As the effects of the nitric oxide and nitric oxide synthase have been investigated a lot,the nitric oxide synthase inhibitors were widely explored and become one of the highlights in the cerebral ischemia research.This paper reviewed the effects of nitric oxide synthase inhibitors especially the nNOS inhibitors and iNOS inhibitors on the cerebral ischemia damage.

BACKGROUND:Previous studies have confirmed the presence of bis-(3'-5')-cyclic dimeric guanosine
monophosphate signaling pathway in Streptococcus mutans, which construct the streptococcus mutans gcp gene knockout strains.
OBJECTIVE:To compare the gene expression differences between Streptococcus mutans wild strains and gcp mutant strains, and to screen the biofilm-related genes from them for the fol ow-up study.
METHODS:The total RNA of two kinds of strains were extracted and stained with cy3 and cy5 respectively after reverse transcription. The gene chip was scanned after hybridization and the differential gene were obtained
through the data analysis. The different expression genes were verified by real-time PCR.
RESULTS AND CONCLUSION:Differential genes were mainly relative about glucose metabolism and biofilm formation. We selected two genes for real-time PCR verification. The PCR results were consistent with the
microarray results. After Streptococcus mutans gcp gene knockout, the gene expressions of gcp mutant strains were upregulated and the gene expressions of phosphotransferase system were downregulated, this result
suggested that two different genes were related with the c-di-GMP signal pathway downstream.

The neuropeptide orexins, including orexin A and orexin B, are located exclusively in hypothalamus OX1R and OX2R are receptors of orexin. OX1R is selective to orexin A while OX2R is not. The distributions of orexin neurons in human brain are the same as those of rats. Orexin was closely related with sleep and awakening behavior besides regulating the feeding behavior. Orexin-containing terminals are also found in areas modulating sleep and awakening such as the locus ceruleus, tuberomammillary nucleus, dorsal raphe, and the basal forebrain. In those areas, there are also lots of OX1R and OX2R and their precursors. Orexin neurons are extremely active during rapid wave sleep and awakening. If hypothalamus in some areas is destroyed and orexin neurons are reduced, sleep-awakening dysfunction will appear (for instance: narcolepsy).

Objective:To analyze the risk factors of first-episode hematogenous metastasis in patients with thoracic esophageal squamous cell carcinoma (ESCC) who received non-surgical treatment after radiotherapy and chemotherapy, and its impact on survival and prognosis.Methods:The clinical data of 230 ESCC patients who met the inclusion criteria and received radical radiotherapy in Tengzhou Central People′s Hospital and Affiliated Hospital of Xuzhou Medical University from January 2011 to October 2018 were retrospectively analyzed.Logistic regression analysis and survival were used to analyze the risk factors and prognosis of blood group metastasis after treatment.Results:In 230 patients with thoracic esophageal cancer, 70 cases (30.4%) developed hematogenous metastasis for the first time.Compared with patients without hematogenous metastasis, the median overall survival was 15 months and 20 months (χ 2=7.249, P=0.007), and the median progression free survival was 9 months and 13 months (95% CI was 7.2-10.8 months and 10.8-15.2 months, respectively χ 2=21.664, P<0.001). Logistic multivariate analysis showed that there was significant difference in the occurrence of hematogenous metastasis among different N stages (χ 2=30.764, P<0.001). N stage was an independent factor for judging hematogenous metastasis, and the increased N stage increased the risk of hematogenous metastasis (OR value were 6.000, 12.629 and 48.167, respectively; 95% CI were 1.712-21.025, 3.546-44.976 and 10.848-213.858, respectively; all P<0.05). The overall survival time of patients with concurrent chemoradiotherapy before hematogenous metastasis was longer than that of patients with sequential chemoradiotherapy and radiotherapy alone (χ 2=10.002, P=0.007). Stratified analysis showed that adjuvant chemotherapy after concurrent chemoradiotherapy could prolong the overall survival of patients with N2 and N3 (χ 2=11.025, P=0.001). Conclusion:N staging is an independent factor to judge the hematogenous metastasis.ESCC patients with hematogenous metastasis after chemoradiotherapy have poor prognosis.N2, N3 patients with concurrent chemoradiotherapy after adjuvant chemotherapy have clinical benefits.

