Objective To investigate the role and underlying mechanism of atorvastatin on hyper-glycemia induced hemorrhagic transformation(HT)in a mouse model of cerebral ischemia.Meth-ods A total of 36 SPF-grade male C57BL/6 mice were randomly divided into sham operation group,HT model group and atorvastatin group,with 12 mice in each group.HE staining was used to observe cerebral hemorrhage,immunofluorescent staining was employed to detect the integrity of blood-brain barrier,and Western blotting was applied to measure the protein expression of IgG,ZO-1,occludin,claduin5,MMP-2 and-9 in ischemic penumbra brain tissues.Results Com-pared with sham operation group,the neurological deficit score,mortality rate,HT incidence,HT grading score,IgG fluorescence intensity,and protein levels of IgG,MMP-2 and-9 were signifi-cantly increased,while the protein levels of ZO-1,occludin and claudin5 were obviously decreased in the HT model group(P＜0.01).Atorvastatin treatment resulted in significantly lower neuro-logical deficit score(2.73±1.19 vs 3.91±0.94),mortality rate(16.7％vs 41.6％),HT incidence(58.3％vs 91.6％),HT grading score(1.00±1.04 vs 2.58±1.13),IgG fluorescence intensity(504.30±105.52 a.u vs 859.91±153.28 a.u),and protein levels of IgG(4.55±1.40 vs 12.06± 3.73),MMP-2(1.87±0.41 vs 2.95±0.68)and-9(1.47±0.24 vs 2.12±0.23)(P＜0.05,P＜0.01),and increased protein levels of ZO-1(1.55±0.20 vs 0.53±0.10),occludin(0.92±0.11 vs 0.35±0.07)and claudin5(0.58±0.04 vs 0.30±0.05)(P＜0.01)when compared with the HT model group.Conclusion Atorvastatin can reduce the permeability of blood-brain barrier by in-hibiting the activation of MMP-2 and MMP-9 and up-regulating the protein levels of ZO-1,occlu-din and claudin5,and thus attenuate hyperglycemia-induced HT.

Gastric cancer with hemorrhage and cerebral infarction is a serious complica-tion with poor prognosis in clinic. Although the incidence rate is extremely low, the fatality and disability rates are very high. In addition, the opposition in treatment between the two complica-tions increases the difficulty of clinical diagnosis and treatment. The authors report the diagnosis and treatment of a gastric cancer patient with hemorrhage and new cerebral infarction, in order to to provide reference for related treatments.

ObjectiveTo systematically review the effect of psychological intervention on anxiety and depression in hearing-impaired children and adolescents. MethodsThe literature was researched from the databases of PubMed, Web of Science, Embase, CNKI, and Wanfang Database on psychological intervention on anxiety and depression for hearing-impaired children and adolescents until July, 2023. After screening, author, country, study object, study design, intervention mode, outcome, and measurement tools were extracted from the literature. ResultsEight articles from three countries were included, including eight randomized controlled trials, involving 404 participants, publishing mainly from 2013 to 2023. The types of psychological intervention included family support education, emotional intelligence training, cognitive behavioral therapy, self-confidence group training, and play therapy, and so on, which improved social phobia, and relieved the levels of anxiety and depression of children and adolescents with hearing impairment. ConclusionPsychological intervention can effectively alleviate the anxiety and depression of children and adolescents with hearing impairment.

Objective:The investigation and research on the application status of Hepatic Venous Pressure Gradient (HVPG) is very important to understand the real situation and future development of this technology in China.Methods:This study comprehensively investigated the basic situation of HVPG technology in China, including hospital distribution, hospital level, annual number of cases, catheters used, average cost, indications and existing problems.Results:According to the survey, there were 70 hospitals in China carrying out HVPG technology in 2021, distributed in 28 provinces (autonomous regions and municipalities directly under the central Government). A total of 4 398 cases of HVPG were performed in all the surveyed hospitals in 2021, of which 2 291 cases (52.1%) were tested by HVPG alone. The average cost of HVPG detection was (5 617.2±2 079.4) yuan. 96.3% of the teams completed HVPG detection with balloon method, and most of the teams used thrombectomy balloon catheter (80.3%).Conclusion:Through this investigation, the status of domestic clinical application of HVPG has been clarified, and it has been confirmed that many domestic medical institutions have mastered this technology, but it still needs to continue to promote and popularize HVPG technology in the future.

