The lineage-Actinobacteria class nov.comprises organisms with a DNA base composition which generally is above 50% G+C (with a few exceptions).We set up a method that has been used in isolating Actinobacteria and Actinomycetes. We added 25?g/mL Nalidixic acid and 25?g/mL Aztreonam into isolation media to inhibit the other bacteria and 20?g/mL Benlete to inhibit fungi.We used fluorescent in situ hybridization to identi 1fy Actinobacteria. Using the four probes,PA-1,PA-2,PHGC and PNHGC,we made the identification on the 31 strains of the 56 gram-positive bacteria randomly selected and got 22 positive results,6 negative results and 3 ambiguous results.It was showed that the results of G+C content determination and FISH method were identical.Among 31 strain,there were 24 strains of Actinobacteria,the rate was 77.4%.This proved the isolation and FISH identification methods were effective and reliable.

Objective To investigate the effect of hydrogen sulfide postconditioning on the systolic function of left ventricle during myocardial ischemia-reperfusion (IR) in rats. Methods Part Ⅰ Adult male SD rats weighing 200-250 g were anesthetized with pentobarbital 40 mg/kg and heparin 250 U/kg. Their hearts were excised and perfused in a Langendorff apparatus with K-H solution saturated with 95% O2-5% CO2 at 37 ℃. Forty isolated rat hearts were randomly divided into 5 groups ( n = 8 each) after 20 min of equilibration: control group (group C); IR group; sodium hydrosulfide 1,10, 100 μmol/L postconditioning group (group SP1, SP10, SP100 ).In group Cthe hearts were perfused continuously for another 100 min. In group IR, the hearts were reperfused for 60 min after 40 min ischemia induced by 10 ml/kg ST. Thomas solution. In group SP1 , SP10 and SP100 the hearts were perfused with K-H solution containing sodium hydrosulfide 1, 10, 100 μmol/L for 2 min before reperfusion.LVDP and ± dp/dtmax were recorded at the end of equilibration and reperfusion. Part Ⅱ Cardiomyocytes were isolated from the male SD rats (weighing 200-250 g) and then cultured in CO2 incubator for 4 h. Sixty-four dishes of cultured myocytes were randomly divided into 4 groups( n = 16 each): control group (group C), hypoxia/reoxygenation group (group HR), hydrogen sulfide postconditioning group (group SP) and hypoxia postconditioning group (group HP). Group C were cultured continuously for 2 h. Group HR, SP and HP were exposed to 1 h hypoxia (95%N2-5%CO2 ) followed by 1 h reoxygenation. In group SP 10 μmol/L sodium hydrosulfide was added and the myocytes were then incubated for 2 min before reoxygenation. In group HP the cultured myocytes were expased to 3 min reoxygenation followed by 3 min hypoxia for 3 times before the 1 h reoxygenation. Mitochondrial membrane potential and F-actin expression were determined. Results Part Ⅰ Compared with group C, LVDP and ± dp/dtmax were significantly decreased at the end of reperfusion in group IR (P ＜ 0.05), while no significant difference was found in group SP1 , SP10 and SP100(P ＞0.05). Compared with group IR, LVDP and ± dp/dtmax were significantly increased in group SP ( P ＜ 0.05). There was no significant difference in LVDP and ± dp/dtmax among group SP1, SP10 and SP100(P ＞0.05). Part H Compared with group C, the mitochondrial membrane potential was significantly decreased in group HR and HP, and the expression of F-actin was significantly up-regulated in group HR, SP and HP ( P ＜ 0.05). Compared with group HR, the mitochondrial membrane potential was significantly increased and the expression of F-actin up-regulated in group SP and HP ( P ＜ 0.05 ). There were no significant difference in the mitochondrial membrane potential and expression of F-actin between group SP and HP ( P ＞0.05).Conclusion Hydrogen sulfide postconditioning can improve left ventricular systolic function during IR in rats by stabilizing mitochondrial membrane potential and promoting aggregation of F-actin.

