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.

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 To provide the anatomical data for single-nostril transsphenoidal microsurgery for pituitary adenoma.Methods Microanatomic structures were observed and measured in 20 cadaveric a- dults heads.Results(1)The diaphragmal opering was 5 mm or greater broad in 90%.The mean thick- ness of the diaphragma sellae was (0.18?0.08)mm.(2) A sphenoid sinus with complete pneumatization was named as all-sellae type,its percentage in all specimens was 85%,85% sphenoid sinus had middle sep- tum of which only 18.8% located in the medial sagital section.(3)The meant thickness of the sellar floor was (0.81?0.34) mm,70% of which were not more than 1 mm.Of the sellar floors,that the shape was flatness reach 15%.(4) The relationship between ICA and sphenoid sinus had two aspects,one was to be mutually apart (55%),the other was to communicate (45%),which formers carinae on the lateral wall of the sphe- noid sinus,espeially in the superior surface of posterior part.The relationship between optic nerve and sphe- noid sinus,one was to be mutually apart (27.5%),the other was to communicate (72.5% ),which formers carinae on the lateral wall of the sphenoid sinus,especially in the superior surface of anterior part.(5) Three main types of the intercavernous sinus were the anterior intercavernous sinus,present in 80%,the inferior in- tercavenous sinus (25%),and the posterior intercavenous sinus (15%).Conclusion The knowledge of the anatomy of the sellar region and sphenoid sinus is helpful to safe operation in the single-nostril transsphe- noidal approach for pituitary,adenoma microsurgery.

AlM: To observe the therapeutic effects of improved external dacryocystorhinostomy.
METHODS:Retrospective analysis on 94 patients with monocular chronic dacryocystitis in our hospital from October 2010 to December 2013 were taken the improved external dacryocystorhinostomy. The improved surgery which was based on the traditional surgery including: 1. Nasal packing after anesthesia to relieve the pain and bleeding; 2. Not cut the medial palpebral ligament; 3. Suture the upper membrane of the nasal mucosa only;4. Pipe placement;5. Skin layered hairdressing suture. The patients were follow-up 3mo-2a after operation.
RESULTS: Ninety-three cases of patients cured with completely asymptomatic, no epiphora or mucopurulent secretion flow out from the lacrimal punctum, unobstructed lacrimal irrigation, the efficiency is 99%, 1 case of patient was epiphora, obstructed lacrimal irrigation, 1% was invalid.
CONCLUSlON:The improved external dacryocystorhinostomy is an effective surgical method which is easy to operate with high cure rate and its long term effect is precise.

Objective To understand the iodine deficiency disorders in Xunhua county Qinghai province, and to provide reference values for setting up prevention and control strategy. Methods According to historical data of Xunhua county, 1 township center elementary school and 1 village elementary school were sampled in Qingshui, Baizhuang, Hongqi, Jiezi and Galeng townships. A total of 9 elementary schools were investigated (Caleng has only 1 elementary school). Fourty children aged 8-10 years were checked thyroid size by palpation in every school. Twenty children were chosen to assay their urinary iodine and edible salt iodine in every school. Thirty 5-grade students were surveyed by questionnaire on health education in every school. Ten housewives were inquired prevention knowledge on iodine deficiency disorders and salt iodine level was tested in every village. Results Goiter rate was 7.9%(30/378) in children aged 8-10 years, with Baizhuang and Galeng the highest (11.9%,5/42), and Qingshui the lowest (2.4%, 1/42). The median was 196.5 μg/L in children aged 8-10 years, and urinary iodine level lower than 50 μg/L was accounted for 5.3% (11/208), but that value was 25.0% (5/20)in Galeng. The average qualified rate of using edible salt was 91.4%(159/174), and that rates were 79.0%(15/19), 83.3%(15/18) and 89.5%(17/19) in Chajia, Tuanjie and Minzhu villages, respectively. The average score were 2.8, 2.1 in health education survey of the 5-grade students and housewives. Conclusions After years effort in prevention and control of iodine deficiency disorders, remarkable progress has been made in Xunhua County. The monitoring of iodized salt and the knowledge spreading are still need to be strengthened.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Cluster Analysis ; Diagnosis, Differential ; Female ; Humans ; Liver Diseases ; classification ; epidemiology ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Yang Deficiency ; classification ; epidemiology ; Yin Deficiency ; classification ; epidemiology