With the rapid development of transgenic technology, the safety of genetically modified products has received extensive attention. Certified reference materials for the detection of genetically modified organisms play important roles in ensuring comparability and traceability of the qualitative and quantitative detection of genetically modified products. However, the development of protein reference materials is relatively slow, and one of the difficulties is the preparation of protein candidates with high purity. The cry1Ah1 gene of Bacillus thuringiensis has been used for the development of transgenic insect-resistant crops because of its excellent insecticidal activity against lepidopteran pests such as Asian corn borer, and has obtained transgenic lines with good insect resistance traits. In order to develop Cry1Ah protein certified reference material, it is urgent to establish a preparation and purification system. In this study, a system for preparing Cry1Ah protein by Bt expression system was optimized, and a high-purity Cry1Ah protein (size exclusion chromatography purity: 99.6%) was obtained by ion-exchange chromatography and size exclusion chromatography stepwise purification. The results of biological activity assay showed that there was no significant difference in the insecticidal activity of purified Cry1Ah protein and protoxin against diamondback moths (Plutella xylostella). Finally, the amino acid sequence of the activated Cry1Ah protein was determined using Edman degradation and mass spectrometry. In summary, the obtained Cry1Ah pure protein can be used for the development of protein reference materials.
Animals ; Bacillus thuringiensis ; Bacterial Proteins ; Cryptochromes ; metabolism ; Endotoxins ; Hemolysin Proteins ; Moths ; Pest Control, Biological ; Plants, Genetically Modified

Objective: To investigate the key factor(s) of hepatitis B virus X protein (HBx) promoting B7-H6 gene activation. Methods: The DNA fragments of the B7-H6 promoter were amplified from the human genomic DNA using polymerase chain reaction(PCR). Products of PCR were digested by KpnI and HindⅢ, and inserted into luciferase reporter vector (pGL3-Basic). The correctness of the recombinant plasmid pGL3-B7-H6 was confirmed by sequencing. Human hepatoma cell line HepG2 were co-transfected with pGL3-B7-H6 and the eukaryotic expression vectors (pCMV-HBs、pCMV-HBc and pCMV-HBx), and the relative luciferase activity was detected.The different dose of HBx expression plasmids were transfected into the HepG2 cells, and the expression of B7-H6 protein were determined by Western blot. Results: A period of 2.2 Kb of B7-H6 promoter region were successfully cloned into pGL3-Basic, which was confirmed by DNA sequencing. The relative luciferase activity was significantly higher in the cells transfected with pGL3-B7-H6 than that in the cells transfected with the empty vector pGL3-Basic (5.24±1.25 vs. 1.12±0.31, P=0.005). The relative luciferase activity in HepG2 cells co-transfected with pGL3-B7-H6 and pCMV-HBx was remarkably increased compared with the control group (17.60±3.36 vs. 6.73±1.36, P=0.001). Western blot further demonstrated that HBx protein can significantly enhance B7-H6 protein expression. Conclusions Hepatitis B Virus X proteins enhanced B7-H6 promoter activity and promoted B7-H6 protein expression.

Objective To evaluate the clinical effects of fiberoptic bronchoscopy and bronchoalveolar lavage in the treatment of refractory Mycoplasma pneumoniae pneumonia.Method The clinical data of 55 children with refractory Mycoplasma pneumoniae pneumonia were retrospectively analyzed during February 2015 to February 2016.Results Among those 55 children,30 cases who received the treatment of bronchoscopy and bronchoalveolar lavage were assigned to treatment group,and the other 25 cases were assigned to control group.17 children in treatment group and 13 children in control group had lung lamellar shadow.After treatment,the improvement rates were 100％ and 69.2％ in treatment group and control group,respectively.There was significant difference between two groups (P=0.026).The total effective rate in treatment group was 96.7％,which was significantly higher than that of control group (64.0％) (P=0.006).The length of hospital stay were significantly shorter and average treatment fee were significantly lower in treatment group than those in control group (P＜0.01).There were no severe adverse reactions in treatment group.Conclusion The effects of fiberoptic bronchoscopy and bronchoalveolar lavage with local application of drug were remarkable in the treatment of refractory mycoplasma pneumonia,and there were no severe adverse reactions.

