Objective To investigate the correlation between Dishevelled protein expression and the proliferation and invasion of glioma cells.Methods 67 cases of brain glioma specimens were collected.The expression of Dishevelled protein was detected with immunohistochemical method.The immunoreactivity score (IRS) of Dishevelled protein,and proliferation index (PⅠ) and invasion index (Ⅱ) were measured and their correlations were analyzed.Results The positive rate of Dishevelled protein in glioma was 65.7 ％ (44/67).IRS,PⅠ and Ⅱ were 4.15±3.13,(30.93±17.92) ％,(20.38±13.36) ％,respectively.Both PⅠ and Ⅱ significantly increased with an increase in the pathological grade of brain glioma (P ＜ 0.001).Furthermore,PⅠ and Ⅱ were significantly higher in the Dishevelled protein-positive group than those in the Dishevelled protein-negative group [(38.27±17.60) ％ vs (16.02±8.92) ％ of PⅠ and (30.03±13.81) ％ vs (10.63±4.41) ％ of Ⅱ,respectively,P ＜ 0.001].PⅠ and Ⅱ of glioma cells were positively correlated with IRS of Dishevelled protein (r =0.940 between PⅠ and IRS,and r =0.953 between Ⅱ and IRS,respectively).Conclusion Dishevelled protein plays an important role in the proliferation and invasion of brain malignant glioma.

Atopic dermatitis (AD) is a chronic, relapsing, inflammatory dermatosis with a variety of clinical manifestations and difficult to cure. Currently, many AD drug candidates have entered the research and development pipeline. In order to provide technical specifications for the clinical development of AD drugs, the Center for Drug Evaluation of National Medical Products Administration released the "Technical Guidelines for Clinical Trials of Drugs for AD Treatment" (Draft for Comments) in November 2022. Non-clinical pharmacodynamics evaluation is an important research before the drug enters clinical trials. Oxazolone (OXA)- and 2,4-dinitro-fluorobenzene (DNFB)-induced models are the most popular classical hapten-induced AD murine models, but variations of modeling are existing in the methods from different studies, including sensitization sites, haptens' dosages, the period of challenges, and the skin lesions severity evaluation as well. In this study, the investigation of OXA- and DNFB-induced AD murine models with various conditions of modeling was performed to compare the characteristics of hapten-induced AD murine models in the pathological process and severity according to the appearance of AD patients, and the guidance of pharmacodynamics evaluation of AD-therapeutic drugs in clinical trials as well, which may provide a proposal for AD treatment drug candidates in the non-clinical pharmacodynamics evaluation. All animal experiments were approved by the Animal Care & Welfare Committee of Institute of Materia Medica, Chinese Academy of Medical Sciences and Peking Union Medical College (approval No.: 00007782 and 00007784).

ObjectiveTo investigate the role of CD4+ CD25+ regulatory T cells(Treg) in peripheral blood and intestinal endotoxin (ET) in liver injury of rat severe acute pancreatitis (SAP).MethodsSixty male Wistar rats were randomly allocated into sham operation group(SO group) and severe acute pancreatitis group(SAP group).The rats in SAP group recevied the injection of Sodium Taurocholate(NaTc) into their far-end of bile-pancreatic duct and were sacrificed in 3-,6-and 12-hour,serum amylase(AMY) and alanine aminotransferase(ALT) determined by full-automatic biochemistry instrument,limulus reagent method is used to determine ET content in plasma,the proportion of Treg among peripheral lymphocytes was determined by Flow Cytometry(FCM),HE stain is used to observe pathological changes in liver and pancreas,the protein of Foxp3 activity was evaluated by immunohistochemistry staining,and the relationships between these indicators were analyzed using Pearson correlation analysis test.ResultsHistopathologic examination and the level of ALT revealed the occurrence of pancreatic inflammation and pathological changes of liver in SAP group.The percentage of Treg in SAP groups significantly increased at 3 h(2.26％ ±0.32％),6 h(2.36 ％ ±0.48％)and 12 h(2.80％ ±0.35％) comparing to the SO groups(P＜0.01) ; plasma ET levels compared with the SO group was statistically significant (P＜ 0,01 ),and 12 h (0.85 ± 0.11) compared to,3 h (0.74±0.11) and 6 h (0.78-±-0.07) was no significant difference (P＞0.05).The expression of Foxp3 protein on the livers were upregulated markedly.Pearson correlation analysis teat showed that quantities of Treg were positively proportional to the levels of ET(r=0.89,P＜0.01) after liver injury of SAP.ConclusionsSAP may lead to liver injury and the high plasma levels of ET and Treg in peripheral blood may play an important role in liver injury of SAP.

