OBJECTIVETo investigate the effects of adenovirus-mediated PTEN and P27 on the invasion of PC-3 in vitro and angiogenesis, along with their synergy in the treatment of prostate cancer.

METHODSRecombinant adenovirus vectors of the human tumor suppressor genes PTEN and P27 were constructed. The replication-incompetent recombinant adenovirus was packaged and propagated in HEK293 cells. The viral titer was examined by plaque assay and the mRNA and protein expressions of PTEN and P27 in human prostate cancer cell line PC-3 infected with Ad-PTEN and Ad-P27 were determined by RT-PCR and Western blot respectively. The invasion of PC-3 cells in vitro was examined by Boyden chamber assay. MTT assay was used to testify the effect of supernatant from PC-3 infected with Ad-PTEN and Ad-P27 on the proliferation of endothelial cells ECV-304 and the CAM test was used to testify the effect of PTEN and P27 on angiogenesis. The difference between the combined therapy group and the single gene therapy group was also examined.

RESULTSThe viral titers of Ad-PTEN and Ad-P27 were 1.8 x 10(7) pfu/ml and 1.2 x 10(9) pfu/ml respectively. Adenovirus infection verified that the mRNA and protein expression of PTEN and P27 were steady in human PC-3 cells. The invasion in vitro of PC-3 cells was significantly inhibited by infection with Ad-PTEN or/and Ad-P27. CAM and MTT assays of ECV-304 confirmed that the supernatant from PC-3 cells infected with Ad-PTEN or/and Ad-P27 could inhibit the angiogenesis effectively. There was a significant difference between the combined therapy group and the single gene therapy group.

CONCLUSIONThe combined gene therapy of Ad-PTEN and Ad-P27 plays a synergistic role in inhibiting the invasiveness of PC-3 cells and angiogenesis.

Adenoviridae ; genetics ; Cell Line, Tumor ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; Humans ; Male ; Neoplasm Invasiveness ; PTEN Phosphohydrolase ; genetics ; Prostatic Neoplasms ; blood supply ; pathology ; Transfection

Objective To investigate the phenotypes and genotypes of a hereditary protein C(PC) deficiency pedigree.Methods Imrnunoassay(ELISA)was used for PC antigen and PS antigen; Immunoturbidimetry assay was used for measuring AT antigen;Chromogenic substrate assay was used for measuring the activity of PC,PS and AT in Sysmex 1500 automatic Blood Coagulation Analyzer.Polymerase chain reaction(PCR)for amplification of the fragment of each exon and side sequences of PC gene in 10 members of the 3 generations;Direct DNA sequencing was used to examine the mutation site.Results Among 10 members of the 3 generation pedigree,8 of them had a PC:Ag level of 1.06-1.92 mg/L(normal references 3.00-6.00 rag/L),the activity of PC was between 41% and 67%(normal references 70%- 140%),which was significantly lower than the normal references while the levels of PS:Ag,PS:A,AT:Ag and AT:A were all within normal range.DNA sequencing analysis showed that there was a G to T mutation in exon IX of the PC gene at 12 918 position in 8 members.This mutation resulted in the substitution of terminator TGA for TGG which encoding tryptophan at 372 amino acid.There was a polymorphism in 2 405C/ T,2 418A/G,2 583A/T in the promotor area.Conclusions This pedigree is a type I hereditary protein C deficiency.There is a G12 918T mutation in exon IX of PC gene.This mutation is reported for the first time and there is a polymorphism in 2 405C/T,2 418A/G,2 583A/T in the promotor area.

OBJECTIVETo detect the mutations of Krit-1 gene that cause familial cerebral cavernous malformation (CCM) in the Han ethnic origin.

METHODSThe subjects were hospitalized in the Department of Neurosurgery, Tiantan Hospital affiliated to Capital University of Medical Sciences. Two families (A and B) and 8 apparently sporadic individuals affected with CCM were screened for mutations of Krit-1 gene. Members of the family CCM have a wide range in age of onset with seizures, headaches and skin lesions. The gene was screened by PCR amplification of 16 exons and mutation was detected by direct sequencing.

RESULTSIn family A samples, analysis of the Krit-1 gene revealed a new point mutation in exon 14 [a heterozygous C to G transition at nucleotide 1 289 (counting from the start codon or nt 2 308 counting from the first nt of the mRNA, aligned according to Gene Bank AF388384)] which predicts the substitution of a premature termination codon for Serine at codon 430 (S430X), belonging a nonsense point mutation. No mutation was identified in one of family A members as well as in any of the sporadic individuals with the exception of a single nucleotide polymorphism.

