Adult ; Aged ; Combined Modality Therapy ; Electric Stimulation Therapy ; Electroacupuncture ; Essential Tremor ; therapy ; Female ; Humans ; Male ; Middle Aged

Objective To explore the etiology and pathogenesis of Hirschsprung disease(HD) and the role of nitric oxide in pathophysiology of HD. Methods The whole amount preparations of dilative and transitional segments were stained by NADPH\|diaphorase histochemistry mothod,and colonic walls were digested by dispase. Then the NOS positive nervous structure were observed by light and scanning electron microscope. Results In dilative segment, under light microscope, ganglion and neuron were bigger and more darkly stained. Intraganglionic neurons distributed mostly in peripheral area of ganglion and basal part of ganglionic fiber bundles. Under scanning electron microscope, the neurons were denser, more nerve fibers from them and connected with each other in all dimensions. And more transverse connected fibers between neurons which were arranged along muscle fibers. Myenteric plexus even connected with the submucosal plexus by nerve fibers which passed through circular muscle layer. In transitional segment, under light microscope, intraganglionic neurons cytoplasm were lightly stained and also variedly. The ganglion and neurons of the segment were smaller, and the fibers from them were thinner and paler than those of the dilative segment. Under scanning electron microscope, neurons' density was lower, the fiber connection between neurons and muscle fibers/neurons was lesser. In the transitional segment, neurons and nerve fibers were distributed linearly along the longitudinal muscle fibers. Conclusion Our results indicated that an intimate relationship existed between the development of Hirschsprung disease and abnormal distribution and metabolism of NOS positive neuron in colonic wall. [

Objective To establish a specific fluorescence quantitative method for determining the mRNA expression of Toll-like receptor3(TLR3)in peripheral blood mononuclear cells(PBMCs).Methods Using the Beacon Designer 2.1 software,specific primers and Taqman-MGB probe were designed.The plasmid pMD18-T-TLR3 was constructed as calibrator and the amplified fragment was obtained by reverse- transcript-PCR(RT-PCR).RNA quantification based on cycle threshold values(Ct)was used to establish the standard curve.According to which,the TLR3 mRNA levels in 30 normal individuals,20 patients with primary biliary cirrhosis(PBC)and 20 ones with chronic liver cirrhosis induced by HBV were calculated automatically by software after the fluorescence of PCR product was detected continuously during amplification.Results The linear detection range of the assay for TLR3 gene and ?-actin was 10~2-10~8(r= -0.9974)and 10~3～10~8(r=-0.9984),respectively.The coefficient of variation of both intra-and inter- assay reproducibility for high concentration sample were 6.7% and 8.7%,respectively,and those for low concentration sample were 12.3% and 14.0%.The TLR3 mRNA expression level ranges from 3.46?10~2- 4.51?10~3 copies/?g RNA,4.92?10~2-1.42?10~4 copies/?g RNA and 2.58?10~2-7.17?10~3 copies/?g RNA for 30 healthy individuals,20 PBC patients and 20 ones with chronic liver cirrhosis induced by HBV, respectively.Conclusion We have successfully set up a FQ-RT-PCR method for detecting TLR3 mRNA, which may be used as an excellent tool for the clinic and basic study on the expression of TLR3 gene.

Decompression, Surgical ; methods ; Humans ; Male ; Microsurgery ; Middle Aged ; Vertigo ; surgery ; Vestibulocochlear Nerve ; surgery

ObjectiveTo explore the inductive action of docosapentenoic acid(DPA) on neurite outgrowth in PC12 cells in vitro.MethodsNeurite outgrowth in PC12 cells was examined after the treatment with different concentration of DPA using Motic Zamges Plus software mapping cell image system.Western blot was performed to detect the expression of β Ⅲ-tubulin regulated protein kinase,a neuronal marker as well as ERK and protein kinase B (Akt) phosphorylation.ResultsPC12 cell neurite formation rate was increased in a concentration dependent manner in the induction of DPA,increased by 2.4% (DPA 10 μg/ml,P>0.05),18.6% (DPA 30 μg/ml,P<0.05) and 25.0% (DPA 50 μg/ml,P<0.05) compared with that in the control group.DPA promoted the expression of β Ⅲ-tubulin (P<0.05) and the phosphorylation level of ERK and Akt (P<0.05,P<0.01).ConclusionDPA promotes PC12 cell neurites growth and its mechanism may be related to the activation of ERK and Akt signaling pathways.

