Objectives To investigate neuroprotective effect of melatonin on preterm rats brain damage after hypoxia-ischemia (HI). Methods In this study, 5-day-old Wistar rats were divided into four groups: normal saline group, melatonin group, HI+NS group and HI+melatonin group. HI was conducted by unilateral ligation of the left common carotid artery (ische-mia) and 50 min of hypoxia. Melatonin was injected at a dose of 5 mg/kg intraperitoneally three times:before ischemia, after hy-poxia and 24 h after the second dose. The pups were sacrificed at 24 h, 72 h, and 7 weeks after HI;for galectin-3 cells count at 72 h and 7 weeks;TNF-α, IL-1βprotein were measured in 24 h and 72 h after HI;and fear condition and elevated plus maze were tested in 7 weeks after HI. Results The number of galectin-3-positive cells was lower after melatonin treatment than vehi-cle treatment in 72 h and 7 weeks after HI (all P<0.05). TNF-αprotein and IL-1βprotein both increased at 24 h and 72 h after HI, and reduced after melatonin treatment (all P<0.05). Melatonin treatment improved memory ability and learning ability, re-duced anxiety in 7 weeks after HI. Conclusions Melatonin has long-term and short-term protective effect on developing brain damage after HI.

Objective To resolve the problem of the accuracy and standardization of STR-PCR typing in forensic practice,construct DXS6804 allelic ladder by molecular clonning and apply them in a population study on the Pumi population in Yunnan,China.Methods PCR was used to produce several different allelic fragments of the locus.After cloning the PCR products,the recombinant plasmids were sequenced.Then we denominated them and used them as template for re-amplification to generate the locus standard ladder.Results The sequencing results confirmed that the size and the construction of the inserts were correct.The genetic polymorphisms of this locus in Yunnan Pumi population of China were studied.Two off-ladder alleles of DXS6804 locus were found.Conclusion This method is of high value for forensic DNA typing to construct standard ladders.DXS6804 is robust for genetic research and forensic application.

Objective In this study,the fluorine release behavior of six glass ionomer cements (GIC) samples was investigated in distilled water and artificial saliva.Methods The fluorine release behavior in GIC was measured by gas chromatography.Results Various GIC materials had the maxium fluorine release rate in the beginning,and the rate decreased rapidly.After 56 days,although the fluorine release quantity was in a trend of decreasing,the release process can be retained during a long period.For No3,No5,and No6 sampler,the fluorine release quantity in artificial saliva was lower than distilled water,whereas no significant difference was observed for Nol and No2 materials.The No4 sample produced a larger fluorine release quantity in artificial saliva.It can also be conducted that the immersible time of GIC and the fluorine release quantity met the equation of log ( Y ) =a + b x log ( t ).Conclusion The fluorine release behavior of six GIC matched the logarithmic regulation.

Objective:To investigate surgical indications and operative techniques of intracranial arachnoid cysts(IAC). Methods:Thirty patients with IAC treated in Nanjing General Hospital of Nanjing Command were analyzed retrospectively.Cyst wall removal and communicated cyst with adjacent cistern or subarachnoid space was performed in 18 patients,cyst-peritoneal shunt in 8 patients,simple resection in 4 patients. Results: Follow-up CT scans after surgery showed that all cysts disappear or shrinked.Conclusion: Cyst-peritoneal shunt was indicated in recurrent cysts after resection,senile patients and infants.Microsurgery resection of the cyst is preferred in majority of the patients,removal of the cyst wall and communicating the cyst with arachnoid cistern were key procedures for preventing relapse.

ObjectivesTo study the mechanism of brain damage caused byStaphylococcus epidermidis (SE) in mice. Methods A total of 80 neonatal mice of postnatal day 1 (PND1) were divided into SE group (48 mice), normal saline (NS) group (16 mice) and control group (16 mice). Mice in SE group were intravenously injected with 50 μl SE (108/ml). Mice in NS group were given 50 μl NS. Mice in control group were not intervened. At different time points after SE injection (6 h, 24 h, 72 h, 5 d, 7 d), the CFU of brain, blood, and spleen were calculated. Serial sections of parafifn-embedded brain tissue were used for detection of ionized calcium-binding adaptor moleculor1 (Iba-1) by immunohistochemical staining. The positive cells were calculated. ELISA was used to measure the levels of tumor necrosis factor alpha (TNF-α), interleukin-1β (IL-1β), interleukin-5 (IL-5), interleukin-6 (IL-6) of brain at 6 h and 24 h after SE injection.Results There was no SE in brain in different time points. The CFU was at the highest level at 6 h and then decreased after 24 h in blood and spleen. The Iba-1 positive cells in SE group were signiifcantly increased compared to NS group and control group at 24 h and 72 h (P<0.05). There was no difference of Iba-1 positive cells be-tween 24 h and 72 h after SE injection (P>0.05). The levels of TNF-α, IL-1β, IL-5, and IL-6 were signiifcantly higher in SE group than those in NS and control at 6 h and 24 h (P<0.05). The levels of TNF-α, IL-1β, IL-5, and IL-6 were signiifcantly lower in SE group at 24 h than those in SE group at 6 h (P<0.01).Conclusions It is suggested that cytokines produced by microglias may be the mediators of SE-caused brain damage.