Objective To investigate the effect of cogl410 on brain edema around lesion foci after traumatic brain edema (TBI) in mice and possible mechanism.Methods APOE-knockout TBI mice (n =130) were divided into cog1410 group and saline group according to the random number table,with 65 mice per group.Several time points (0,1,3,5,and 7 days) after TBI,13 mice were sacrificed in each group.TBI was induced with controlled cortex injury to the mice.A single injection of cog1410 solution or saline was administered via the caudal vein in 30 minutes after TBI.Levels of aquaporin-4 (AQP-4) around the lesion tissues were measured using the Western blot and q-PCR methods.Brain water content was determined by the dry-wet weight method.Results Brain water content in cog1410 group and saline group increased after TBI,reached the peak at day 3 [(81.184 ±0.009)％ vs (84.184 ± 0.014) ％] and normalized at day 7 [(76.018 ± 0.003) ％ vs (77.790 ± 0.012) ％] (P ＜ 0.05).There were almost no changes in AQP-4 mRNA expression in saline group after TBI.Whereas in cog1410 group,AQP-4 mRNA was greatly down-regulated at day 3 (0.278 ±0.014),increased greatly at days 5 and 7 after TBI (1.744 ± 0.014,1.782 ± 0.003) (P ＜ 0.05).Level of AQP-4 protein in saline group was increased at day 1 (0.491 ±0.060),reached the peak (0.605 ±0.099),and gradually returned to the preinjury level at days 5 and 7 (0.434 ± 0.042,0.358 ± 0.034).By contrast,level of AQP-4 protein in cog1410 group revealed no notable changes at day 1,slight increase at day 3,significant increase at day5 (1.079±0.090),and apeak at day7 (1.247±0.210) (P＜0.05).Conclusion cog1410 can significantly alleviate brain edema around the lesion foci of mice with TBI,as may be achieved by altering the mRNA and protein levels of AQP-4.

Objective To investigate the correlation of angiotensin Ⅱ type 1 receptor (AT1R) gene polymorphism with AT1R expression levels and brain edema after hypertensive intracerebral hemorrhage .Methods 45 operative patients with hypertensive intracerebral hemorrhage in the Affiliated Yongchuan Hospital of Chongqing Medical Univercity from December 2011 to August 2012 were collected as the experimental group and 45 operative patients with refractory epilepsy weres selected as the control group .The venous blood in the two groups were collected for detecting the AT 1R gene polymorphism ;The brain tissue was taken from lesions in operation ,then AT1R mRNA concentration was determined by RT-PCR and the AT1R protein level was determined by Western blot ;Head CT was performed on postoperative 1 ,3 ,5 d;the degree of cerebral edema was reflected by CT value . Results The levels of two kinds of genotype AT1R mRNA in the experimental group had no statistically significant difference(P>0 .05);the operative area CT value of AC genotype was significantly lower than that of AA genotype with statistical difference (P<0 .05);the ATIRmRNA of various genotypes ,protein level and cerebral edema in the control group had no statistical differences . Conclusion The AT 1R gene polymorphism has no obvious correlation with the concentration expression of AT 1R mRNA in the brain tis-sue;there is correlation between AT 1R protein level and AT 1R protein level and the cerebral edema degree in the brain tissue .

Objective To construct a eukaryotic expression plasmid of pcDNA 3.1-Ag85A-CD226, and to use it as DNA vaccine then further evaluate its immunogenicity through oral administration in a mouse model.Methods The CD226-PCR2.1-ToPo plasmid was used as the template to clone CD 226 gene by PCR.The CD226 gene was then inserted into pcDNA 3.1-Ag85A plasmid to construct the recombinant plas-mid of pcDNA3.1-Ag85A-CD226.After identified by restriction enzyme analysis and sequencing , the re-combinant plasmid was transfected into HEK 293 cells by using lipofection .The expression of Ag85A-CD226 gene in HEK293 cells was detected by RT-PCR, Western blot and indirect immunofluorescence assay .The purified recombinant plasmid was used to prepare the Ag 85A-CD226 DNA vaccine by liposomal encapsula-tion.The vaccine was administered intragastrically to mice .The activities of NK cells , the cytokine levels in the supernatants of spleen cell cultures and the mRNA level of cytokines in the intestines were evaluated to analyze the immunogenicity of Ag85A-CD226 DNA vaccine.Results The Ag85A-CD226 DNA vaccine was prepared successfully .The expression of Ag85A-CD226 fusion protein was detected in HEK293 cells.The activities of NK cells from mice vaccinated with Ag 85A-CD226 DNA vaccine were higher than those from other control groups (P<0.01).The level of TNF-α, IFN-γand IL-2 in the supernatants of spleen cell cul-tures and in the intestines were significantly up-regulated in comparison with other control groups ( P <0.01).The level of IL-4 in the supernatants of spleen cell cultures was down-regulated in the experimental group (P<0.01), but the level of IL-4 in intestines showed no significant difference among the five groups (P>0.05).Conclusion The Ag85A-CD226 DNA vaccine could significantly enhance Th1 type immune responses systemically and in the intestine as in comparison with those vaccinated with single dose of Ag 85A DNA vaccine or CD226 DNA vaccine.