Objective To investigate the therapeutic effects of Saussurea involucrate injection in severe acute pancreatitis (SAP) in rats.Methods A total of 80 healthy Sprague-Dawley rats were divided into eight groups:SAP group (three hours、48 hours),Saussurea involucrate treated group (three hours、48 hours),ulinastatin control group (three hours、48 hours) and sham operation group (three hours、48 hours),10 rats in each group.After modeling,the rats of SAP group were regularly feeded and the rats of other three group were treated with Saussurea involucrate injection (1.04 mL/kg) intraperitoneal injection,ulinastatin 10 000 U/L tail vein injection,and saline femoral vein injection,respectively and injected every 12 hours.At three hours and 48 hours after treated,blood and pancreatic tissue samples were obtained.The mortality rate,serum amylase level and pathological changes of the pancreas of each group were observed.Serum tumor necrosis factor alpha (TNF-α),interleukin (IL)-6 and IL-10 levels were detected by enzyme linked immunosorbent assay (ELISA).The content of malondialdehyde (MDA) in pancreatic tissues was determined by chemical colorimetry.The level of TNF-α mRNA,IL 6 mRNA and IL-10 mRNA in pancreatic tissues were measured with reverse trascription-polymerase chain reaction (RT-PCR).The activity of nuclear factor kappa B p65 (NF-κB p65) in the pancreatic tissue was tested by immunohistochemistry.Single factor analysis of variance was used to compare multiple groups,and the least significant difference (LSD) method was used in the multiple comparisons between groups.Fisher's exact probability method was performed for rates comparison.Results At 48 hours,there was no statistically significant difference in mortality rate among Saussurea involucrate treated group,SAP group and ulinastatin groups (all P＞0.05).At 48 hours,the histopathology score (8.13 ± 0.64),levels of serum amylase ((2 597.0±214.0) U/L),TNF-α ((254.4±11.6) ng/L),IL-6 ((441.4±14.6) ng/L),levels of pancreatic tissues MDA ((311.0±10.6) mmol/L),TNF-α mRNA(2.04±0.08),IL-6 mRNA (1.77±0.04)and activity of NF-κB p65 ((25.90±2.90)％) of Saussurea involucrate treated group were all lower than those of SAP group (11.40±0.89,(4 780.0±101.0) U/L,(396.0±7.4) ng/L,(664.4± 7.6) ng/L,(418.0± 10.6) mmol/L,2.94±0.03,2.63±0.08 and (51.60±5.27) ％;however level of serum IL-10 ((133.5±6.9) ng/L vs (95.1±5.2) ng/L) and IL-10 mRNA of the pancreatic tissue (1.38±0.06 vs 0.85±0.03) significantly increased (F=253.07、441.63、489.40、2 465.00、196.65、477.89、562.79、131.70、560.18、570.04,all P＜0.01).There was no significant differences in all above parameters between Saussurea involucrate treated group and ulinastatin groups (7.56±0.88,(2 607.0±239.0) U/L,(252.2 ±9.2) ng/L,(443.4±9.6) ng/L,(308.4±9.2) mmol/L,2.10±0.12,1.74±0.04,(26.00±3.67)％,(134.5±7.8) ng/L and 1.42±0.06) at 48 hours (all P＞0.05).Conclusion Saussurea involucrate injection can eliminate oxygen free radicals and prevent to xidation,inhibit NF-κB activation,regulate synthesis and release of cytokines,and alleviate pancreatic injury in SAP rats,but it can not decrease mortality.

Objective The clinical manifestations of cerebral infarction caused by acute basilar arterial occlusion are complex.The purpose of this study is to explore the relationship between lesion location and onset symptoms of cerebral infarction caused by acute basilar arterial occlusion.Methods Fifty three patients diagnosed with cerebral infarction caused by acute artery occlusion were collected from Nanjing Stroke Registry.They were hospitalized in Jinling Hospital from January 2007 to July 2016 and were divided into sudden onset group and progressive onset group.Their clinical and digital subtraction angiography data were analyzed retrospectively.Results Middle and distal segment of the basilar artery occlusions were usually found in sudden onset group.Patients in progressive onset group were more likely to present with proximal segment of the basilar artery occlusions.Significant statistical difference was found between two groups (P<0.05).Logistic regression analysis showed that the symptoms of patients with proximal segment basilar artery occlusion tended to be progressive onset, compared with patients affected by distal segment occlusion (OR=14.77,95%CI:1.57-139.00, P=0.019).Conclusion There was significant relationship between lesion location and onset symptoms of cerebral infarction caused by acute basilar arterial occlusion.Early diagnosis and timely treatment may improve clinical prognosis in patients.

