Objective: To construct attenuated salmonella typhimurium haboring an eukaryotic co-expression plasmid encoding TIP30 and human IFN-? gene and observing its effect on the growth of adenoid cystic carcinoma. Methods: The TIP30 and human IFN-? genes were amplified by PCR and inserted into eukaryotic expression vector pCI-neofor the construction of expression plasmids pCI-TIP30 and pCI-IFN, respectively. A co-expression plasmid pCI-TIP30/IFN was constructed by linking TIP30 and human IFN-? gene using the sequence of internal ribosome binding sequence (IRES). Three recombinant expression plasmids were transformed into an attenuated AroA'autotrophic mutant of salmonella typhimurium SL7207, the resultant bacteria were used to infected murine macrophage in vitro and the expressed products were detected by Western blot and ELISA, respectively. Tumor growth was observed by oral administration of the recombinant salmonella typhimuriums to the nude mouse with adenoid cystic carcinoma. Results: Murine macrophage infected with recombinant salmonella transformed with both plasmids pCI-TIP30 and pCI-TIP30/IFN could express TIP30 protein, and murine macrophage infected with recombinant salmonella transformed with pCI-IFN or pCI-TIP30/IFN could secret human IFN-? in the culture supernatant. Attenuated salmonella typhimurium and three constructed recombinant salmonella typhimuriums all had evident inhibition onthe tumor growth in nude mouse with adenoid cystic carcinoma. Conclusion: The recombinant attenuated salmonella typhimuriums haboring plasmid pCI-TIP30, plasmidpCI-IFN and co-expressing plasmidp-CI-TIP30/IFN were successfully constructed, which could inhibit the growth of adenoid cystic carcinoma in nude mouse.

Objective To investigate levels of IL-17 and IL-22 mRNA in the blood of type 2 diabetes(T2DM)patients with the different stages of kidney injury and explore the relationship between the gene expression levels of IL-17,IL-22 and renal lesions in patients with diabetic nephropathy(DM).Methods Subjects included 60 T2DM patients with or without kidney injury and 20 normal controls(NC,n=20).Diabetes patients were divided into 3 groups by level of urinary albumin to creati-nine ratio (ACR):no proteinuria group (NA,ACR<30 mg/g,n=22),microalb uminuria group (MA,30 mg/g>ACR>300 mg/g,n=18)and diabetic nephropathy group (DN,ACR>300 mg/g,n=20).Quantitative Real-Time RT-PCR was used to detect IL-17 and IL-22 mRNA levels.Analysis differences of IL-17 and IL-22 mRNA levels among NA,MA,DN and NC groups.Results The levels of IL-17 and IL-22 mRNA were significantly higher in DN group than that in MA,NA and NC group (P <0.01,respectively),However,there were not significant difference among MA,NA and NC group (P >0.05,re-spectively).Conclusion Levels IL-17 and IL-22 mRNA were increased in blood of T2DM patients with nephropathy.IL-17 and IL-22 may play role in the pathogenicity of diabetic nephropathy.

