OBJECTIVETo investigate the feasibility of promoting nerve regeneration by using microcapsulated NGF-expressing cells transplantation.

METHODSThe plasmid pcDNA3. 1 + /NGF, containing rat NGF gene, was transfected into the NIH 3T3 cell by using FuGENE6 transfection reagent. The genetically modified cells expressing NGF gene were enclosed within the alginate-polylysine-alginate (APA) microcapsules and then cultured in vitro. The growth and NGF secretion of the cells were measured periodically. At the same time, these microcapsulated NGF-expressing cells were transplanted into the injured sciatic nerve. The regeneration and functional recovery of the nerve were evaluated in 4 weeks, 8 weeks and 12 weeks after the operation.

RESULTSThe microcapsulated cells had survived and secreted the NGF in three months in vitro. In the group with microcapsulated NGF-expressing cells, the number of the regenerated axons was in large and the nerve fibers were arranged regularly. Compared to other groups, there was less scar , edema and monocytes found at the stoma in the goup. The moter nerve conductive velocity, nerve muscle-action potential and SFI were improved.

CONCLUSIONSThe microcapsulated NGF-expressing cells could significantly enhance the nerve regeneration and reduce inflammatory response of xenograft.

Animals ; Cell Transplantation ; Female ; Male ; Mice ; NIH 3T3 Cells ; Nerve Growth Factor ; biosynthesis ; genetics ; Nerve Regeneration ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVETo investigate the expression of telomerase in various lesions of adrenal cortex.

METHODSBy autoradiography-based telomeric repeat amplification protocol, telomerase expression was detected in 36 samples of adrenocortical lesions, including 29 cases adrenocortical adenoma (8 Cushing's syndrome, 17 aldosteronism and 4 nonfunctional adenomas), 5 cases of hyperplasia of adrenal cortex (presented with Chushing' syndrome), 2 cases adrenocortical carcinoma, and 4 samples of normal adrenal cortex.

RESULTSOf the 40 samples, 2 cases of adrenocortical carcinomas had telomerase expression, and the others had no telomerase expression detected.

CONCLUSIONSNo significant telomerase expression was found among different endocrine functional benign adrenocortical lesions. Telomerase expression may be used as an important marker of malignant adrenocortical tumor.

Adrenal Cortex ; enzymology ; Adrenal Cortex Neoplasms ; enzymology ; Adrenocortical Adenoma ; enzymology ; Biomarkers, Tumor ; analysis ; Cushing Syndrome ; enzymology ; Humans ; Telomerase ; analysis ; biosynthesis ; genetics

OBJECTIVETo analyze and optimize the gene rearrangement primers of different frame regions (FR) of immunoglobulin heavy chain (IgH) genes by bioinformatic methods and explore the application of these primers in the detection of paraffin-embedded lymphoma tissues.

METHODSThree pairs of primers from IgH FR1, FR2 and FR3 regions (P1c, P2A and P31, respectively) were selected as the B cell gene rearrangement primers after comparison of the gene fragments in 44 IgH variable and 6 joining regions. Using one pair of T cell receptor (TCR) gamma primer as the T cell gene rearrangement primer, 101 histopathologically confirmed lymphoproliferative samples including 80 B cell lymphomas, 14 T cell lymphomas, and 7 reactive proliferative lymph nodes were examined by PCR for gene arrangement. The DNAs from DG75 and Jurkat cell lines were used as the positive controls for B and T cell lymphoma, respectively, with those from reactive proliferative lymph nodes as the negative control.

RESULTSThe positivity rates of IgH primers (P1c, P2A and P31) in the 80 B cell lymphomas were 37.5% (30/80), 52.5% (42/80) and 70.0% (56/80), respectively, and only one of the 14 T cell lymphoma cases was positive for the primers, suggesting significant differences in the detection rates of B cell lymphomas by the 3 primers. The detection rate was increased to 83.9% by combining the results by P31 and P2A primers. No positivity was found in the proliferative reaction tissues.

CONCLUSIONPrimers from IgH FR3 region genes are more sensitive than that from the FR1 and FR2 regions in the detection of gene rearrangement in paraffin-embedded lymphoma tissues. The detection rates can be increased by combining the results with the primers for IgH FR3 with that of FR2.

DNA Primers ; Gene Rearrangement, B-Lymphocyte, Heavy Chain ; genetics ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Lymphoma, B-Cell ; diagnosis ; genetics ; pathology ; Lymphoma, Non-Hodgkin ; diagnosis ; genetics ; pathology ; Lymphoma, T-Cell ; diagnosis ; genetics ; pathology ; Male ; Paraffin Embedding