ObjectiveTo evaluate the effects of intranasal administration of recombinant interleukin-17A(rIL-17A) on the expressions of β-Defensin-2(Defb2) and macrophage inflammatory protein(MIP) in pneumococcal pneumonia murine models.MethodsTwenty-four BALB/c mice were divided randomly into normal control,pneumococcal pneumonia,and rIL-17A intervention groups ( n =8 ).Before intranasal (i.n) infection with Streptococcus pneumoniae,the mouse was treated with PBS or rIL-17Ai.n respectively.The mRNA levels of Defb2,MIP-1α and MIP-2β expression in lung tissue were detected by real-time quantity PCR.The numbers of bacteria and leukocytes in bronchoalveolar lavage fluid (BALF) were counted as well.And the concentrations of MIP-1α,MIP-2β,IFN-γ and IL-4 in BALF and in supematants of spleen cells and mediastinal lymph node cells were assayed by ELISA.Changes in lung tissue histopathology were observed with HE staining through light microscope.ResultsNeutrophil and macrophage numbers are higher in BALF of rIL-17A group,while the numbers of bacteria were lower,when compared with those in pneumonia group( P＜0.01 ).The expression of Defb2 and MIP-1α mRNA were up-regulated in lung after rIL17A treatment(P＜0.01 ).When compared with rIL-17A non-treated mice,rIL-17A treated mice secretedhigher levels of MIP-1α in lymph node cell culture supernatants( P＜0.01 ),higher levels of MIP-2β were observed in spleen cell and lymph node cell culture supernatants( P＜0.01 ),higher levels of IFN-T were detected in BALF( P ＜ 0.01 ) and culture supernatants of spleen cell ( P ＜ 0.01 ) and lymph node cell ( P ＜0.05),and higher levels of IL-4 were detected in BALF and spleen cell culture supernatant(P＜0.01 ).Comparative analysis have not detect a significant irflammatory cell increases in rIL-17A treated mice lung tissue; however the histopathological lesions were decreased.ConclusionIntranasal inoculation of rIL-17A can promote pulmonary neutrophil and macrophage recruitment and bacterium clearance,Intranasal inoculation of riL-17enhances the host defense against Streptococcus pneumoniae infection partly through increasing the expression levels of defensins,MIP,IFN-T and IL-4 etc.

Objective To investigate the indications, techniques and results of laparoscopic hepatectomy. Methods The clinical data and follow-up results of 463 patients who received laparoscopic hepatectomy at our institute were retrospectively analyzed. Results From March 1, 2007 to March 31, 2011, 463 cases of laparoscopic hepatectomy were successfully carried out. Of the 463 patients,165 were with primary liver cancer, 29 with metastatic liver cancer, 143 with hepatic hemangioma, 81with hepatolithiasis and 45 with other benign liver diseases (including hepatic angiomyolipoma, hepatocellular adenoma, focal nodular hyperplasia and chronic liver abscess). The surgical approaches included laparoscopic left lateral lobectomy (93 cases), left hepatectomy (71 cases), extended left hepatectomy (4 cases), right hepatectomy (29 cases), right posterior lobectomy (24 cases), hepatectomy of segment Ⅵ (56 cases), extended right hepatectomy (2 cases), central hepatectomy (8 cases) and hepatectomy of segments Ⅶ/Ⅷ, Ⅳa, caudate lobe and the junction of segment Ⅵ and Ⅶ (41 case).Nonanntomic and wedge resection were performed on 121 patients, and combined resection on 14 patients. The mean operation time, blood loss, length of hospital stay and incidence of postoperative complications were (244.71 ± 105. 07) minutes, (460. 26±425.81) ml, (15.51 ±4.36) days and 9.29%, respectively. And no operative death occurred. In the 194 cases with malignant liver lesions,185 cases were followed up for 2 to 50 months. The 1 year and 3 year overall and disease free survival rate were 90. 8% and 87.9% , 84.2% and 73. 7% respectively. Conclusions As a means of minimally invasive surgical approach, laparoscopic hepatectomy can be selectively adopted for the treatment of all kinds of liver diseases which located at different parts of the liver, with the advantages of smaller trauma, quick recovery and cosmetic benefits. The short-term results of laparoscopic hepatectomy is superior to and its long-term results is equal to that of open surgery. Benign liver diseases, small hepatocellular carcinoma and metastatic liver cancer are the good indications for laparoscopic hepatectomy.

