Objective: To explore the molecular mechanism of the supplemented Naotaifang (sNTF) in the treatment of vascular dementia (VD), and the mRNA expression profiles of hippocampal tissue of VD model rats before and after the intervention of modified sNTF were investigated by microarray analysis. Methods: VD model was established by bilateral common carotid artery ligation. VD rats were treated with sNTF for 30 days. HE staining and Morris water maze were used to evaluate the therapeutic effect of sNTF. The mRNA expression profiles data of VD model rats before and after intervention of sNTF were obtained by Agilent mRNA expression chip. The significantly differentially expressed genes were screened by microarray analysis, and the protein-protein interaction (PPI) network was constructed. The biological processes and signaling pathways in which differentially expressed genes were mainly involved and analyzed by GO and pathway enrichment. Immunohistochemistry and qRT-PCR were used to verify the chip analysis results. Results: HE staining and Morris water maze experiments showed that VD rats showed cerebral ischemia, hippocampal neuron damage, and decreased spatial learning and memory function, but sNTF can partially reverse this trend. 469 differential expression genes were screened by microarray analysis, including 180 up-regulated genes and 289 down-regulated genes. IL6, FGF2, TNF, and IL1b may be the main pharmacodynamic targets of sNTF in the treatment of VD rats, and the results were verified by immunohistochemistry and qRT-PCR. GO and pathway enrichment analysis showed that these genes were closely related to biological processes such as inflammation and apoptosis, and these genes were mainly involved in the regulation of TNF signaling pathway, toll-like receptor signaling pathway and the apoptosis pathway. Conclusion: The results suggested that the therapeutic effect of DSS on AD involves multiple genes and pathways, and and inhibition of hippocampal neuroinflammation may be one of the important mechanisms of its anti-VD.

OBJECTIVETo observe the therapeutic angiogenesis effect of naotai recipe (NR) on local ischemia/reperfusion (I/R) injury of rats by observing signaling pathway of hypoxia-inducible factor-lα (HIF-1α) and vascular endothelial growth factor (VEGF).

METHODSTotally 120 Sprague-Dawley (SD) rats were randomly divided into 4 groups, namely, the normal control group (n =12), the sham-operation group (n =12), the I/R model group (n =48), and the NR group (n =48). Cerebral I/R injury models were established using thread suture method. Rats in the I/R model group and the NR group were sub-divided into 4 sub-groups according to the 1st, 3rd, 5th, and 7th I/R day (n =12). The phenomenon of neovasculization was observed by immunofluorescence staining. The protein and mRNA expression levels of HIF-la, VEGF-A, and VEGFR II receptor were detected by RT-PCR.

RESULTSThere were a large amount of labels for neovasculization in the ischemic area of the NR group. Double-immunofluorescence labeling [vWF (red) and BrdU (green)] was observed in the NR group. Compared with the model group, the HIF-1α protein expression was obviously enhanced on the 1 st day of I/R (P <0.01), and the VEGF protein expression started to enhance on the 3rd day in the NR group (P <0.01). The VEGFR protein expression level was the highest in the NR group on the 5th day of I/R (P <0.01). The protein expression of VEGF and HIF-1α started to decrease on the 7th day of I/R.

CONCLUSIONNR could strengthen angiogenesis after I/R by elevating the expression of HIF-lα and activating HIF-lα/VEGF signaling pathway.

Animals ; Brain Ischemia ; metabolism ; Cerebral Infarction ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Hypoxia-Ischemia, Brain ; metabolism ; Ischemia ; Neovascularization, Pathologic ; Rats, Sprague-Dawley ; Reperfusion Injury ; Signal Transduction ; Vascular Endothelial Growth Factor A ; biosynthesis

OBJECTIVETo assessment result with multi-measures operation for adhering otitis media.

METHODSA retrospective review of 108 cases (male 45, female 63), double ears 12, all was 120 ears. Of 20 ears had been radical operated for cholesteatoma some years ago. Of 46 ears had incorporate cholesteatoma now. Of 54 ears had only adhering in middle ear but no had been radical or incorporate cholesteatoma who underwent operation from January 1999 to May 2003 for chronic ear disease as well as ossiculoplasty with multi-technique measures (for example auto-bone cup-pole or gung-pole as total and partial ossicular replacement prostheses; scute rebuild by auto-bone; absorbable Metrogel; tympanic chorda nerve as a spring press prostheses etc) were used in operation, and were followed up mean (26.11 +/- 11.31) months (range 12 approximately 53 months).