CONCLUSIONSReport the first family in the Han with CCM having a novel mutation in the CCM1 gene on the continent of Asia. The newly identified mutation creates a premature termination codon and is predicted to produce a truncated Krev1 interaction-trapped 1 protein, KRIT1. This result allows efficient presymptomatic molecular diagnosis.

Adult ; Base Sequence ; Child ; Child, Preschool ; Female ; Hemangioma, Cavernous, Central Nervous System ; genetics ; pathology ; Humans ; KRIT1 Protein ; Male ; Microtubule-Associated Proteins ; genetics ; Middle Aged ; Molecular Sequence Data ; Mutation ; Proto-Oncogene Proteins ; genetics

OBJECTIVETo explore the effect of the improved plastic and reconstruction of the anus in situ.

METHODSImproved plastic and reconstruction of anus in situ was performed in 38 cases of low rectal cancers operated while Miles radical operation. Improvement includes: (1) The internal sphincter was rebuilt with 4 layers of muscle layer of the endmost of colon. (2) The last of gracilis was divided into 2 parts to reconstruct the superficial part and deep part of external sphincter muscle. (3) The rectum cape improvement is to firmly stitch the levator ani outside the external sphincter muscle in front of the colon. (4) The rectum valve is improved into three artificial rectum valves.

RESULTSThe form and function and their long term survival rate were good, the rate of superior anus function was 94.73%.

CONCLUSIONIt mains the results of improved plastic and reconstruction of anus in situ is near that of normal persons.

Adult ; Aged ; Anal Canal ; surgery ; Female ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Rectum ; surgery

Adult ; Antiviral Agents ; administration & dosage ; therapeutic use ; Hepatitis C ; complications ; drug therapy ; Humans ; Kidney Transplantation ; methods ; Male ; Premedication ; Renal Dialysis ; Uremia ; complications

Objective:To construct a scientific research evaluation model through principal component analysis, and to explore scientific research evaluation methods for hospitals.Methods:The professional title, educational background, positions and scientific research output information of the scientific research personnel in the First Hospital of Jilin University from 2019 to 2020 were collected. Delphi expert consultation was used to determine the assignment value of each variable, and use SPSS 21.0 software was used to build the principal component analysis model and conduct model verification.Results:The study collected a total of 1 882 researchers′ information. The KMO value of the validity test and the Bartlett sphere test meet the requirements of principal component analysis (KMO=0.731, P<0.05); the model obtained a total of 7 principal components. Among them, principal component 1 represents researchers who published SCI papers, applying for national, provincial and ministerial level scientific research projects, and their part-time positions in academic societies. The second principal component represents the status of applying for patents and publications, and the third principal component represents the status of the awards. The scores of scientific research output of researchers were summarized and sorted according to disciplines, according to which the neurology, endocrinology and metabolism, neurosurgery, general surgery and orthopedics ranked better. The model verification results found that researchers with senior professional titles and doctoral degrees had the highest median weighted comprehensive score( P<0.05), suggesting that scholars with higher professional title levels and higher education received higher comprehensive scientific research output scores. Conclusions:The scientific research evaluation model constructed by this study can provide scientific data reference for the hospital scientific research evaluation.

Objective To identify prognostic factors ofmotorfunctionafter surgery of metastatic spinal cord compression (MSCC).Methods The clinical data of 681 patients with spinal metastases from January 2008 to December 2017 were retrospectively analyzed.According to inclusion and exclusion criteria,a total of 206 patients with spinal metastatic were included.Postoperative neurological function was assessed using Frankel classification.The influence of age,gender,preoperative status,number of spine metastases,location of spinal metastases,visceral metastases,bone metastases,primary tumor type,interval from symptom to surgery,time of developing motor deficits,interval from primary tumor diagnosis to MSCC,preoperativethe Eastern Cooperative Oncology Group performance status (ECOG-PS),Karnofsky Performance score (KPS) and surgical procedures on postoperative function outcomes were explored.Results 140 (68.0％) patients were able to walk postoperatively compared with 88 (42.7％) patients preoperatively.Moreover,in 89.8％ of all patients,79 ambulatory patients maintained ambulation after treatment.The univariate analysis according to Ordered-logit model showed thatnumber of spine metastases,location of spinal metastases,preoperative ECOG-PS,preoperative KPS,interval from symptom to surgery and time of developing motor deficits were related with posttreatment motor functions.The multivariable analysis showed that number of spine metastases (OR=2.03;95％CI:1.12-3.33;P=0.04),preoperative ECOG-PS (OR=4.84;95％CI:2.42-8.15;P=0.038),interval from symptom to surgery (OR=3.78;95％CI:3.12-9.15;P=0.024),time of developing motor deficits(OR=2.75;95％CI:1.22-3.89;P=0.01) were independent prognostic factors for function outcomes.Conclusion 1-2 levels of metastasis,Interval from symptom to treatment ≥ 48 h,time of developing motor deficits ≥7 d,and ECOG-PS 1-2 can be considered as the most significant positive prognosticfactors for post-treatment ambulatory status.Spinal metastasis should have a higher priority,and immediate intervention should be started before the development of irreversible neurologic deficits.Increasing awareness of early symptoms and earlier screeningwith regular outpatient review might make a difference for patients with MSCC.Consequently,the identified prognostic factors can be considered as apreoperative assessment tool to predict the neurologic outcomeand guide clinical treatment for individual patients with MSCC.