Objective To detect the protective mechanism of trehalose in tracheal cryopreservation.Methods Inbred male Sprague-Dawley (SD) rats were sacrificed with intraperitoneal injection of ketamine(150mg?kg -1).The tracheas were removed and immersed immediately in the freezing medium of low potassium dextran (LPD) solution only(Group Ⅰ) ,containing with 10% dimethylsulfoxide(DMSO)(Group Ⅱ), containing with 0.15mol?L -1 trehalose (Group Ⅲ),and containing with 10% DMSO and 0.15mol?L -1 trehalose (Group Ⅳ) respectively. A sterile plastic tube containing a 1-cm-long trachea was filled with the freezing medium,sealed,and frozen to -80℃ at rate of -1℃ per minute in a programmable freezer.Then the tube was stored in liquid nitrogen(-196℃) for 20 days. Then the specimen was thawed in a 37℃ water bath and rinsed with physiologic saline solution 10 times.Histologic changes before cryopreservation and after thawing were examined in each group. After the specimens were embedded in paraffin,5-(m-thick sections were stained with hematoxylin and eosin.The epithelium and cartilage was assessed. We also observed Bcl-2 and Bax gene expression by immunohistochemistry. At last, some tracheas(SD) after cryopreservation were thawed and transplanted into the abdominal cavity of Wistar rats. The transplanted tracheas were retrieved and assessed histologically.Results Microscopic findings of the tracheas in Group Ⅲ and Group Ⅳ showed their structure were intact and Bax gene expression was lower in cartilage after cryopreservation(20d) compared with other groups,especially in Group Ⅳ.The tracheas in Group Ⅲ and Group Ⅳ grew well after they were transplanted into cavity of Wistar rats heterotopically,too.There were no significant differences among 4 groups in Bcl-2 gene expression.Conclusion In tracheal cryopreservation the trehalose can protect the trachea by protecting the tracheal cartilage.It is one of the protective mechanism that the trehalose inhibit the Bax gene expression of cartilage cells.The concomitant use of trehalose and DMSO has a synergistic effect.

Akebia trifoliate has been reported to have many pharmacological activities and the roots, petioles and seeds are used to different symptoms. However, the structure and anatomy of its seeds was almost not reported until now. In the present study, we investigated the morphological characters of the fruit and seed, and the anatomical characters of the testa, micropyle, embryo and endosperm, which could provide evidences for the study on classification, identification and application of A. trifoliate. Our results showed that the testa of A. trifoliate consisted of an epidermic cell layer, the sclerenchyma cells layer, the parenchyma cells layer and an innermost pigment layer. At the micropylar region, the outermost epidermal cells were specialized the white caruncle-like structure and the testa included a lot of lignified tissues. Endosperm comprises two layer cells. Outermost yellowish-brown layer cells contains lots of fat droplets, and innermost white layer cells contains lots of aleurone grains and crystalloids.
Germination ; Magnoliopsida ; anatomy & histology ; growth & development ; Seeds ; anatomy & histology ; growth & development

High resolution melting (HRM), an important technology for genotyping and mutation scanning, has broad prospects in the authentification of traditional Chinese medicine. This paper selected universal trnH-psbA primers and used HRM to establish a new methods for identification of Akebia herbs. PCR was conduct at the annealing temperature of 58 degrees C and 35 cycles. The range of the DNA template concentration, the primer concentration and the Mg2+ ion concentration were further analyzed. The results showed the Tm values of Caulis Akebiae was (81.84 ± 0.16), (85.28 ± 0.16) degrees C and Caulis Clematidis Armandii was (83.22 ± 0.19) degrees C and Caulis Aristolochiae manshuriensis was (81.67 ± 0.14) degrees C, (84.24 ± 0.10) degrees C with 5-125 mg - L-' DNA template, 0.4 μmol x L(-1) primer, 2.0 mmol x L(-1) Mg2+. This method can achieve the authentification of Akebia herbs and is simple, fast, high-throughput, visual.
Chemistry Techniques, Analytical ; methods ; DNA, Plant ; chemistry ; genetics ; Genotype ; Magnoliopsida ; chemistry ; classification ; genetics ; Phylogeny ; Transition Temperature

How to recognize and diagnose heart diseases, as one of the most frequent issues, is a serious problem. ECG automation recognition is not only the target of medicine, but also the work of computer science. In this paper, the approaches to ECG recognition are reviewed, and their characters are analyzed. With the background of wearable wireless ECG monitor, on the basis of experts' experience in ECG recognition, the imagery thinking-based new method, in which morphology parameters are considered, is presented.
Algorithms ; Electrocardiography ; methods ; Humans ; Pattern Recognition, Automated ; Signal Processing, Computer-Assisted

OBJECTIVETo observe the clinical effect of hemiplegic muscle spasticity treated with acupuncture on different acupoints in combination with rehabilitation.