RESULTSOne hundred and twenty ears mean AB gap was (38.60 +/- 12.70) dB pre-operation. It was (15.28 +/- 11.69) dB post-operation. Partnership t test, t = 21.24, t 0.01 = 2.60 < 21.24, P < 0.01. The air-bone gap (AB gap) closure to within 20 dB as succeed achieved in 70% (84 ears). AB gap closure in 10 dB as best result in 40% (48 ears). Air conduction was under 40 dB in 70% (83 ears). AB gap closure not enough 10 dB and Air conduction over 40 dB as no effect was 6% (7 ears) .

CONCLUSIONSAs long as technique measures is right, even though adhering middle ear had much difficult in operation, it still can gain succeed to 70%. But had still 30% no succeed, more study is necessary.

Adolescent ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Otitis Media with Effusion ; pathology ; surgery ; Retrospective Studies ; Tissue Adhesions ; pathology ; surgery ; Treatment Outcome ; Tympanoplasty ; Young Adult

OBJECTIVETo observe different effects of moxibustion on extracellular potassium ion in acupoint under physiological and pathological status and provide experimental evidence for exploring action mechanism of moxibustion on acupoint local.

METHODSForty female SD rats were randomly divided into a blank group, a blank-moxibustion group, a model group and a model-moxibustion group, 10 cases in each one. The complete Freund's adjuvant(CFA) was adopted to establish model of adjuvant arthritis (AA) in the model group and model-moxibustion group. No treatment was given in the blank group and model group while moxibustion was applied at "Zusan-li" (ST 36) for 30 min in the blank-moxibustion group and model-moxibustion group. The tissue fluid in "Zusanli" (ST 36) was collected with microdialysis and real-time analyzed by electrolytic analyzer. The change of concentration of potassium ion in "Zusanli" (ST 36) was observed.

RESULTS(1) Under physiological status, the concentration of extracellular potassium ion in the blank group was not changed within 150 min (P > 0.05); before the moxibustion, the concentration of extracellular potassium ion in the blank-moxibustion group was (1.21 +/- 0.31) mmol/L, and after treatment it was gradually increased and reached its peak at (2.38 +/- 0.42) mmol/L after 60 min (P < 0.05), then it was reduced. 150 min after the treatment, concentration of potassium ion was slightly higher than that before moxibustion as well as that in the blank group. The concentration in the blank-moxibustion group at 60 min was statistically significant compared with that in the blank group (P < 0.05). (2) Under pathological status, the concentration of extracellular potassium ion in the model group was not changed within 150 min, differences of which at each time point was not statistically significant (all P > 0.05). Before the moxibustion, the concentration of extracellular potassium ion was (1.09 +/- 0.12) mmol/L in the model-moxibustion group, and it was immediately increased to (1.96 +/- 0.18) mmol/L after moxibustion. 60 min and 90 min after the moxibustion, it still maintained a higher level, which was (1.87 +/- 0.29) mmol/L and (1.59 +/- 0.16) mmol/L respectively (both P < 0.05). The differences of each time point after moxibustion in the model-moxibustion group were statistically significant compared with those in the model group (all P < 0.05).

CONCLUSIONThe moxibustion could increase the concentration of potassium ion in rat's acupoint local under physiological status but time of effect is short; with moxibustion at "Zusanli" (ST 36) under pathological status, the concentration of local potassium ion is obviously increased and maintains for a long time.

Acupuncture Points ; Animals ; Arthritis, Experimental ; metabolism ; therapy ; Disease Models, Animal ; Female ; Humans ; Moxibustion ; Potassium ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To investigate the expression and clinical significance of matrix met-alloproteinases-9 (MMP-9)in tissues of endometrial carcinoma.Methods The paraffin samples of tissues in 30 patients with endometrial carcinoma admitted in our hospital from February 2009 to March 2012 were selected as experimental group 1 while those in other 40 patients with atypical en-dometrial hyperplasia and 40 healthy people as experimental group 2 and control group,respective-ly.Immunohistochemical staining method was used for the MMP-9 staining so as to analyze the counts of cells with positive staining and the staining severity.MMP-9 expression levels were com-pared,the relationship between MMP-9 expression and clinical pathological parameters in endome-trial carcinoma tissues were analyzed and Cox ratio risk model was applied to analyze the prognostic factors of endometrial carcinoma.Results Experimental group 1 was evidently lower in low-expres-sion of MMP-9 than experimental group 2 and apparently higher in high-expression of MMP-9 than the other groups (P <0.05).It was showed that pathological staging and lumph node metastasis were closely associated with MMP-9 protein expression (P <0.05),which were also the risk fac-tors impacting the prognosis of endometrial carcinoma (P <0.05).Conclusion The specific high-expression of MMP-9 in endometrial carcinoma can be considered as one of the referent indexes for diagnosing endometrial carcinoma and its pathological staging,which,however,is not the indepen-dent risk factor influencing the prognosis of endometrial carcinoma.