OBJECTIVETo investigate the relevant factors of liver histological changes in chronic hepatitis B (CHB) patients with mildly elevated ALT and to explore the clinical values of these factors on anti-viral treatment.

METHODSA total of 152 CHB patients with mildly elevated ALT (less than 2 x ULN) who underwent liver biopsy were included in the study. Correlations between routine laboratory markers, liver histological inflammation grade and fibrosis stage were statistically assessed by Spearman correlation analysis, one-way ANOVA, area under the curve (AUC) of the receiver operating characteristic curves (ROC) and Logistic regression statistical analysis.

RESULTSAll patients in the study showed various hepatic histological damages. Among the 152 patients 50 (32.9%) were found with inflammation grade 1 (G1), 42 (27.6%) with G2, 46 (30.3%) with G3 and 14 (9.2%) with G4. 16 patients (10.5%) were found with fibrosis stage 2 (S2), 25 (16.5%) with S3 and 41 (27.0%) with S4. Routine laboratory markers Alb, BPC and WBC were significantly correlated with hepatic histological inflammation grade and fibrosis stage. Marked liver fibrosis and moderate to severe liver damage were significantly higher in patients aged more than 40 years as compared to those less than 40 years of age (P = 0.002, P = 0.010). The regression equation P = 1/[1+e-(9.36250-1625Alb-0.0234BPC)] was established with sensitivity and specificity of 83.3% and 65.0%, respectively.

CONCLUSION67.8% of CHB patients with mildly elevated ALT have significant injury to the liver tissue. CHB patients aged more than 40 years have a significant increase of marked liver fibrosis and moderate to severe liver damage. The regression equation is valuable to predict whether CHB patients need antiviral therapy or not.

Adolescent ; Adult ; Alanine Transaminase ; metabolism ; Female ; Hepatitis B, Chronic ; enzymology ; metabolism ; pathology ; Humans ; Liver ; pathology ; Liver Cirrhosis ; pathology ; Male ; Middle Aged ; Young Adult

Hyperbaric oxygen therapy is a method of breathing pure oxygen or high-concentration oxygen in a highpressure environment to treat hypoxic diseases and related diseases. According to clinical verification, this therapy has an irreplaceable effect on certain diseases and has gradually become a comprehensive clinical treatment. One of the main methods of certain diseases is widely recognized by the medical field at home and abroad. The development history, treatment principles, key technologies, and future development trends of hyperbaric oxygen are discussed in detail, provide a research direction for the development of hyperbaric oxygen therapy in the future, and at the same time, it has also improved physicians' awareness of hyperbaric oxygen therapy, so as to improving Industry influence.
Hyperbaric Oxygenation ; Oxygen/therapeutic use* ; Research Design

Objective To explore the effect of scutellarin on lipopolysaccharide(LPS)induced neuroinflammation in BV-2 microglia cells.Methods BV-2 microglia were cultured and randomly divided into 6 groups:control group(Ctrl),cyclic GMP-AMP synthetase(cGAS)inhibitor RU320521 group(RU.521 group),LPS group,LPS+RU.521 group,LPS+scutellarin pretreatment group(LPS+S)and LPS+S+RU.521 group.The expressions of cGAS,stimulator of interferon gene(STING),nuclear factor kappa B(NF-κB),phosphorylated NF-κB(p-NF-κB),neuroinflammatory factors PYD domains-containing protein 3(NLRP3)and tumor necrosis factor α(TNF-α)in BV-2 microglia were detected by Western blotting and immunofluorescent double staining(n= 3).Results Western blotting and immunofluorescent double staining showed that compared with the control group,the expression of cGAS,STING,p-NF-κB,NLRP3 and TNF-α in BV-2 microglia increased significantly after LPS induction(P<0.05),while the expression of cGAS,STING,p-NF-κB,NLRP3 and TNF-α in LPS+S group were significantly lower than those in LPS group(P<0.05).Treatment with cGAS pathway inhibitor RU.521 showed similar effects as the pre-treatment group with scutellarin.In addition,the change of NF-κB in each group was not statistically significant(P>0.05).Conclusion Scutellarin inhibits the neuroinflammation mediated by BV-2 microglia cells,which may be related to cGAS-STING signaling pathway.