients reached the PACU,SFI 1 ml?kg~(-1) in Ringer's solution 100 ml was infused over 10 min.In control group the patients received Ringer's solution 100 ml without SFI.The following times were recorded:(1)the time when the patients opened their eyes on command;(2)the time when mechanical ventilation was stopped;(3)the time when oxygen inhalation was stopped;(4)the extubation time;(5)the time of staying in PACU.Venous blood samples were taken before(T_0) and 5,15 and 45 min(T_(1,2,3))after SFI infusion for determination of plasma?-endorphin concentration.Results The awakening time,the mechanical ventilation time,oxygen inhalation time,extubation time and duration of PACU stay were significantly shorter in SFI group than in control group.There were no significant differences in MAP and HR after SFI between the two groups.The plasma?-endorphin concentration was significantly higher in group SFI than in control group.Conclusion Shenfu injectio can make patients emerging from general anesthesia faster.