An internal amplification control, which could be co-amplified with the invA target gene of Salmonella in the PCR system, was constructed in order to indicate possible PCR inhibitors derived from food samples. Specificity of this PCR system was tested with 9 Salmonella strains and 15 non-Salmonella strains, and the results showed that there was a 374 bp amplicon resulted from all Salmonella strains, while only a 513 bp IAC amplicon appeared after the amplification for all non-Salmonella strains. The detection sensitivity of this PCR system was 12.8 fg/?L for purified target DNA, and the detection limit for artificially inoculated milks was 8 cfu /25g if they were enriched for 8h in buffered peptone water. Salmonella in 80 samples of seriously contaminated milks was detected by the PCR method developed in this study, and the experiments demonstrated that it could successfully eliminate false-negative results.

BACKGROUND: Direct observation of infarcted microvaseular perfusion after cerebral infarction is the most immediate evidence in evaluation of therapeutic effects.OBJECTIVE: To observe dynamic changes in flowing quality and field in cerebral ischemic area in rats so as to qualitatively and quantitatively analyze the microvascular state in pia matral encephali ischemic area and the changes of associated leukocytic flowing velocity and flowing morphology.DESIGN: Randomized controlled experiment was designed.SETTING: Department of Acupuncture and Moxibustion of General Hospital Chinese PLA.MATERIALS: The experiment was performed in Research Room of Microcirculation of General Hospital Chinese PLA, in which, 120 female Wis tar rats of 2-month old were employed and randomized into acupuncture group (36 rats), model group (36 rats), sham-operation group (36 rats) and normal group (12 rats). The first 3 groups were subdivided in 3 hours, 6hours and 24 hours groups according to the prescriptive time divisions, 12rats in each group.METHODS: In acupuncture and model groups, after abdominal anesthesia with pentobarbital sodium, the cranium was opened and the occlusion of meddle cerebral artery was performed in cerebrum with heat-coagulation method. In sham-operation group, anesthesia and cranium opening were performed, but the meddle cerebral artery was not occluded. In normal group, no any management was performed. In acupuncture group, 60 minutes after occlusion of meddle cerebral artery in cerebrum, needling was given on Renzhong (GV 26) and Neiguan (PC 6) with electric stimulation of disperse-dense wave, 4-10 Hz frequency and 0.4 V of intensity, lasting for 5 minutes. Afterwards, pecking-acupuncture technique was done on Renzhong (GV 26) to achieve strong stimulation for 10 seconds. In shamoperation and model groups, the rats were in process of grasping and fixation, but acupuncture was not applied. Vascular endotheliocyte fluorescence staining and leukocyte fluorescence tracing method were applied, in combination with microscopic video system and computer imaging analyzing system, to observe in dynamic and quantitatively the influences of acupuncture on pia matral encephali microvascular morphology, density and blood flowing velocity at 3 hours, 6 hours and 24 hours after cerebral occlusion of meddle cerebral artery.morphology, density and blood flowing state at various time divisions observed directly microscopically and vascular endotheliocyte morphology and the state of brain tissue which near the microvascular after infusion with velocity in infarcted area at various time divisions measured in quantity in each group.microvascular endotheliocytes were colored worse and more fluorescent exudation appeared. Those were better remarkably in acupuncture group ty: That in model group was lower than normal group. That at 3 hours, 6hours and 24 hours in acupuncture group was higher remarkably than model group [(6.92±0.42), (3.25±0.52) pces/visual field; (7.61 ±0.51),(3.68±0.32) pces/visual field; (8.24±0.72), (5.49±1.2) pces/visual leukocyte velocity: That in model group was lower remarkably in normal group. That at 3 hours, 6 hours and 24 hours in acupuncture group was higher remarkably than model group [(1 193±358), (600±261) μm/s;(1 112±267), (517±115) μm/s, (1 766±293), (611±291) μm/s, q=4.608.28, P ＜ 0.01].CONCLUSION: At early stage of cerebral infarction, broken blood flow is severe in artery, vein and capillary in ischemic area; the numbers of opened vessels are few, flow rate is slow down and cerebral perfusion is unsatisfactory. Acupuncture amaliorate the microcirculation perfusion state in ischemic area, increases the numbers of opened vessels and improves flowing morphology of blood cell.