OBJECTIVETo explore the effect of electro-acupuncture to improve the bladder function after acute spinal cord injury in rats and its possible mechanism.

METHODSSixty healthy adult male SD rats of SPF grade, with body weight of 220 to 250 g, one week after feeding adaptation, were randomly divided into sham operation group, model group, electro-acupuncture group, electro-acupuncture control group with 15 rats in each group. Sham operation group underwent no stimulation, and the moderate damage model of spinal cord injury were made in other three groups according to modified Allens method. The model group were not treated, electro-acupuncture group were treated with electro-acupuncture on Zhibianxue and Shuidaoxue, and electro-acupuncture control group were treated with electro-acupuncture on 0.5 inch next to Zhibianxue and Shuidaoxue. The frequency of 2/100 Hz, current of 1 mA, stimulation time of 15 min, once a day, left and right alternately stimulate every time, for a total of 7 times. The changes of residual urine volume and urine output in rats at the 1st and the 7th days after operation were observed. And 7 d later, the rats were sacrificed and the injured spinal cord were taken out to observe the apoptosis, and to detect the changes of Bcl-2, Bax, Bad content.

RESULTSAfter modeling,the rats of three groups showed different bladder dysfunction. In electro-acupuncture group and electro-acupuncture control group, the residual urine volume of the 7th day after operation was significant lower than the 1st day after operation (P < 0.001), and there was statistically significant difference on the 7th day after operation between two groups (P < 0.001). Compared with model group, the urine output of electro-acupuncture group and electro-acupuncture control group was significantly increased on the 7th day after operation, and there was sig- nificant difference between electro-acupuncture group and electro-acupuncture control group (P < 0.001). Electro-acupuncture can inhibit apoptosis of spinal cord neurons by TUNEL detection. Postoperative at 7 d, the rate of nerve cell apoptosis in electro -acupuncture group and electro-acupuncture control group was significant increased than model group (P < 0.01, P < 0.05), and there was significant difference between electro-acupuncture group and electro-acupuncture control group (P < 0.005). Compared with model group, the positive expression rate of Bax, Bad decreased (P < 0.01, P < 0.05), and Bcl-2 increased (P < 0.01) in electro-acupuncture group and electro-acupuncture control group,there was significant difference between electro-acupuncture group and electro-acupuncture control group (P < 0.01).

CONCLUSIONElectro-acupuncture can obviously promote the repair of acute spinal cord injury,its mechanism may be through increasing Bcl-2, inhibiting the expression of Bax, Bad, which inhibits the apoptosis of spinal cord neurons.

Animals ; Apoptosis ; Electroacupuncture ; Immunohistochemistry ; In Situ Nick-End Labeling ; Male ; Neurons ; cytology ; physiology ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; pathology ; physiopathology ; therapy ; Urinary Bladder ; physiopathology

Objective To evaluate multi-slice three-dimensional CT angiography (MS 3D-CTA) for the follow-up of intracranial aneurysm clipping.Methods MS 3D-CTA of 16 patients with intracranial aneurysm clipping were retrospectively analyzed.The patients were scanned on a 16-slice spiral CT(GE Lightspeed pro).Volume rendering(VR),thin maximum intensity projection(thin MIP) and multi-planar reconstruction (MPR) were employed in image postprocessing in all cases.Results There were 17 clips in the 16 patients with aneurysm clipping.Six clips were located at the posterior communicating artery,5 at the anterior communicating artery,4 at the middle cerebral artery,and the remaining 2 clips were located at the pericallosal artery in 1 patient.There were no abnormalities found in the aneurysm clipping region in 7 cases by MS 3D-CTA.There were residual aneurysm in 2 cases,parent artery stenosis in 4 cases,and artery spasm in 3 cases.There was no parent artery occlusion and clip displacement in all cases.VR showed excellent 3D spacial relations between the clip and parent artery in 12 cases,and showed good relations in 3 cases.The 1 case with 2 clips in the pericallosal artery showed heavy beam-hardening artifacts.The size and shape of aneurysm clips were clearly depicted by MPR and thin MIP,while 3D spacial relation of aneurysm clip and parent artery were poorly showed.Conclusion MS 3D-CTA is a safe and efficient method for the follow-up of intracranialaneurysm clipping.Combined VR with MPR or thin MIP can well reveal postoperative changes after aneurysm clipping.