The laboratories of National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, have developed a series of experimental method . These method have unique advantages over the national standard method and industry standard method . How to make these method become public products through legal procedures to serve disease prevention and control more extensively is an obligatory task for national virological laboratories. This article explores the establishment, verification, validation, and examination of virological non-standard method under laboratory certification and accreditation conditions through empirical research on the " RT-RAA method for detecting EV71/CA16 and other viruses" .

Objective: To evaluate the effect of intraperitoneally injected dexmedetomidine on abdominal adhesions in rats and the role of cholinergic anti-inflammatory pathway. Methods: Forty clean-grade healthy adult male Sprague-Dawley rats, weighing 220-250 g, were divided into 4 groups (n = 10 each) using a random number table method: sham operation group (Sham group), abdominal adhesion group (AA group), dexmedetomidine group (DEX group) and dexmedetomidine plus methyllycaconitine group (DEX-M group). The rat model of abdominal adhesions was established by cecal friction method.In Sham group, abdominal cavity was only opened and then sutured.Normal saline 2 ml was injected into the abdominal cavity and tail vein in group AA.In DEX and DEX-M groups, normal saline 2 ml and α7 nicotinic acetylcholine receptor antagonist methyllycaconitine 2.4 μg/g (dissolved in 2 ml normal saline) were injected, respectively, and dexmedetomidine 10 μg/kg (dissolved in 2 ml normal saline) was intraperitoneally injected at the same time.The abdominal incision was opened under anesthesia at 7 days after establishing the model to observe the formation of abdominal adhesion, Phillips method was used for scoring, and enzyme-linked immunosorbent assay was used to determine the transforming growth factor-beta1 (TGF-β1) concentrations in ascites and tumor necrosis factor-alpha (TNF-α) concentrations in serum.The rats were then sacrificed, and the caecum tissue and its contralateral peritoneum and adhesion fibrous strips were obtained for examination of the pathological changes with a light microscope. Results: Compared with group Sham, the abdominal adhesion score and serum TNF-α concentrations were significantly increased in AA and DEX-M groups, and the TGF-β1 concentration in ascites was significantly increased in AA, DEX and DEX-M groups (P<0.05). Compared with group AA, the serum TNF-α concentrations and TGF-β1 concentration in ascites were significantly decreased in group DEX-M, and the abdominal adhesion score was significantly decreased (P<0.05), and the pathological changes of caecum tissue, contralateral peritoneum and adhesion fibrous strips were significantly attenuated in group DEX.Compared with group DEX-M, the serum TNF-α concentrations were significantly increased (P<0.05), no significant change was found in TGF-β1 concentration in ascites (P>0.05), and the pathological changes of caecum tissue, contralateral peritoneum and adhesion fibrous strips were accentuated in group DEX. Conclusion Intraperitoneally injected dexmedetomidine can mitigate abdominal adhesions, and the mechanism is related to activating cholinergic anti-inflammatory pathway and reducing systemic inflammatory response in rats.

Objective: To explore the association between digital devices usage and body weight overestimation in children and adolescents aged 7-18, in order to provide a scientific basis for body weight overestimation prevention in children and adolescents. Methods: Based on the data of the Research Special Project for Public Welfare Industry of Health using stratified cluster sampling method in 2012, a tatal of 40 073 children and adolescents from 7 provinces with complete information were chosen. Ordinal multivariable Logistic regression model estimated the association between digital devices usage and body weight overestimation. Results: A total of 4 276(11.8%) students with overestimation of body weight were detected, who spent >300 min/d time in digital devices(5.12%) than others (3.84%)( χ 2=19.14, P <0.01). Univariate analysis showed that students with time spent on digital devices >300 min/d had a higher risk in overestimation of body weight ( OR=1.36,95%CI=1.18-1.57,P <0.01) compared with students who spent on digital devices≤120 min/d. There was still a significant association after confounder adjustment ( OR=1.28, 95%CI= 1.10-1.48,P <0.05). Stratified analysis showed that the association between digital devices usage and overestimation of body weight were only observed in girls, 11-18 years old and non single child( P <0.05). Conclusion The time usage of digital devices is associated with overestimation of body weight in children and adolescents. It may helpful for children and adolescents to prevent overestimation of body weight by reducing time spent on digital devices.