Objective To compare the effect of electroacupuncture at head points and body points on glucose metabolism of the cerebral motor function regions in normal subjects.Methods To observe the change of glucose metabolism of cerebral motor area in normal subjects between before and after acupuncture during the movement by PET.Talairach coordinates(Atlas of brain) and statistical parametric mapping(SPM) software were used to deal with the acquired imaging data.Results ① Acupuncture at Baihui(GV20) and left Qubin(GB7) could increase metabolism of glucose in bilateral Lps and precuneus,the activation of left area in the brain being more significant.②Acupuncture at right Quchi(LI11) and Zusanli(ST36) change metabolism of glucose in left gyrus precentralis,right loblus paracentralis,right gyrus frontalis medialis,both cerebellums and both putarnens.Conclusion All acupoints can change glucose metabolism in cerebral structures related to motor function in the bilateral cerebral hemispheres,different acupoints active different motor areas.The function of acupuncture is a kind of comprehensive regulative process.

OBJECTIVETo explore protective effects of acupuncture at "Zusanli" (ST 36) on cerebral tissue in rats with sepsis.

METHODSCecal ligation and puncture (CLP) was applied to duplicate the rat model of sepsis. According to random number table, thirty SD rats were divided into a sepsis model group (group A), a sepsis model plus electroacupuncture (EA) group (group B), and a sepsis model plus non-acupoint EA group (group C), ten rats in each one. EA with the same frequency and intensity at "Zusanli" (ST 36) and non-acupoint (0.5 cm laterally to "Zusanli") for 30 min was applied in the group B and group C, respectively. No treatment was given in the goup A. 6 hours after CLP, blood was acquired from abdominal aorta to measure the levels of neuron specific enolase (NSE). Then the rats were sacrificed by abdominal aorta exsanguination to take their cerebral tissue for measuring the levels of tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6).

RESULTSSix hours after CLP, the level of NSE was (3.51 +/- 0.39) ng/mL in group B, which was significantly lower than (7.72 +/- 0.64) ng/mL in group A (P<0.05). The level of NSE was (8.02 +/- 0.72) ng/mL in the group C, which had no statistical significance with group A (P>0.05). The levels of TNF-alpha, IL-6 in cerebral tissue in group B were significantly lower than that of group A and C (all P>0.05).

CONCLUSIONEA at "Zusanli" (ST 36) has certain protective effect on septic rat's brain, which has some relationship with decreasing levels of cerebral tissue proinflammatory cytokine and plasma NSE. EA at non-acupoint has no the same action.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Humans ; Interleukin-6 ; immunology ; Male ; Phosphopyruvate Hydratase ; immunology ; Rats ; Rats, Sprague-Dawley ; Sepsis ; enzymology ; immunology ; therapy ; Tumor Necrosis Factor-alpha ; immunology

Objective Analysing the CT manifestations of the crura of diaphragm space occupying lesions and disucssing the diagnosis and differential diagnosis of lesions.Methods 56 cases of the crura of diaphragm space occupying lesions were observed retrospectively,these lesions were confirmed by clinic and pathology.Results The lesions of crura of diaphragm space followed up:15 cases of spinal metastases;5 cases of spinal tuberculosis;27 cases of lymphadenia;1 case of abdominal aorta false aneurysm;3 cases of azygos vein and semiazygos vein dilatory;1 case of taint caused by esophageal perforation;4 cases of invaded by tumour directly. Conclusion We should recognize the nodes and space of crura of diaphragm correctly,that is helpful to recognize the occupying lesions of crura of diaphragm space properly.

OBJECTIVETo study the protective effect of Ligustrazine Injection (LI) against cisplatin-induced ototoxicity and to explore its mechanism.