Objective To optimize primary cultures techniques of isolating neonatal rat cardiomyocytes and to com?pare three different methods of extractingβ3-adrenergic receptor(β3-AR)membrane protein from cultured neonatal rat car?diomyocytes. Methods TypeⅡcollagen and differential velocity adhesion were used to collect primary cardiomyocytes. To?tal protein method, ultracentrifugation method, extract kit method were used to isolate cardiomyocytesβ3-AR membrane pro?teins. The BCA method was applied for protein quantification. Relative content ofβ3-AR membrane protein and GADPH in the sample were examined by western blot. Results Optimizing culture and isolation skills can produce a great quantity of cardiomyocytes in high concentration.The kit method acquired a higher level of protein concentration(8.26±0.29)g/L than to?tal protein method(5.12±0.47)g/L does than ultracentrifugation method(3.20±0.37)g/L does all of which were with signifi?cant difference(P < 0.05). The concentration of β3-AR membrane protein was higher if obtained by kit method(0.22 ± 0.05)than ultracentrifugation method(0.09 ± 0.03)than total protein method (0.01 ± 0.01) with significant difference(P <0.05). Conclusion optimizing methodology can obtain abundant myocardial cells in high concentraion. The kit method of isolating primary culturedβ3-AR membrane proteins result in improved concentration and specificity of membrane protein.

Objective To compare the advantages and disadvantages of smear immunohistochemistry and biopsy of bone marrow in the detection of non-Hodgkin lymphoma (NHL) infiltration. Methods 60 newly diagnosed patients with NHL were collected. Smear immunohistochemistry and biopsy of bone marrow were applied to detect bone marrow involvement in those patients. The relations between the results of two detection methods and the relevant clinical characteristics, such as age, blood routine examination, lactic dehydrogenase, extensive infiltration, B symptom, performance status, clinical stage and so on were analyzed. Results Positive rate of bone marrow involvement in bone marrow smear immunohistochemistry group was higher than that in bone marrow biopsy group [10.0 % (6/60) vs. 3.3 % (2/60), P=0.008]. When detection on B cell infiltration, positive rates were 6.6 % (4/60) and 3.3 % (2/60) in smear immunohisto chemistry group and biopsy group, respectively (P=0.007), and when detection on T cell infiltration, the positive rates were 3.3%(2/60) and 0 (0/60), respectively. Furthermore, two detection method was not correlated with related clinical characteristics, such as bone marrow biopsy, immunohistochemistry, gender, age, Karnofsky score, B symptom, extranodal involvement, lactic dehydrogenase, thrombocyte, Hb, neutrophils, lymphocyte and staging (all P>0.05). Conclusions The positive rate of bone marrow smear immunohistochemistry is superior to bone marrow biopsy in the detection of bone marrow involvement of NHL. As for the immunophenotyping of NHL, sensitive rate of bone marrow smear immunohistochemistry is also better than that of bone marrow biopsy.

Objective To analyze retrospectively the MR manifestations and differential diagnosis of extraventricular neurocytoma (EVN).Methods 7 patients with EVN confirmed by pathologic examination were performed non-enhanced and contrast-enhanced MR scan,and one of them was also performed DWI.Results 4 tumors were located in cerebra,2 tumors located in cerebellum,involving both cortex and white matter,and 1 tumor was located in the seller region.5 tumors were solid-cystic lesions and 2 tumors were solid lesions.5 lesions were well demarcated and 2 lesions were ill defined.Edema around the lesion was found in 2 cases.The solid part of tumors showed iso-or hyper intensity on T1WI,hypo-or hyper intensity on T2WI.Necrosis and hemorrhage occurred in some cases.The degree of enhancement in solid part ranged from mild enhancement to intense enhancement heterogeneously.The cystic part showed ring like enhancement with a thin wall.Conclusion The manifestations of EVN on MRI have some characteristics.If a solid-cystic lesion was well defined,involving both cortex and white matter,EVN should be considered.