Objective:To provide clinical data for early identification and diagnosis of vascular embolism in children.Methods:We collected and analyzed the clinical data of children with vascular embolism diagnosed at the Children Medical Center of the First Affiliated Hospital of Hunan Normal University from January 2017 to January 2020.Results:A total of 29 children with vascular embolism were included.The male to female ratio was 2.2∶1(20/9); the age range was 1 month to 13 years, and the median age(IQR) was 16 (6-41)months.Among them, 22 cases were diagnosed with venous thrombosis, including 13 children with lower limb venous thrombosis(13/29, 44.8%), and six children with intracranial venous thrombosis(6/29, 20.7%). Arterial embolism was found in six cases, and left atrial appendage thrombosis was found in one case.Severe pneumonia was the most common primary disease(19/29, 65.5%), followed by cardiopulmonary resuscitation(3/29, 10.3%), and Kawasaki disease(3/29, 10.3%). Analysis on the risk factors of vascular embolization diseases, including catheter-related, long-term bed rest, elevated D-dimer, mechanical ventilation, and intravenous hormone administration, showed that 89.2%(25/29)had ≥3 risk factors at the same time, and 82.8%(24/29)had ≥5 risk factors at the same time.Conclusion:In children with vascular thrombotic diseases, deep venous thrombosis, especially lower extremity venous thrombosis, are common.The severe pneumonia is more common in primary disease.Children with multiple risk factors have a higher risk of developing vascular embolism.In clinic, coagulation function should be monitored and local symptoms should be observed for early identification.

From August 2007 to April 2011,hepatocellular carcinoma (HCC) (n =40),paraHCC tissues (n =10),seminoma (n =10) and cavernous hemangioma (n =10) were selected.And the method of immunohistochemical streptavidin-perosidase was applied to detect the protein expression of Nanog.The expression ratios of Nanog were 17/40 (42％),1/10,0/10 and 5/5 in HCC,para-HCC tissues,seminoma and cavernous hemangioma respectively.Its expression showed no significant correlation with the patient gender,age,serum alpha fetoprotein (AFP),hepatitis B surface antigen (HBsAg),differentiation,Child grade and TNM stage ( P ＞ 0.05 ).It may be used as a surface marker of liver cancer stem cell.

OBJECTIVETo study the related risk factors for surgical site infection following Pilon fracture surgery. METH ODS: The data of 561 patients with Pilon fractures treated with open reduction plate osteosynthesis at our institution's trauma centre were collected from January 2006 to December 2012. All the patients were divided into two groups: infection group and non-infection group. In the infection group, there were 23 males and 10 females, ranging in age from 21 to 69 years old, with an average of (45.50±4.40) years old. In the non-infection group, there were 296 males and 232 females, ranging in age from 16 to 76 years old, with an average of (43.50±7.19) years old. The possible risk factors such as age, gender, smoking, diabetes, alcohol abuse, open fractures, compartment syndrome and operative time were studied. The multivariate Logistic regression model was used to analyze the risk, factors.

RESULTSThe infection rate of surgical site after Pilon fracture surgery was 5.88%. There were significant statistical differences between infection group and non-infection group in operative time, open fractures and compartment syndrome. However, multivariate Logistic regression analysis revealed that only operative time was significantly associated with surgical site infection (P=0.005, OR=44.92).

CONCLUSIONOperation time is an independent predictor for post-operative surgical site infection of Pilon fracture treated with open reduction plate osteosynthesis. Though open fracture and compartment syndrome could increase the surgical site infection rate, they could not not be considered as independent predictors.

Adult ; Compartment Syndromes ; complications ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Operative Time ; Risk Factors ; Surgical Wound Infection ; etiology ; Tibial Fractures ; surgery

Objective:To observe the changes in the anatomy of anterior segment before and after phacoemulsification for treatment of angle-closure glaucoma by Pentacam system,so as to assess the efficacy of phacoemulsification combined with artificial lens implantation(ALI)in treatment of early-and middle-stage angle-closure glaucoma.Methods:Forty-two patients with early-and middle-stage angle-closure glaucoma(44 eyes)were included in the present study.The central anterior chamber depth,anterior chamber volume and peripheral anterior chamber angle were examined by Pentacam system pre-operatively and 1 day,1 week and 3 months post operatively.Meanwhile,A ultrasound was used to examine the anterior chamber depth before and 3 months after operation.and the results were compared with those by Pentacam system.Results:Pentacam system could clearly demonstrate the 3-dimensional structures of the anterior eye segment before and after operation.The central anterior chamber depth,anterior chamber volume and anterior chamber angle were increased significantly after operation(P

Aim To investigate the change in neuropeptide Y(NPY) Y1 receptor level in infarcted myocardium tissues of (MI) rats. 　Method MI was induced by ligating the left descending anterior coronary artery (LAD) in the heart of rats. The techniques of reverse transcription polymerase chain reaction (RT-PCR) were used to identify the exist of NPY Y1 receptor mRNA in myocardial tissues. The technique of semi-quantitative PCR wa s used to observe the change of NPY Y1 receptor mRNA level. 　Results NPY Y1 receptor mRNA distributed in the heart tissues of rat. Compared with sham operated rats ,the NPY Y1 receptor mRNA level both in infarction area and non-infarction area was increased significantly after MI fo r 1d and 3d. The NPY Y1 receptor mRNA level in the heart tissues of sham-op erated rats was also significantly increased compared with that in control rats . 　Conclusion These results suggest that MI may result in increase NPY Y1 receptor level in the heart tissues of rats. Stre ss stimulation such as surgery may also increase the NPY Y1 receptor level in the heart.