Objective To investigate the expression and clinical significance of matrix met-alloproteinases-9 (MMP-9)in tissues of endometrial carcinoma.Methods The paraffin samples of tissues in 30 patients with endometrial carcinoma admitted in our hospital from February 2009 to March 2012 were selected as experimental group 1 while those in other 40 patients with atypical en-dometrial hyperplasia and 40 healthy people as experimental group 2 and control group,respective-ly.Immunohistochemical staining method was used for the MMP-9 staining so as to analyze the counts of cells with positive staining and the staining severity.MMP-9 expression levels were com-pared,the relationship between MMP-9 expression and clinical pathological parameters in endome-trial carcinoma tissues were analyzed and Cox ratio risk model was applied to analyze the prognostic factors of endometrial carcinoma.Results Experimental group 1 was evidently lower in low-expres-sion of MMP-9 than experimental group 2 and apparently higher in high-expression of MMP-9 than the other groups (P <0.05).It was showed that pathological staging and lumph node metastasis were closely associated with MMP-9 protein expression (P <0.05),which were also the risk fac-tors impacting the prognosis of endometrial carcinoma (P <0.05).Conclusion The specific high-expression of MMP-9 in endometrial carcinoma can be considered as one of the referent indexes for diagnosing endometrial carcinoma and its pathological staging,which,however,is not the indepen-dent risk factor influencing the prognosis of endometrial carcinoma.

Porcine epidemic diarrhea virus (PEDV) LJB/03 strain was isolated from the feces of piglets suspected to be suffering from a severe diarrhea in Heilongjiang Province, and was identified by immunofluorescence test, immunelectronmicroscopy, RT-PCR and indirect ELISA assay. Characteristics of the virus culture and the methods of improvement of virus titer were explored. The results showed that the virus had the typical appearance of the coronavirus. Analysis of the nucleotide sequences of RT-PCR products revealed 98% homology with the reference strains. Indirect immunofluorescence assay showed a significant presence of green fluorescence, and an average P/N ratio of 7.6 by indirect ELISA assay. Taken together, these tests showed positive isolation of PEDV. Using the virus plaque purification cloning methods established in the test, the purified PEDV large plaque and small plaque were obtained, and the large plaque and small plaque titers were measured with significant difference. These results provide potential for the application of PEDV on the basis of the biological features of isolated virus.
Animals ; Cell Culture Techniques ; China ; epidemiology ; Coronavirus Infections ; epidemiology ; veterinary ; virology ; Epidemics ; Feces ; virology ; Porcine epidemic diarrhea virus ; genetics ; growth & development ; isolation & purification ; Swine ; Swine Diseases ; epidemiology ; virology ; Virus Cultivation

OBJECTIVETo assess the results of multi-technique with intact canal and scute rebuild operation for middle ear cholesteatoma.

METHODSAnalysis of 113 cases (males 60, females 53, with mean 42.7 years old), of which with double ears of 11 cases, and totally 124 ears (11 ears for one review, an ear for third time review, and totally 137 operations) was retrospectively performed. Ten cases less than 19 years old were operated from September 2001 to January 2006 for middle ear cholesteatoma as well as ossiculoplasty with intact canal and scute rebuild etc multi-technique. All cases were followed up for a mean (36.58 +/- 20.47) months.

RESULTSThere were 118 ears (95.2%) without cholesteatoma recurrence. Six ears were found cholesteatoma recurrence. Other 6 post-operation ears with drum re-perforation and hearing drop were re-operated. The pre-operation air-bone gap (ABG, average of 0.5, 1, 2 kHz) of 124 ears, with in initial 112 ears, was (33. 61 +/- 12.35) dB, while that of post-operation ABG was (13.58 +/- 9. 27) dB, by partnership t test, t = 18.35, P < 0.01. The pre-operation ABG in reviewed 12 ears at the first pre-operation was (38.83 +/- 12.43) dB, but post-operation ABG (10.38 +/- 8.99) dB, by partnership t test, t = 5.38, P = 0.00022. The ABG closure to within 20 dB was tested in 100 ears while ABG over 20 dB but air conduct (AC) in 40 dB was in 6 ears, and then as succeed was in 106 ears (85.5%). ABG closure within 10 dB was in 50 ears and though ABG over 10 dB but ABG reduced over 30 dB was in 9 ears (59/124, 47.6%) as which as best result. Bone conduct (BC) threshold was tested of 21 ears (16.93%) with raise 1-9 dB, of 42 ears (33.9%) with no change and of 61 ears (49.2%) with descend 1-28 dB. Of 10 cases less than 19 years old post-operation ABG was (8.80 +/- 5.27) dB, which indicated that all cases was as succeed.