METHODSNinety cases of post-stroke muscle spasticity were randomized into a corresponding and central axis acupuncture group (group A), a conventional acupuncture group (group B) and a rehabilitation group (group C), 30 cases in each one. In group A, the acupoints on the Governor Vessel were mainly selected, as well as those on Hand-Shaoyang meridian and Foot-Taiyang meridian. In group B, the conventional needling and rehabilitation training were applied in combination, of which, the acupoints were mainly from Hand and Foot-Yangming meridians, associated with the acupoints of Shaoyang meridian. In group C, only rehabilitation training was applied. The assessments according to modified Ashworth scale and CSS score were performed before treatment, after 2 weeks and 4 weeks treatment respectively.

RESULTSThe level of modified Ashworth scale of the flexor of elbow and wrist was reduced apparently after treatment in group A as compared with that before treatment (P < 0.01). The level of modified Ashworth scale of the flexor of elbow was reduced apparently after treatment in group C as compared with that before treatment (P < 0.05). There was significant difference on the level change in modified Ashworth scale for the flexor of elbow between group A and group C after 4 weeks treatment (P < 0.05). CSS score decreased significantly in group A as compared with that before treatment (P < 0.01), the improvement after treatment in group A was apparent as compared with the other two groups (P < 0.01).

CONCLUSIONThe corresponding and central axis acupuncture can improve muscle tone of hemiplegic limb, which is superior to the effect achieved by the conventional acupuncture in combination with rehabilitation training and the simple rehabilitation training.

Acupuncture Points ; Acupuncture Therapy ; Aged ; Aged, 80 and over ; Female ; Hemiplegia ; rehabilitation ; therapy ; Humans ; Male ; Middle Aged ; Muscle Spasticity ; rehabilitation ; therapy ; Treatment Outcome

BACKGROUNDGlaucoma can cause progressive damage to retinal ganglion cells. These cells can be classified as cells projecting to the superior colliculus and melanopsin-containing retinal ganglion cells, which project to the suprachiasmatic nucleus. This study was to investigate the effects of chronic intraocular pressure elevation on melanopsin-containing retinal ganglion cells in rats.

METHODSChronic intraocular pressure elevation was induced in one eye of adult Wistar rats by cauterization of three episcleral veins. Intraocular pressure was measured at different intervals with a rebound tonometer. Superior collicular retinal ganglion cells were retrogradely labeled from the superior colliculus with Fluorogold. Melanopsin-containing retinal ganglion cells were visualized by free-floating immunohistochemistry on whole-mount retinas. The number of labeled superior collicular and melanopsin-containing retinal ganglion cells were counted in the sample areas on flat-mounted retinas.

RESULTSCompared with contralateral control eyes, the numbers of both superior collicular and melanopsin-containing retinal ganglion cells were significantly reduced after 12 weeks of experimental intraocular pressure elevation ((2317.41 +/- 29.96)/mm(2) vs (1815.82 +/- 24.25)/mm(2); (26.20 +/- 2.10)/mm(2) vs (20.62 +/- 1.52)/mm(2), respectively). The extent of cell loss of the two types of retinal ganglion cells was similar. However, no morphologic changes were found in melanopsin-containing retinal ganglion cells.

CONCLUSIONBoth melanopsin-containing and superior collicular retinal ganglion cells were damaged by chronic ocular hypertension, indicating that glaucomatous neural degeneration involves the non-image-forming visual pathway.