METHODSThirty healthy adult guinea pigs were randomly divided into three groups, 10 in each group, i.e., the normal control group, the cisplatin group, and the LI group. Guinea pigs in the normal control group were intraperitoneally injected with normal saline at 3 mL/kg for 7 consecutive days. Those in the cisplatin group were intraperitoneally injected with cisplatin at 3 mg/kg for 7 consecutive days. Those in the LI group were intraperitoneally injected with LI at 140 mg/kg for 7 days, but cisplatin (3 mg/kg) was intraperitoneally injected from the opposite side starting from the 4th day. Brainstem auditory evoked potential (BAEP) was performed in all animals before and after injection. All animals were sacrificed after the final testing under anesthesia and their cochleas collected. Half the cochleas of each group were collected for silver nitrate staining of cochlear basilar membrane stretched. The other half the cochleas of each group made into paraffin sections to observe the apoptosis of cochlea cells by TUNEL method and the expression levels of c-Jun detected by immunohistochemistry.

RESULTSThere was no statistical difference in the difference of BAEP threshold among the 3 groups before injection (P > 0.05), but the BAEP threshold increased in the cisplatin group and the LI group (P < 0.05). Besides, it was higher in the cisplatin group (P < 0.05). In the cisplatin group, most hair cells were missing, spiral ganglion cells obviously decreased, more TUNEL positive cells occurred, and the expression of c-Jun was stronger. But the aforesaid impairment in the LI group was obviously lessened (P < 0.05).

CONCLUSIONSLI showed certain antagonist effect on cisplatin-induced ototoxicity. Its mechanism might be associated with scavenging oxygen radicals of the cochlea tissue, improving the microcirculation, and fighting against apoptosis.

Animals ; Apoptosis ; drug effects ; Cisplatin ; toxicity ; Cochlea ; drug effects ; metabolism ; pathology ; Evoked Potentials, Auditory, Brain Stem ; Guinea Pigs ; Pyrazines ; pharmacology ; Reactive Oxygen Species ; metabolism

Objective To demonstrate the clinical outcome of bilateral gluteus maximus musculocutaneous flap in conjunction with continuous postoperative negative pressure wound therapy in reconstruction of sacral soft tissue defects.Methods From January, 2008 to April, 2013, 18 patients (8 males and 10 females, aged from 34 to 78 years old) with full-thickness sacral soft tissue defects were treated.The size of the defects after initial debridement ranged from 3.0 cm × 2.0 cm to 18.0 cm × 14.0 cm, with the exposure of sacrum or ligament.Bilateral gluteus maximus musculocutaneous flap were applied in all the patients.Two drainage tubes were placed on each side of the flaps during the surgery and suck for 10 to 12 consecutive days after the operation.Results The size of the harvested flaps ranged from 12.0 cm × 8.0 cm to 18.0 cm × 12.0 cm, and all the donor sites of the flaps were closed with primary suture.Thirty-five flaps in 17 patients survived without any complication.Partial necrosis of one flap was found in 1 patient and managed successfully with conservative dressing change.Fourteen patients were followed-up ranged from 8 months to 2.5 years (mean follow-up was 18 months).Color and texture of the flaps were satisfactory and no recurrence of sacral defect was noted.Conclusion Bilateral gluteus maximus musculocutaneous flap in conjunction with continuous postoperative negative pressure wound therapy may serve as a useful option for fullthickness sacral soft tissue defects.

Objective To investigate the effects of octreotide on the apoptosis of human hepatic stellate cells (HSCs) and expression of Bcl-2/Bax in HSCs,and to reveal the mechanism underlying octreotide against hepatic fibrosis. Methods HSCs lines (HSC-LX2) were incubated with different concentrations of octreotide for 24 and 48 hours. Cell apoptosis was evaluated by Fitc-tunel fluorescence staining. Bcl-2 and Bax protein exoression in HSC-LX2 was detected by immunocytochemistry. Meanwhile, Bcl-2 protein of HSC-LX2 were detected by Western blot assay. Results Octreotide could promote the apoptosis of HSC-LX2, and the apoptosis rate was significantly increased with the concentration of octreotide(P < 0.05). The HSC-LX2 were incubated with the same concentration of octreotide for 24 and 48 hours, the cell apoptosis rate of 48-hour octreotide treatment was significantly higher than that of 24-hour octreotide treatment (P < 0.05). The immunocytochemistry result indicated that octreotide could significantly decrease Bcl-2 expression and increase Bax expression in HSC-LX2 (P<0.05); Western blot assay showed that octreotide could also significantly inhibit Bcl-2 expression in HSC-LX2 (P<0.05). Conclusions Octreotide could induce the apoptosis of HSCs in a dose-and time-dependent manner, the mechanism of octreotide inducing HSCs apoptosis might be associated with down-regulation of Bcl-2 and upregulation of Bax in HSC.