Objective: To clarify the potential pathogen for fever of unknown origin (FUO) in serum samples for which pathogenic agents were hardly identified with conventional exainatins. Methods: Random capturing the nucleic acid of pathogen was performed by utilizing the property of sequence non-dependence of next generation sequencing (NGS), followed by enrichment of nucleic acid with multiple displacement amplification (MDA). After sequencing, metagenomic analysis was applied to the raw data and the phylogenetic tree was built to identify the potential pathogen. Results: The result did not indicate common pathogens for FUO but showed the existence of Torque teno Viurs (TTV). Assembly was carried out to all sequencing reads. The coverage of consensus sequence on reference was calculated. Phylogenetic result indicated that all confidence sequences belonged to 3 genera (α TTV, β TTV and γ TTV). Conclusions The characteristics of genome, phylogenesis of TTV and TTV as signal at immunology level were analyzed and clarified. The possible explanation for detection of TTV in 3 genera may be that TTV itself or non-infectious factors caused the immunosuppression, which finally result e in rise of TTV detection.

Objective To analyze the expression change of PTGFR gene in hip joint tissues of development dysplasia of the hip (DDH) and its correlation with DDH pathogenesis .Methods Eight age-and gender-matched children with DDH (DDH group) and control children (control group) were enrolled for conducting the compaison .The real-time quantitative PCR method and West-ern-blot method were adopted to detect the PTGFR mRNA and protein expression levels .Results PTGFR mRNA expression level in the hip articular capsule and ligamentum teres of the DDH group were significantly decreased compared with those of the control group (t=3 .472 ,2 .887 ,P<0 .05 ,) .The PTGFR protein expression level in the hip articular capsule and ligamentum teres of the DDH group were significantly decreased compared with those of the control group (t=5 .488 ,3 .942 ,P<0 .05) .Conclusion PTG-FR could play an important role in DDH pathogenesis and may be one of DDH pathogenic genes .

To investigate the presence of integrated Epstein-Barr virus (EBV) DNA in the NK/T cell lymphoma (NKTCL) ge-nome and analyze the integration information in the genome of NKTCL cell lines. Methods: PCR and in situ hybridization were used to detect EBV infection in five EBV (+) NK/T samples and four EBV (-) NK/T samples provided by the biobanks of the First Affiliated Hospi-tal of Zhengzhou University. Whole-genome DNA of the samples was sequenced and subjected to bioinformatics analysis. Whole-ge-nome sequence alignment was used to identify the EBV integration sequence. BLAST analysis was used to compare EBV fasta files of the samples and EBV fasta library. CREST software was used to extract softclip reads, filter all paired reads, and enumerate their distri-bution on chromosomes. The integrated genomics viewer (IGV) was used to compare the distribution of reads in partial regions of chromosome. PCR was used to amplify the high-frequency integration region of the EBV DNA. The amplified fragments were sanger se-quenced. Results: EBV DNA and EBER expression were detected in five EBV (+) NK/T samples but not in the four EBV (-) NK/T samples. Sequencing depth, coverage depth, proportion of coverage, and proportion of alignment all met the requirements for subsequent re-search. Sequence alignment revealed that the captured sequences were viral sequences. Filtered reads were most numerous in EBV (+) NKTCL cell line SNK, YTS, and EBV (+) nasal NKTCL tissue. The reads were non-randomly enriched in chromosome 2. EBV DNA inte-gration in the 400 bp region of chr2:30234084-30234483 caused insertion or deletion in the chr2p23.1 site. Conclusions: EBV DNA is highly integrated in the chr2p23.1 site of EBV (+) NKTCL cells and may affect the expression of related genes.