Animals ; Disease Models, Animal ; Glaucoma ; pathology ; Intraocular Pressure ; Male ; Rats ; Rats, Wistar ; Retinal Ganglion Cells ; pathology ; Rod Opsins ; analysis

In early 2011, the serious outbreak of porcine pseudorabies virus (PRV) infection suddenly recurred in Henan and neighboring Provinces. To investigate the etiology of massive infection with PRV, 16 800 serum samples, 905 porcine epidemic diarrhea virus (PEDV) back-feeding tissues, and 56 PR gene deleted live vaccines were colleted from January 2011 to May 2013 to detect PRV field infection using a PRV gE antibody test kit. The gE and TK genes of 11 new epidemic PRV strains were sequenced by PCR, and their molecular characteristics were analyzed. Moreover, virus titer determination, protective test against PRV, and vaccine potency testing were performed. The results showed that the detection rate of PRV field infection-positive pig farms was 68.06%, and the overall positive rate of PRV field infection in serum was 38.47%; the positive rates in breeding sows, breeding boars, reserve pigs, and commercial pigs were 40.12%, 30.88%, 54.67%, and 26.52%, respectively. The new epidemic strains were in the same evolutionary branch and belonged to the virulent strain group. Compared with the classical PRV strain, the virulence of new epidemic strains changed a little. The length of gE gene was 1 787 bp, and the length of TK gene was 963 bp. The nucleotide homologies of gE and TK genes to Chinese reference strains were 98.2%-99.8% and 98.90%-99.6%, respectively, and the amino acid homologies were 97.1%-99.8% and 97.5%-99.4%, respectively. Commercial vaccine had a 100% protective effect against the new epidemic strains. The positive rate of PRV field infection was 0% in vaccine and 40.44% in back-feeding tissues. The results confirmed that PRV field infection rates were rising sharply among pigs in Henan and neighboring Provinces after 2011. The main virulence genes of new epidemic PRV strains did not change significantly over the years. PR gene deleted live vaccines had no PRV field infection and could completely resist the attack of new strains. The virus carriage of breeding boars and reserve pigs and the serious PRV field infection in PEDV back-feeding tissues were the main causative factors for massive infection with PRV and epidemic outbreak in Henan and neighboring Provinces from 2011 to 2013.
Amino Acid Sequence ; Animal Feed ; analysis ; virology ; Animals ; China ; epidemiology ; Epidemics ; Female ; Herpesvirus 1, Suid ; chemistry ; classification ; genetics ; isolation & purification ; Male ; Molecular Sequence Data ; Phylogeny ; Pseudorabies ; epidemiology ; virology ; Sequence Alignment ; Sequence Homology, Amino Acid ; Sus scrofa ; Swine ; Swine Diseases ; epidemiology ; virology ; Viral Proteins ; chemistry ; genetics

OBJECTIVETo investigate the long-term effect, safety and tolerability of benazepril in general hypertensive patients.

METHODSWe conducted a three-year community-based postmarketing surveillance on benazepril among 1831 essential hypertensive patients (age range from 35 to 88 years) in Shanghai.

RESULTS74.3% of patients persisted in medication taking and were with optimal compliance in a 3-year-follow-up program. Among those taking medication as prescribed after 3 years, 75.7% of them attained systolic blood pressure (SBP) target level of 140 mm Hg (1 mm Hg = 0.133 kPa), 87.4% attained diastolic blood pressure (DBP) target level of 90 mm Hg, and 71.5% attained total target level of 140/90 mm Hg. The reductions were approaching 15 mm Hg for SBP, 10 mm Hg for DBP, and 5 mm Hg for pulse pressure (PP) during the 3 year period. No serious adverse drug reactions (ADRs) were detected during the 3 years follow-up. Cough was the most common ADR. The cumulative incidence of benazepril related cough was 23.6% in women, significant higher than in men (18.8%).

CONCLUSIONBenazepril was safe and tolerable when applied in hypertensive patients.

Adult ; Aged ; Aged, 80 and over ; Antihypertensive Agents ; adverse effects ; therapeutic use ; Benzazepines ; adverse effects ; therapeutic use ; China ; epidemiology ; Cough ; chemically induced ; epidemiology ; Female ; Humans ; Hypertension ; drug therapy ; epidemiology ; Male ; Middle Aged ; Product Surveillance, Postmarketing