CONCLUSIONSUsing intact canal and scute rebuild etc multi-technique a normal canal, hearing improvement and very low recurrent could be gained for middle ear cholesteatoma, especially in juvenile and children. The multi-technique should be almost no damage for hearing bone conduct.

Adolescent ; Adult ; Aged ; Child ; Cholesteatoma, Middle Ear ; surgery ; Female ; Humans ; Male ; Middle Aged ; Otologic Surgical Procedures ; methods ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo observe the effect of Tongsaimai (TSM) tablets in treating foot trauma of diabetic foot (DF) model rats, and discuss its potential mechanism.

METHODMale SD rats were selected to duplicate the diabetic foot ulcer model and randomly divided into the blank control group, the model group, the metformin treatment group, and TSM 12.44, 6.22, 3.11 g x kg(-1) groups (n = 10). The healing of ulcer wounds were observed on day 1, 4, 8, 13 and 18. After 18 days, a histopathologic examination was conducted for ulcer tissues. The contents of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected by hydroxylamine and TBA methods. The content of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were determined with the radioimmunoassay. The immunohistochemical method was used to observe the expression of vascular endothelial growth factor (VEGF) in ulcer tissues and the number of capillary vessels.

RESULTTSM could alleviate the pathological changes of diabetic foot rats, accelerate the ulcer healing on 4, 8, 13, 18 d, reduce MDA, IL-6, TNF-alpha, VEGF content in rat serum at 18 d (after the rehabilitation period), and enhance the SOD content. Specifically, the TSM 12.44 g x kg(-1) group showed significant differences compared with the model group (P < 0.05, P < 0.01). At 18 d after the treatment (the late rehabilitation period), the VEGF expression of TSM 12.44, 6.22 g x kg(-1) groups and the number of blood capillaries of the TSM 12.44 g x kg(-1) group were significantly lower than that of the model group (P < 0.05, P < 0.01).

CONCLUSIONTSM could promote the foot wound healing of DF model rats, reduce MDA, IL-6 and TNF-alpha levels in serum, increase the SOD content and decrease the VEGF expression and the number of blood capillaries in the late rehabilitation period. Its action mechanism may be related to the inhibition of oxidative stress injury and the inflammatory cell infiltration.

Animals ; Diabetic Foot ; drug therapy ; genetics ; metabolism ; physiopathology ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Interleukin-6 ; genetics ; metabolism ; Male ; Malondialdehyde ; metabolism ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; genetics ; metabolism ; Tablets ; administration & dosage ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Wound Healing ; drug effects

Objective To investigate preventive and therapeutic effects ofAngong Niuhuang Pills (ANP) on cerebrovascular diseases in Zebrafish models.Methods Zebrafish cerebral hemorrhage was induced by treatment with simvastatin,thrombosis by arachidonic acid,blood vessel loss by simvastatin,and Alzheimer's disease (AD) by A1C13.Zebrafish models were treated with ANP by direct drug soaking at various concentrations.Results ANP had a significantly preventive effect on Zebrafish cerebral hemorrhage with an efficacy of 11％,41％,and 48％ at concentrations of 27.8,83.3,and 250 μg/mL,respectively,and the incidence of cerebral hemorrhage in 83.3 and 250 g/mL groups decreased significantly compared with model group (P ＜ 0.001).Compared with model group,the red cell staining intensity of Zebrafish in 333 and 1 000 g/mL groups increased significantly (P ＜ 0.001),and the thrombus prevention rates of 111,333,and 1000 g/mL groups were 5％,33％,and 64％ respectively.Compared with model group,the intestinal vascular area of Zebrafish in ANP 11.1,33.3,and 100.0 μg/mL groups increased significantly (P ＜ 0.001),and the rate of angiogenesis was 21％,25％ and 26％ respectively.ANP promoted dyskinesia recovery by 48％,88％,and 78％,and improved response efficiency by 15％,82％ and 75％,respectively in AD zebrafish at concentrations of 111,333,and 1000 μg/mL.Conclusions Our results in this study support ANP as a preventive and therapeutic medicine for cerebral hemorrhage,cerebral ischemia,thrombosis and AD.