AIM:To investigate the ability of a metal complex ammonium tetrathiomolybdate (ATTM) to re-lease H2 S and its cytoprotective effect on an oxidative injury model .METHODS:Released H2 S was absorbed in a reaction flask from ATTM dissolved in the cell medium .Staining with dichlorodihydrofluorescein diacetate or rhodamine 123 fol-lowed by photofluorography was conducted for the observation of reactive oxygen species ( ROS) and mitochondrial mem-brane potential (ΔΨm) levels, respectively.Cell viability and release of lactate dehydrogenase (LDH) from the cells were measured with commercial kits.RESULTS:Similar to another H2S donor GYY4137, ATTM had an ability to release H2S in the cell medium in a dose-dependent manner .Treatment of human skin HaCaT cells with ATTM at concentrations of 25~400 μmol/L didn’ t significantly alter cell viability .Exposure of the cells to ultraviolet rays or a ROS donor H 2 O2 in-creased the intracellular ROS levels .Treatment with 400 μmol/L H2 O2 significantly reduced the viability of HaCaT cells (P<0.01).However, before the treatment with H2O2, pretreatment with ATTM at 100 and 200 μmol/L markedly pre-vented the H2O2-induced cell injury (P<0.01).In addition, the treatment with H2O2 triggeredΔΨm loss (P<0.01) and LDH release from the cells (P<0.01).Prior to suffering from H2O2 injury, the preconditioning with 200 μmol/L ATTM significantly improved ΔΨm levels ( P<0.05 ) and attenuated LDH release from the cells ( P<0.01 ) .CONCLUSION:ATTM is capable of releasing H 2 S and protecting human skin cells against oxidative injury .

Background Allograft rejection is a main cause of failure of penetrating keratoplasty,especially in the patient with high risk of rejection condition.Previous study on allograft rejection mechanism focused on limbal and corneal neovascularization,but these factors did not explain all the phenomena of allograft rejection.Research found that immune cells appeared in iris and ciliary body when rejection occurred,but the relationship between these immune cells and allograft rejection is unclear Objective This study was to evaluate the relationship between diversity of vascular permeability in the iris and ciliary body and allograft rejection after penetrating keratoplasty.Methods Seventy clean eight-week-old BALB/c mice were divided into allogeneic corneal transplantation group (60 mice) and blank control group (10 mice).Allogeneic corneal transplantation was performed with the same age of C57BL/6 mice as donor and BALB/c mice as the recipients.The grafts were examined under the slit lamp microscope and scored based on the criteria of Hegde.The mice were sacrificed and iris and ciliary tissue were obtained 5,10 days and rejection after surgery.Immunohistochemistry and reverse transcription PCR (RT-PCR) was used respectively to detect the expression diversities of occludin,zonula occludens protein-1 (ZO-1),matrix metalloproteinase-9(MMP-9),major histocompatibility complex-Ⅱ (MHC-Ⅱ),and CCR5,CCR7 and their mRNA in iris and ciliary body.Image-J image analysis software was used to calculate the quantity of positive cells on iris wholemount,and absorbance of target genes (A values).The use and care of the experimental animals complied the ARVO Resolution on the Use of Animals in Research.Results The mean survival time of corneal gratts was (17±3) days after operation.The mean score was 0.6 in 5 days and 0.5 in 10 days,and 3.3 in 18 days after operation.Expression of ZO-1 reduced significantly,and that of MMP-9 increased obviously at the time of rejection.MHC Ⅱ + cells were scattered in iris and ciliary body in normal mice,and the number of the positive cells (cells/field) was increased after operation with a peak value when rejection occurred.A significant difference was seen between normal mice and rejection mice (1559.67±350.29 vs.4021.83±495.18) (P=0.000).The expressions of occludin mRNA and ZO-1 mRNA in the iris and ciliary body decreased obviously in the rejection mice.Compared with normal mice,theA value of ZO-1 and occluding were 36.74±3.13 vs.110.11±11.88 and 57.54±3.41 vs.59.90±3.50respectively,with significant differences between them (all P＜0.05).The expressions of MMP-9 mRNA,CCR5 mRNA and CCR7 mRNA in the iris and ciliary body increased gradually with the time lapse after operation and peaked when the rejection appeared.The A value of MMP-9 mRNA,CCR5 mRNA and CCR7 mRNA were significantly higher than those of normal mice (20.29±1.19 vs.2.77±0.85 for MMP-9 mRNA; 35.43±2.56 vs.9.11±0.29 for CCR5 mRNA,and 60.83±0.87 vs.0.89 ±0.95 for CCR7 mRNA) respectively (all P＜0.05).Conclusions The permeability of vascules in the iris and ciliary body increase during the allograft rejection after penetrating keratoplasty.Increased antigen presenting cells were also detected.