Objective To express and purify mouse interleukin 17A(mIL-17A) in E. coli and to analyze its ability of stimulating macrophage inflammatory factors expression. Methods The coding gene of mouse mIL-17A mature protein was amplified from mouse spleen cells by RT-PCR. PCR product was cloned into the prokaryotic expressing vector pET28a, and the resulting recombinant plasmid pET28a/mIL-17a was then transformed into the host E. coli strain BL21(DE3) for expression. The mIL-17A protein was identified by SDS-PAGE and Western blot. The recombinant protein was purified by the Ni-NTA affinity chromatography, and was further tested on the stimulation of cytokine and chemokine of RAW264.7 cells by ELISA and real-time quantity PCR in vitro. Results The mIL-17A with bioactivity was over-expressed and purified successfully, and the results of real-time PCR and ELISA showed that recombinant mIL-17A stimulated macrophage mRNA upregulation of IL-6, defensin β2 and Cxcl3 and secretion of defensin β2, Ccl3, Cxcl3,IFN-γ, IL-6 and IL-4. Interestingly, these effects could be blocked by the addition of anti-IL-17A neutralizing antibody partly. After treatment with mIL-17, 74. 87-fold of defensin β2 mRNA expression was increased comparing with that of untreated cells( P ＜0.01 ), while blocking with anti-IL-17A antibody the increase was only 5.4-fold(P ＜ 0.01 ). Conclusion The recombinant mIL-17A has a strong stimulation on secretion of cytokine and chemokine of macrophage, that maybe result to the enhancement of anti-infection ability of macrophage.

Objective To validate the feasibility of the simulationmethod and the reliability of thesimulationresult through comparison between simulation and measurement of the energy spectrum from medical diagnostic X-ray (RQR-Radiation qualities in radiation beans emerging from the X-ray source assembly).Methods A simplified model of the medical diagnostic X-ray RQR radiation quality was established using code of BEAMnrc.The energy spectrum of the same RQR radiation quality were measured through a plane high-purity germanium spectrometer,and compared with the simulationresult.Results The difference of spectral distribution between measurement and simulation was less than 3％,in spite of the convolution processing not happened to the pulse height distribution measured by the spectrometer.And the spectral distribution,fluence,energy fluence,means energy distribution of the radiation was obtained using the code of BEAMDP.Conclusions As indicated above,it is possible to use the simulation of the energy distribution as a foundation for the establishment of X-ray RQR radiation quality.

Objective To explore the localization of epileptogenic focus and select the appropriate surgical procedures for post-traumatic epilepsy. Methods The clinical data of 21 patients with post-traumatic epilepsy were studied retrospectively. Epileptogenic focus was located by comprehensively analyzing data of electro-neurophysiology, neurological imaging and clinical manifestation. Surgical procedures were performed in all patients, including resection of lesion and peripheral cortex in 12 patients, epileptogenie focus resection plus low power bipolar coagulation in five, anterior temporal iobectomy plus amygdalohippocampectomy in three and corpus callosotomy in one. Results All patients were followed up from 6 months to 3 years, which showed satisfactory outcome in eight patients, marked improvement in six, improvement in five and slight improvement in two. The total effective rate was 90%. Conclusions Surgical procedure is important for intractable post-traumatic epilepsy. The good efficacy depends on precise localization of epileptogenic focus and combined application of various surgical procedures.