Objective To investigate the detectable rate and correlative factors of behavior problems among Hui nationality left-behind children in Ningxia rural area.Methods 1394 children,between 6 and 15 years old,of which 282 were Hui nationality children and 112 were Hui left-behind children,were selected from six primary schools and two junior middle schools using cluster random sampling method in Yongning,Ningxia.All of them were assessed by the Achenbach 's Child Behavior Checklist (CBCL) and self-designed General Information Questionnaire.Logistic regression method was used to analyze the correlative factors of behavior problems.Results The detectable rate of behavior problems among Hui nationality children between 6 and 15 years old was 31.25％,which was higher than the Han nationality left-behind children' s(17.84％) and Hui nationality nonleft-behind children' s (14.12％).Among the Hui left-behind children,those who between 6 and 11 years old,had been left alone between 5 and 7,and whose parents exchanging ideas with teachers less than one time a month had higher detectable rates (x2 =14.904,6.327,7.904,P ＜ 0.05).Among the Hui ethnic children,the detectable rates of those who between 6 and 11 yeas old were higher than that of the non-left-behind children' s(P ＜ 0.05).Logistic regression analysis showed that Hui ethnic left-behind children between 12 and 15 years old (OR =0.179) and those whose parents contacted with their teachers frequently(OR =0.362) had less possibilities to attach behavior problems.Conclusion The detectable rate of behavior problems among Hui nationality left-behind Children between 6 andl5 years old in Ningxia is higher,and the correlative factors are complex.

Objective To evaluate the effect of sevoflurane anesthesia on activation of gliocytes in aged rat hippocampi.Methods One hundred and four male Wistar rats,weighing 600-650 g,were randomly divided into 2 groups (n =52 each) using a random number table:control group (group C) and sevoflurane group (group S).Group S inhaled 3.6％ sevoflurane for 2 h,and open reduction and internal fixation was performed after tibial fracture was induced.Y-maze test and fear conditioning test were carried out on days 1,3 and 7 after surgery.Six rats of each group were chosen on days 1,3 and 7 after surgery and hippocampi were removed to detect the activation of microglias and astrocytes (by immunofluorescence) and expression of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) (by Western blot).Results Compared with group C,the number of entries into each arm was significantly reduced,the duration of N arm stay was shortened,the percentage of time spent freezing induced by condition and the percentage of time spent freezing induced by context were decreased,and the activation of microglias and astrocytes was increased,and the expression of IL-6 and TNF-α was up-regulated in S groups (P ＜ 0.05).Conclusion Sevoflurane-induced postoperative cognitive dysfunction may be related to enhanced activation of gliocytes in aged rat hippocampi.

Bombyx mori bidensovirus (BmBDV) has been identified as causing chronic densonucleosis in Bombyx mori specifically. The replication mechanism of BmBDV remains unknown. Its genome comprises two single stands DNA (VD1 and VD2). In order to rescue infectious virions in vitro, we obtained the total viral DNA extracted from the BmBDV-infected larvae midguts, subsequently cloned the full-length sequence of BmBDV genome fragments by PCR and constructed recombinant plasmids pMD18T-VD1 and pUC-VD2. The linear genome fragments were obtained by digesting recombinant plasmids with corresponding restriction enzymes, and then collectively transfected BmN cells by the method of liposome-embedding. We determined the replication of the virus gene by PCR with the template of demethylated total DNA extracted from the post-transfect BmN cells. Meanwhile, we collected the total proteins from the post-transfect BmN cells and the larvae midgut of feeding the post-transfect BmN cells to perform Western blotting analysis, and detected the expression of viral genes. Here we firstly confirm that infectious virions can be rescued in BmN cells by linear co-transfect method.
Animals ; Bombyx ; DNA, Viral ; Densovirus ; growth & development ; Larva ; Transfection ; Virion ; Virus Cultivation