Objectives To study the influence of Staphylococcus epidermidis (SE) on the neonatal mice of different ages. Methods A total of 60 neonatal mice including postnatal day 1(PND1) and postnatal day 3(PND3) were divided into SE group, normal saline (NS) group and control group, with 20 mice each. Mice in SE group were intravenously injected with 50μl SE (108/ml). Mice in NS group were given 50μl NS and mice in control group was not intervened. On postnatal day 14, the brain, liver and spleen obtained from mice were weighted. Serial sections of paraffin-embedded brain tissue were used for the detec-tion of microtubule associated protein-2 (MAP-2) and myelin basic protein (MBP) by immumohistochemical staining, and then the areas and volumes of grey and white matter were calculated. Result The mortality of PND1 mice in SE and NS group was 60.0%and 40.0%, respectively, and there was no difference between two groups (P>0.05). The mortality of PND3 mice in SE and NS group was 10.0%and 0.0%, respectively, and there was no difference between two groups (P>0.05). There were no dif-ferences in body weight, body weight gain, spleen and liver weights and organ coefficient between PND1 and PND3 mice (P>0.05). In PND1 mice, the areas and volumes of grey and white matter were significantly smaller in SE group than those in NS group (P<0.05). However, in PND3 mice, there was no differences in areas and volumes of grey and white matter between SE and NS group (P>0.05). Conclusions SE infection can result in brain injury in PND1 mice, but has no effect on brain tissues of PND3 mice.

Child ; Choristoma ; pathology ; Diagnosis, Differential ; Follow-Up Studies ; Humans ; Male ; Neoplasms, Glandular and Epithelial ; pathology ; Thymus Gland ; pathology ; Thymus Neoplasms ; pathology ; Thyroid Neoplasms ; pathology

Objective To observe the effect of Wnt/β-catenin pathway on proliferation of human umbilical vein endothelial cell (HUVEC) using the glycogen synthase kinase 3β (GSK-3β)-targeting RNAi recombinant adenovirus vector. Methods Homologous recombination and cloning techniques were used to construct RNAi recombinant adenoviral expressive vectors specific to GSK-3β. Then, the adenovirus plasmids was transfected into HEK 293A cells to produce adenovirus and amplify the adenoviral stock. Plaque forming assay was used to titer the adenoviral stock. The GSK-3β and β-catenin protein expressions were detected by Western blot and immunohistochemistry. The proliferation of HUVEC was detected with MTr assay. Results The RNAi adenovirus vectors specific to GSK-3β were successfully produced with high titer. The expression of GSK-3β protein in HUVEC could be down-regulated efficiently by the RNAi adenovirus, along with increased β-catenin protein expression. The proliferation of HUVEC was significantly increased (P < 0.05 or P < 0.01) after infected with GSK-3β RNAi recombinant adenovirus for 3, 5, 7 days. Conclusion RNAi adenovirus is an important tool that can inhibit the expression of GSK-3β efficiently, along with increased β-catenin protein expression. Up-regulating of the Wnt/β-catenin pathway might play an important role in the proliferation of HUVEC.

A promising genetic marker, sag5b, was cloned and expressed and the difference of the genes between highly virulent strain (RH) and less virulent strain(Prugniaud) of Toxoplasma gondii was compared. The PCR-generated product of sag5b was subcloned into T easy vector and plasmid pET28a consecutively. The fusion expression was induced by IPTG and identified by SDS-PAGE and Western blotting. The immunoreactivity of recombinant SAG5B was identical to that of native SAG5B on the membrane of tachyzoites of RH strain. The brains of mice infected with Prugniaud strain of T. gondii were homogenated. Sag1 was successully cloned by PCR from both RH strain tachyzoites and the homogenized brain tissues of mice infected with low virulent strain of Prugniaud,whereas sag5b was only detected in RH strain but not in Prugniaud strain, indicating that sag5b could be used as a genetic marker for differentiation of strain virulence. Expression and vaccination of the virulence-associated gene into mice failed to induce obvious protective immunity against the challenge of RH strain.

Objective To determine the chlorpyrifos in human blood by liquid chromatography-tandemmass spectrometry and to validate its application in poisoning cases. Methods The samples were extracted by a simple one-step protein precipitation procedure. Chromatography was performed on a Capcell Pack C18 mG II column (250 mm×2.0 mm, 5μm) using an isocratic elution of solvent A (0.1% formic acid-water with 2 mmol/L ammoniumacetate) and solvent B (methanol with 2 mmol/L ammoniumacetate) at 5∶95 (V∶V).Results The linearranged from5 to 500ng/mL (r=0.9987).Thelimitofdetection (LOD) and the lower limit of quantification (LLOQ ) were 2 ng/mL and 4 ng/mL , respectively. For this method, the precision and accuracy of intra-day and inter-day were <10% and 97.44%-101.10%, respectively. The re-sults in stability test of long-termfrozen were satisfied. The matrix effect, recovery and process efficien-cy were 64.97%-86.81%, 76.70%-85.52%, and 55.57%-66.58%, respectively. Conclusion This method can provide a rapid approach to chlorpyrifos extraction and determination in toxicological analysis of forensic and clinical treatment.