Objective To investigate the effects of the microbubbles combined with different mechanical index ultrasound irradiation on ultrastmcture and migration of colon cancer cells.Methods The experimental study was conducted.Colon cancer cells inn vitro (Lovo ceils) were cultured and divided into 4 groups,ceils in the A group were not treated and cells in the B,C and D groups were treated by microbubbles combined with different mechanical index ultrasound irradiation (mechanical index were 0.20,0.80 and 1.45).The changes of ultrastructure and migration of cells were observed using laser scanning confocal microscope and MilliceIl-PCF cell culturechamber method,respectively.Measurement data with normal distribution were represented as (x)±s.Comparisons among groups were analyzed by the one-way ANOVA.Pairwise comparisons were done by the t test.Results (1)Effects of the microbubbles combined with different mechanical index ultrasound irradiation on ultrastructure of Lovo cells:Lovo cells in the A group showed big nucleus,less plasma,regular arrangement,jagged-like or more irregular varicosity surrounding nucleus,twisted euchmmatin,expansion of nucleus cisternal space,homogeneous distribution and normal development of granular soil and clear mitochondrial ridge-like structures.Lovo cells in the B group showed big nucleus with regular arrangement,obvious nucleolus margination,endoplasmic reticulum dilatation,normal development of mitochondrion and clear mitochondrial ridge-like structures.Lovo cells in the C group showed broadening nucleus space,abnormal nucleus with karyopycnosis,chromatin condensation,a few remaining or obvious dilatation of rough endoplasmic reticulum,typically consisting of different fragments or bubbles,cytoplasmic vacuoles changes and decreasing mitochondrial ridge-like structures.Lovo cells in the D group showed big and irregular nucleus,nucleolus margination,obvious endoplasmic reticulum dilatation,fewer mitochondrion with extended cell area and swelling shape,rare mitochondrial ridge-like structures with disordered or broken arrangement,even disappearing.(2) Effects of the microbubbles combined with different mechanical index ultrasound irradiation on migration of Lovo cells:Millicell-PCF cell culture chamber method showed that number of migration of Lovo cells were respectively 63±7,61±4,21±3 and 19±5 in the A,B,C and D groups,with a statistically significant difference (F=55.040,P＜0.05).There were no statistically significant difference in migration of Lovo cells between group A and B (t =1.571,P＞0.05) and between group C and D (t =2.013,P＞0.05).There were statistically significant differences in migration of Lovo cells between group A and C or D (t=7.861,10.652,P＜0.05) and between group B and group C or D (t=7.161,10.453,P＜0.05).Conclusion Microbubbles combined with high mechanical index ultrasound irradiation can make the ultrastructural alterations in the cancer cells,resulting in tumor cell degeneration and death,ultimately inhibit tumor cell migration and metastasis.

Aim To observe the effects of tetrahydro-biopterin ( BH4 ) on nitric oxide ( NO ) production in the kidney of type 2 diabetic nephropathy ( DN) mice, and to find a new target for the treatment of type 2 DN. Methods The 12 week-old db/db mice developed in-to DN phase were divided into 2 groups:DAHP group, subjected to intraperitoneal injection of 150 mg·kg-1 DAHP (n=8);DN group, subjected to intraperitone-al injection of same dose of normal saline containing 5% DMSO ( n = 6 ) . The age-matched db/m mice ( NS group) were subjected to intraperitoneal injection of same dose of normal saline containing 5% DMSO ( n =6 ) . Three groups of mice were treated for 7 days. Then the fasting blood-glucose, serum creatinine, u-rine protein and activity of iNOS were determined by chemical colorimetry. And the iNOS protein in renal cortex was determined by immunohistochemisty and western blot, respectively. BH4 was measured by HPLC method. NO level was determined by Griess method. Results The levels of fasting blood-glucose, serum creatinine, 24h urine volume, 24h urine pro-tein, BH4 , iNOS and NO in DN group were signifi-cantly higher than those in NS group;The levels of ser-um creatinine, urine volume, urine protein, BH4 , iN-OS and NO in DAHP group were significantly lower than those in DN group. Conclusion In the kidney of type 2 DN mice, the increased BH4 contributes to over-production of NO by the increased iNOS expression, and resultes in the increase of urine volume and urine protein.