Objective To develop appropriate evaluation methods of local basic public health services which are suitable to county level and above.Methods Data on basic public health services of 1 1 cities in Zhejiang province in 201 2 was evaluated by different evaluation methods including weighted synthetic scored method,weighted synthetic index method, Weighted Technique for Order Preference by Similarity to Ideal Solution (Topsis ) and Weighted Rank -Sum Ratio (RSR).The consistency of evaluation results were tested by Kendall's coefficient of concordance W test.Combination evaluation was conducted to evaluate four single synthetic evaluation results through average method,weighted average combination evaluation method and hierarchical clustering analysis.Results Different synthetic evaluation methods had different evaluation results.However,in the order,the top two were all Hangzhou and Ningbo.Kendall's W test showed good consistence of four evaluation results.Rank of 1 1 cities were Hangzhou,Ningbo,Shaoxing,Jiaxing,Huzhou, Taizhou,Jinhua,Zhoushan,Lishui,Wenzhou and Quzhou based on combination evaluation value by average method, which was the same to the rank based on weighted average combination evaluation result.Eleven cities could be classified into four categories by hierarchical clustering analysis with statistical significance (P <0.01 ):Excellent (Hangzhou, Ningbo),Good (Huzhou,Jiaxing,Shaoxing),Middle (Zhoushan,Jinhua,Taizhou)and Poor (Wenzhou,Quzhou, Lishui).Conclusion These four synthetic evaluation methods used in this study are all suitable to county level and above in basic public health services evaluation.Various synthetic evaluation methods could be used in practice with combination evaluation of various evaluation results.Average method which is convenient and accurate is preferred when consistency of various synthetic evaluation results was testified.Hierarchical clustering analysis could be used for combination evaluation when no precise rank is needed.

Objective To investigate the mitochondrial damage and its effect in early stage of pressure ulcer in rats.Methods Forty rats were randomly divided into 5 groups(n=8), control group(Con group) rats without stress, the experimental group was treated with of 170 mmHg for 2 h and relax 0.5 h as one cycle(1C), experi-mental group was divided into 3C, 6C, 9C and 12C group.The pathological changes of the compressed muscle tissue of the rats in each group were observed by HE staining , Western blot was used to detect the expression of Bcl-2 and Bax in the compressed tissue , and the ultrastructure of muscle fibers and mitochondria were observed by transmission electron microscope .Results There were pathological damage and gradually increased in the ex-perimental groups, with the increase of compression cycle; the expression of Bcl-2 in each experimental group was significantly increased as compared with the control group(P<0.05), in the 3C group reached the peak, and then decreased; the expression of Bax was increased gradually with the increase of compression cycle ( P<0.05) , and in the 12C group reached the peak;with the increase of the compression cycle the muscle fibers of each experimental group appeared gradually increased pathological damage:disorder, dissolution and fracture, the ridge of the mitochondria disappeared, vacuolar degeneration, et al.Conclusions In the early stage of pres-sure ulcer in a rat , it brings occurred mitochondrial damage and induces apoptosis .

OBJECTIVETo examine the expression of nestin and neurogenin 3 (Ngn3), the markers of pancreatic stem cells, in the human fetal pancreas.

METHODSThe human fetal pancreas tissue of 12 and 14 weeks were examined for the expression of nestin and Ngn3 using the techniques of immunofluorescence dye and RT-PCR.

RESULTSBoth nestin and Ngn3 expressed widely in 12 and 14 weeks before in human fetal pancreatic tissue. In these positive cells there was no co-expressing insulin or glucagon. There were nestin and Ngn3 co-expressing cells in ducts but not in the islets. The results of RT-PCR also indicated the expression of nestin and Ngn3.

CONCLUSIONSThere was no expression of the markers of mature endocrine cells in the nestin and Ngn3 positive cells, and they were the marks of no-differentiation cells in the human fetal pancreatic tissue.

Basic Helix-Loop-Helix Transcription Factors ; biosynthesis ; genetics ; Fluoroimmunoassay ; Humans ; In Vitro Techniques ; Intermediate Filament Proteins ; biosynthesis ; genetics ; Microscopy, Fluorescence ; Nerve Tissue Proteins ; biosynthesis ; genetics ; Nestin ; Pancreas ; cytology ; embryology ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the genomic variation of Epstein-Barr virus (EBV) and its significance in nasopharyngeal carcinogenesis.

METHODSForty nasopharyngeal carcinoma (NPC) biopsy tissues were used for detection of EBV BamHI f variant and LMP1 XhoI-loss by polymerase chain reaction (PCR), nested PCR, and RFLP (restriction fragment length polymorphism). Forty-eight samples of peripheral blood mononuclear cells (PBMC) taken from apparently healthy adult individuals were used for detection of LMP1 XhoI-loss. Three samples of amplified LMP1 exon 1 DNA from B95-8 cell line and 2 NPC tissues (one having XhoI-loss and the other having Wt-XhoI/XhoI-loss) were sequenced.

RESULTSThirty out of the 40 NPC cases (30/40, 75%) harbored EBV BamHI f variant and the remaining 10 (10/40, 25%) harbored BamHI F prototype. Thirty out of the 39 NPCs (30/39, 76.9%) showed single EBV LMP1 XhoI-loss, 7 (7/39, 18.0%) showed single LMP1 Wt-XhoI (presence of a XhoI site in exon 1 of LMP1 gene, as in B95-8 cell line), and 2 (2/39, 5.1%) showed both LMP1 Wt-XhoI and XhoI-loss. Thirty-eight of the 39 NPCs (97.4%) showed EBV LMP1 XhoI-loss or/and BamHI F variant. In the NPC tissue (1 case only) showing the prototype of Wt-XhoI/BamHI "f", there were several base substitutions, including 5 missense mutations and 2 silent mutations present in LMP1 exon 3, on DNA sequencing. On the other hand, 10 out of the 48 samples of PBMC taken from apparently healthy individuals could be amplified successfully by nested PCR for detection of LMP1 XhoI site. All of these 10 samples carried the prototype of EBV LMP1 Wt-XhoI.