Objective To investigate the effect of bronchodilators on dyspnea and pulmonary function in patients with chronic obstruction pulmonary disease (COPD) at different degrees.Methods 50 patients with COPD from January 2014 to January 2015 in pneumology department of Tongchuan City People's Hospital were selected.According to the standard of Global initiative for chronic obstructive lung disease (GOLD), the patients were divided into mild degree of 14 cases, moderate degree of 18 cases and severe degree of 18 case.The changes of Borg score of dyspnea, forced expiratory volume in one second (FEV1), forced vital capacity (FVC), peak expiratory flow (PEF) and inspiratory capacity (IC) after received tiotropium bromide inhalation in each group.Results FEV1, FVC, PEF and IC values significant improved in mild, moderate and severe degree groups after received tiotropium bromide inhalation and the values of improvement rate changed significantly with degrees of disease ( P <0.05 ).The improvement rates of FEV1, FVC and IC were highest in patients at severe degree, improvement rate of PEF was highest in patients at mild degree (P<0.05).After received tiotropium bromide inhalation, the Borg score improved significantly compared with that of pre-treatment in mild, moderate and severe degree groups (P<0.05), the improvement rate of Borg score was highest in severe degree group(P<0.05).There were significantly positive correlations between Borg score and FEV1(r=0.372),FVC(r=0.296),PEF(r=0.284),IC(r=0.704)(all P<0.05).Conclusion Bronchodilator could significantly improve dyspnea and lung function in patients with COPD, and the improvement rate of FEV1, FVC and IC is highest in patients with COPD at server degree, PEF is highest at mild degree and Borg score is highest at server degree, which need the comprehensive analysis of each indicators for reversibility of airflow obstruction.

Purpose To retrospectively analyze and summarize the image features of rare ovarian clear cell carcinoma (OCCC), and to provide basis for clinical diagnosis. Materials and Methods CT and MRI features of 30 cases of OCCC confirmed by pathology were retrospectively analyzed. Sixteen of all the patients underwent pre- and post-contrast CT scan. And 14 patients underwent pre- and post-contrast MRI scans. Results ① CT features: unilateral mass was revealed in 15 cases and bilateral mass was in 1 case. The maximum diameter of the mass ranged from 8 to 23.7 cm, mean (12.86±3.96) cm. One mass was irregular, 4 masses revealed incomplete capsule, and 4 masses had septa in the mass. CT value of cystic part of the mass was 20-30 HU, and which of solid part was 28-53 HU. On post-contrast CT images, the septa and solid component of the mass showed marked enhancement and delay enhancement, while the cystic component showed no enhancement. ② MRI features: Unilateral mass was revealed in 13 cases and bilateral mass was in 1 case. The maximum diameter of the mass ranged from 9.2 to 30.0 cm, mean (14.03±4.72) cm. One mass was irregular, 2 masses revealed incomplete capsule, and 2 masses had septa in the mass. The cystic component showed heterogenous signal intensity on T1WI, and high signal intensity on T2WI. There was no enhancement on post-contrast images. The solid component showed iso-intensity on T1WI, high signal intensity on T2WI, and diffused restricted on diffusion-weighted images. There was markedly enhancement on post-contrast images. ③ Blood supply of the tumor: In 8 cases, the branch of enlarged ipsilateral ovarian artery fed the tumors. In other 16 cases, the masses were surrounded by enlarged ipsilateral ovarian vein. Conclusion The characteristic CT and MRI features of OCCC include: a cystic solid mass with complete capsule; the solid component projects into the cavity, which could be hypervascular and marked enhanced.