Objective To evaluate the clinical application of our self-designed ultradistal locking tool in the intramedullary nailing for tibial fractures.Methods From January 2014 to May 2016,175 patients with tibial fracture were treated at our department.They were 119 men and 56 women,from 19 to 73 years of age (average,46.3 years).They were divided into 2 groups according to the different targeting devices used in the intramedullary nailing.Conventional locking tools were used in the 83 patients from January 2014 to January 2015 and our self-designed new ultradistal locking tools in the 92 patients from February 2015 to May 2016.The 2 groups were compared in terms of operation time,frequency of intraoperative fluoroscopy,and successful rate of one-time locking.Results There were no significant differences between the 2 groups in general clinical data(P ＞ 0.05),showing similarities of the 2 groups.The operation time(59.8 ±4.3 min),frequency of intraoperative fluoroscopy(11.0 ± 2.1 times),and rate of one-time successful locking[94.4％ (238/252)] in the ultradistal locking group were significantly better than those in the conventional locking group [73.6 ± 5.3 min,23.0 ± 3.8 times and 85.7％ (180/210),respectively] (P ＜ 0.05).Conclusions Our new ultradistal locking tools are superior to the conventional ones in that they lead to shorter operation time,less intraoperative fluoroscopy and higher successful rate of one-time locking.Additionally,the new locking tools are easy to handle and incur no extra costs.

It's difficult to identify Aucklandiae Radix and Vladimiriae Radix because of their similar composition. In this paper, UPLC method was used to establish their UPLC fingerprint to identify them with the mobile of acetonitrile -0. 05% phosphoric acid water solution by gradient elution at the detection wavelength of 238 nm. Clustering analysis and principal components analysis showed that Vladimiriae Radix was significantly different from Aucklandiae Radix. Eight common peaks and twelve common peaks were defined respectively in Aucklandiae Radix and Vladimiriae Radix herbs by fingerprint analysis. Six of them were identified as syringoside, chlorogenic acid, isochlorogenic acid A, isochlorogenic acid B, costunolide and dehydrocostuslactone by comparing with standard references. There are four peaks in all of Vladimiriae Radix samples and in none of Aucklandiae Radix samples. So UPLC fingerprint can be used to identify these two herbs.
Asteraceae ; chemistry ; classification ; Chromatography, High Pressure Liquid ; Cluster Analysis ; Drugs, Chinese Herbal ; analysis ; chemistry

Objective To observe the effects of chloroquine phosphate on apoptosis of leukemic cell line U937, and investigate whether chloroquine phosphate induces leukemic cell apoptosis by normalizing protein PNAS-2's abnormal subcellular location. Methods Chloroquine phosphate of different concentrations were added into culture fluid of leukemic cell line U937 at logarithmic phase. MTr was used to measure cell proliferation, flow cytometry and laser confocal microscopy were applied to detect cell apoptosis, and immunofluorescence technology was employed to observe the effects of chloroquine phosphate on the changes of subcellular location of protein PNAS-2. Results Apoptosis of leukemic cell line U937 was significantly induced by 50 μg/mL chloroquine phosphate, and subcellular location of protein PNAS-2 was changed. Conclusion Chlorequine phosphate can induce apoptosis of leukemic cell line U937, and the mechanism may be related to the normalization of PNAS-2's abnormal subcellular location in U937 cell line. Chloroquine phosphate has the potential to be used in leukemic therapy.