BACKGROUND: von Willebrand factor (vWF) is only released from endothelial cells and platelets and is an in vivo and in vitro marker of endothelial injury in septic patients with acute lung injury (ALI). Interleukin-8 (IL-8), as a proinflammatory mediator causing recruitment of inflammatory cells, induces an increase in oxidant stress mediators and makes it as a key parameter for localized inflammation. However, it has not been well established whether the level of serum IL-8 is associated with the severity of lung injury and whether it is a prognosis marker for severe lung contusion. This study was to investigate the expression of plasma vWF and IL-8 and their association with the severity and outcomes of severe pulmonary contusion. METHODS: A total of 63 patients were divided into a severe pulmonary contusion with acute respiratory distress syndrome (ARDS) group and a non-ARDS group, or a survivor group and a non-survivor group, or an injury severity score (ISS) <20 group and an ISS ≥20 group. Another 20 healthy volunteers served as controls. The levels of plasma vWF and serum IL-8 were measured by enzyme-linked immunosorbent assay (ELISA) at 1, 3, 5 and 7 days after injury. The expression patterns of the plasma vWF and serum IL-8 were compared between different groups. RESULTS: The concentrations of plasma vWF and serum IL-8 were significantly increased in all severe pulmonary contusion patients at all time points in comparison with the control group. The concentrations of plasma vWF in patients with ARDS increased during the whole study period, but vWF in patients with non-ARDS increased gradually until day 5 and then decreased at day 7. The concentration of serum IL-8 showed a similar expression pattern in both groups, but the expression increased more significantly in the ARDS group than in the non-ARDS group. Interestingly, both plasma vWF and serum IL-8 levels steadily increased in the non-survivor group. Furthermore, the level of plasma vWF was higher in the ISS≥20 group than in the ISS<20 group. The level of serum IL-8 in the ISS≥20 group was consistently high, while that in the ISS<20 group peaked at day 3 and decreased at day 5. In addition, the level of plasma vWF was positively correlated with platelet count, but negatively correlated with oxygen index. The level of serum IL-8 was positively correlated with white blood cell count and ISS score, and inversely correlated with oxygen index. CONCLUSION: The elevated levels of plasma vWF and serum IL-8 in severe pulmonary contusion patients reflect the severity of pulmonary injury and patients outcomes, suggesting that the plasma vWF and serum IL-8 are sensitive markers for clinical evaluation of the severity of pulmonary injury and predication of patient prognosis.

OBJECTIVETo investigate the effects of electroacupuncture at "Zusanli" (ST 36) on the volume of hepatic blood flow, water ratio and plasma alanine aminotransferase (ALT) in rats with delayed fluid replacement after hemorrhagic shock and to provide the references for electroacupuncture at Zusanli (ST 36) in treating hemorrhagic shock.

METHODSForty SD rats with hemorrhagic shock induced by bloodletting 40% of whole blood volume were randomly divided into a hemorrhage with no treatment (NT) group, an immediate fluid replacement (IFR) group, an electroacupuncture at Zusanli (ST 36) and delayed fluid resuscitation (EA/DFR) group and a sham electroacupuncture and delayed fluid replacement (SEA/DFR) group, 10 rats in each group. No treatment was performed in NT group. IFR group was treated with fluid replacement at 10 minutes after blood loss, and EA/DFR group was treated with electroacupuncture at "Zusanli" (ST 36) at 10 minutes after blood loss, while non-acupoint was punctured in SEA/DFR group. Two EA groups were received delayed fluid replacement at 3 hours after blood loss. The volume of hepatic blood flow and ALT before blood loss and 3 h and 12 h after blood loss, and water ratio 12 h after blood loss were measured.

RESULTSAfter blood loss, all parameters in IFR group and EA/DFR group were improved significantly in contrast with those in NT group (all P < 0.05). There was no significant difference between SEA/DFR group and NT group. Three hours after blood loss, the hepatic blood flow of IFR group was significant higher than those of NT group, EA/DFR group and SEA/DFR group (all P < 0.05), while the plasma ALT of IFR group was significant lower than those of NT group, EA/DFR group and SEA/DFR group (all P < 0.05), and the plasma ALT of EA/DFR group was lower than those of NT group and SEA/DFR group (both P < 0.05), the hepatic blood flow of EA/DFR group showed no significant difference compared with that of SEA/DFR group (P > 0.05). Twelve hours after blood loss, the plasma ALT and the water ratio of EA/DFR group and IFR group were significant lower than those of NT group and SEA/DFR group (all P < 0.05), and the hepatic blood flow of EA/DFR group and IFR group was significant higher than those of NT group and SEA/DFR group (all P < 0.05), while the plasma ALT of IFR group was significant lower than that of EA/DFR group (P < 0.05), and the hepatic blood flow of IFR group was significant higher than that of EA/DFR group (P < 0.05).

CONCLUSIONElectroacupuncture at "Zusanli" (ST 36) has a protective effects for hepatic ischemic injury in rats with delayed fluid replacement after hemorrhagic shock.

Acupuncture Points ; Animals ; Electroacupuncture ; Humans ; Ischemia ; therapy ; Liver ; blood supply ; Male ; Rats ; Rats, Sprague-Dawley ; Shock, Hemorrhagic ; therapy