Objective: To investigate the expression and clinical significance of programmed death-ligand 1 (PD-L1), programmed death-ligand 2 (PD-L2), and their receptor programmed cell death protein 1 (PD-1) in EBV-positive T/NK lymphoproliferative disease [Epstein-Barr virus-positive T/natural killer (NK)-cell lymphoproliferative disease, EBV(+)-T/NK-LPD]. Methods: The pathological paraffin-embedded tissues of 17 patients with EBV(+)-T/NK-LPD from the First Affiliated Hospital of Zhengzhou University from January 2013 to December 2017 were collected. These patients include 12 males and 5 females, aged 10-82 years old, the average age being 29 years, 4 people in gradeⅠ, 7 in gradeⅡ, 3 in gradeⅢ, and 3 people with hydroa vacciniforme-like lymphoproliferative disorders. Immunohistochemical SP method was used to detect the expression of PD-1, PD-L1, and PD-L2 in human EBV(+)-T/NK-LPD tissues. The relationship between PD-1, PD-L1, PD-L2 expression, and clinicopathological parameters, pathological grades and prognosis were analyzed by Fisher's exact probabilities and Spearman rank correlation. Result: After statistical analysis, the results showed that in 17 cases of tissue samples, there were 12 cases with positive PD-1 expression, 6 cases with positive PD-L1 expression and 5 cases with positive PD-L2 expression. There was no significant correlation between PD-1 and PD-L2 expression and prognosis (P>0.05). PD-L1 expression showed a positive correlation with prognosis (P<0.05). There was no significant correlation between the expression of PD-L1 and PD-L2 with age, sex, as well as LDH and Ki-67 levels (P>0.05). Moreover, there was no significant correlation of PD-1 and PD-L2 expression with pathological grade (r=0.141, r=-0.149, both P>0.05). However, there was a negative correlation between the PD-L1 expression and pathological grade (r=-0.563), and the correlation between the PD-L1 ex-pression and pathological grade was statistically significant (P<0.05). Conclusions: PD-1, PD-L1, and PD-L2 are abnormally expressed in the pathological tissues of EBV(+)-T/NK-LPD. Although there was no significant correlation between the expression of PD-1 and prognosis or pathological grade, it was significantly higher in EBV+T/NK-LPD. PD-1/PD-Ls associated signaling pathway is expected to be a potential new target for EBV(+)-T/NK-LPD immunotherapy.

ObjectiveTo explore the mechanism of Dihuang Yinzi （DHYZ）in improving astrocyte injury in the brain and regulating energy metabolism and autophagy disorder in Alzheimer's disease （AD） model mice. MethodForty male APP/PS1 transgenic mice aged four months were randomly divided into a model group and a model + DHYZ group （2.5 g·kg^-1）， with 20 mice in each group. Forty C57BL/6J mice with the same background and same age were randomly divided into a control group and a control + DHYZ group （2.5 g·kg^-1）， with 20 mice in each group. The mice in the control group and the model group were administered with an equal volume of sterilized normal saline by gavage， once a day for 150 days. Novel object recognition test and step-down test were performed to evaluate the learning and memory ability of mice. The expression of glial fibrillary acidic protein （GFAP） in astrocytes was detected by immunofluorescence and Western blot. High-performance liquid chromatography （HPLC） was used to detect adenosine triphosphate （ATP）， adenosine diphosphate （ADP）， and adenosine monophosphate （AMP） in brain tissues of mice， and the data obtained were used to calculate energy charge （EC） levels. The phosphorylation levels of liver kinase B1 （LKB1）， adenosine 5′-monophosphate （AMP）-activated protein kinase （AMPK）， UNC-51-like kinase 1 （ULK1）， and mammalian target of rapamycin （mTOR） and the expression levels of autophagy-related proteins Beclin-1， microtuble-associated protein 1 light chain 3 （LC3）-Ⅱ/LC3-Ⅰ， and p62 in mouse brain were measured by Western blot. ResultCompared with the control group， the model group showed decreased novel object recognition index， shortened retention latency， increased error times in the step-down test， up-regulated protein expression of GFAP， decreased content of ATP， ADP， and EC in brain tissues， elevated AMP ， increased levels of p-AMPK， p-LKB1， and p-mTOR， and protein expression of p62 ， and down-regulated p-ULK1 level and protein expression of Beclin-1 and LC3-Ⅱ/LC3-Ⅰ（P<0.01）， while the above experimental indexes were not significantly different in the control + DHYZ group. Compared with the model group， the model + DHYZ group showed increased novel object recognition index（P<0.05）， prolonged retention latency（P<0.01）， decreased error times（P<0.01） in the step-down test， reduced protein expression of GFAP（P<0.05）， increased content of ATP， ADP， and EC in brain tissues （P<0.05， P<0.01）， decreased AMP content（P<0.05）， reduced p-AMPK， p-LKB1， and p-mTOR levels and protein expression of p62， and up-regulated p-ULK1 level and protein expression of Beclin-1 and LC3-Ⅱ/LC3-Ⅰ（P<0.01）. ConclusionBy protecting astrocytes， DHYZ can improve energy metabolism and autophagy disorder in AD mice to improve the learning and memory ability of model mice.