Objective To investigate the change in GABA receptor-activated current in dorsal root ganglion (DRG) neurons in rats with neuropathic pain. Methods Twenty adult SD rats of both sexes weighing 100-150 g were randomly divided into 2 gorups: sham operation group (group S, n = 5) and neuropathic pain group (group NP, n= 15). Neuropathic pain was induced by ligation of right L5 spinal nerve. The animals were sacrificed at 5 days after operation. The L5 DRG( neurons in group NP and L3-5 DRG neurons in group S were immediately isolated. Whole-cellpatch- clamp technique was used. The extracellular solution contained GABA 100μmol/L.The frequency and amplitude of the GABA-activated current in DRG neurons and the changes in action potential (threshold potential, rheobase and overshoot) and resting potential before and after GABA administration were recorded. Results GABA 100μmol/L induced rapid inactivation of inward current in most neurons. Compared with the baseline before application of GABA, in group S GABA induced depolarization,increased resting potential and decreased amplitude and rheobase of action potential in large and medium DRG neurons, while in group NP GABA increased resting potential but induced no significant change in threshold potential and rheobase and overshoot of action potential. The frequency and amplitude of GABA-activated current and the degree of change in resting potential and rheobase and overshoot of action potential were significantly lower in group NP than in group S.Spontaneous discharge occurred in small DRG neurons in both groups. No GABA-activated current was observed in all DRG neurons with spontaneous discharge. Conclusions Neuropathic pain is induced by decreasing GABA-mediated inhibition signals in large and medium DRG neurons leading to increased excitability of neurons.

Objective To investigate the effect of hypoxic postconditioning and diazoxide postconditioning on the calreticulin expression during hypoxia/reoxygenation (H/R) in rat cardiomyocytes. Methods Primary cultured male SD rat cardiomyocytes were randomly divided into 6 groups (n = 8 each): control group (group C),H/R group; hypoxic postconditioning group (group HP) and diazoxide postconditioning group (group DP). Group C were cultured continuously for 2 h. Group H/R, Hp and DP were exposed to 45 min hypoxia (95% N2-5% CO2)followed by 1 h reoxygenation. In group HP, the cells were subjected to three cycles of 3 min hypoxia at 3 min intervals at the end of 45 min hypoxia before 1 h reoxygenation. In group DP, diazoxide 50 μmol/L was giyen at the end of hypoxia. Caspase-3 activity, calreticulin expression and intracellular free calcium ion concentrations were determined. Results Compared with group C, caspase-3 activity was significantly increased in the other groups,the calreticulin expression was up-regulated in group HP and DP, and the free calcium ion concentrations were increased in group H/R (P ＜ 0.05 or 0.01). Compared with group H/R, caspase-3 activity was significantly decreased in HP and DP, the calreticulin expression was up-regulated and the free calcium ion concentrations were decreased in group HP and DP (P ＜ 0.01). Conclusion Hypoxic postconditioning and diazoxide postconditioning attenuate H/R injury in rat cardiomyocytes through up-regulating the expression of calreticulin and reducing intracellular calcium overload.

Objective To evaluate the efficacy of long-pulsed tunable Nd: YAG laser on capillary hemangioma and to analyse its influencial factors. Methods A retrospective review was conducted on 124 patients with 135 vascular lesions treated with long-pulsed tunable Nd:YAG laser from February, 2007 to January, 2008. Working parameters, including spot size (3-5 mm), pulse duration (10-30 ms) and fluence (120-200 J/cm2) were determined based on the size, color and depth of lesions and adjusted according to the immediate reaction of lesions. The interval of two treatments was 1 month. Results Of the 135 lesions, 100 experienced a more than 95% subsidence, 22 achieved 60%-94% subsidence, 11 got 30% -59% subsidence, and 2 reached less than 30% subsidence. The cure rate and effective rate were 74.07% and 90.37% respectively. Smaller lesions showed a better response to the treatment than larger ones (P<0.0001), whereas the sites and depth of lesions had no effect on the efficacy (both P>0.05). Atrophy scar occurred in 4 (2.96%) lesions, and blisters in 12 (8.89%) lesions after the treatment. No purpura or pigmentary changes occurred. Conclusion The long-pulsed tunable Nd:YAG laser is an effective treatmnt for capillary hemangioma with fast action and few side reactions.