CONCLUSIONSThe majority of EBV present in neoplastic cells of NPC is of BamHI "f" variant and/or possesses LMP1 XhoI-loss, as compared with that in healthy individuals. This genomic variation of EBV may bear some roles in the development and progression of NPC.

Adult ; Aged ; Binding Sites ; genetics ; DNA, Viral ; genetics ; metabolism ; Deoxyribonuclease BamHI ; metabolism ; Deoxyribonucleases, Type II Site-Specific ; metabolism ; Female ; Herpesvirus 4, Human ; genetics ; isolation & purification ; Humans ; Male ; Middle Aged ; Mutation ; Nasopharyngeal Neoplasms ; virology ; Sequence Deletion ; Viral Matrix Proteins ; genetics

Objective:To analyze the epidemiological characteristics and genotypes of human rhinovirus (HRV) in patients with upper respiratory tract infection in Qingdao in the winter of 2020.Methods:Throat swab samples were collected from 101 patients with upper respiratory tract infection in Qingdao from November 2020 to January 2021. Quantitative PCR was used to detect 15 common respiratory viruses in the samples. HRV-positive samples were further analyzed with RT-PCR to amplify and sequence HRV VP4/VP2 gene. A phylogenetic tree was constructed based on the sequencing results and homology analysis was conducted.Results:Six common respiratory viruses were detected in the 101 patients. Thirty-four cases (34/101, 33.66%) were single pathogen infection and two cases were multiple infection (2/101, 1.98%). The positive rate of HRV was the highest (21.78%, 22/101). Twenty HRV VP4/VP2 sequences were successfully amplified. Phylogenetic analysis showed that there were 16 strains of HRV-A subtype and four strains of HRV-C subtype and 14 serotypes were involved.Conclusions:HRV was one of the leading viral pathogens causing upper respiratory tract infection in Qingdao in the winter of 2020 and the predominant subtype was HRV-A.

OBJECTIVETo investigate the change of the mortality of AMI and influence factors within 20 years.

METHODSClinic data of 134 AMI patients from 1980 to 1983, 354 AMI patients from 1990 to 1993 and 817 AMI patients from 2000 to 2003 were comparably analyzed.

RESULTSIn hospital mortality of AMI was 22.4% from 1980 to 1983, 14.4% from 1990 to 1993 and 9.2% from 2000 to 2003, respectively (P < 0.01). The decrease of in-hospital mortality in male was more significant than in female (P < 0.01). The corresponding factors for decrease of mortality were younger than 60 years old, first onset of AMI, successful rescue of cardiac arrest and reperfusion management of infarction relative artery. The disadvantage factor was female.

CONCLUSIONSImprovement of medical and reperfusion management of AMI conduced in significant decreases of hospital mortality.

Adrenergic beta-Antagonists ; therapeutic use ; Aged ; Angiotensin II Type 1 Receptor Blockers ; therapeutic use ; Angiotensin-Converting Enzyme Inhibitors ; therapeutic use ; Cause of Death ; Female ; Hospital Mortality ; Humans ; Inpatients ; Logistic Models ; Male ; Middle Aged ; Myocardial Infarction ; diagnosis ; mortality ; therapy ; Myocardial Reperfusion ; Prognosis ; Retrospective Studies ; Risk Factors

OBJECTIVETo establish DNA microarrays-based microRNA (miRNA) expression profiles of squamous cell carcinoma of larynx, using archived formalin-fixed paraffin-embedded tissue blocks, and to screen out and identify the differentially expressed miRNAs associated with the biological characteristics of this malignant disease.

METHODSTotal RNA was prepared from the formalin-fixed paraffin-embedded tissue blocks. After quality identification and fluorescent labeling, the RNA samples were hybridized with the Agilent human miRNA microarrays which contains 723 probes for human miRNAs. The data was processed with the softwares GeneSpring GX and R-Project.

RESULTSFrom the formalin-fixed paraffin-embedded tumor blocks collected, 24 RNA samples were obtained with the quality accorded to the requirement of miRNA microarray analysis, and both the hybridization and consequent data processing were accomplished. A total of 319 miRNAs were identified and among them 96 were detected in all the 24 formalin-fixed paraffin-embedded blocks of laryngeal carcinoma; and 5 differentially expressed miRNAs (false discovery rate < 0.05) were found to be associated significantly with the lymphatic metastasis of laryngeal squamous cell carcinoma (P < 0.05), including miR-23a(*), miR-28-5p, miR-15a, miR-16 and miR-425.

CONCLUSIONSHistopathological archives of well-annotated formalin-fixed paraffin-embedded tissue specimens are the valuable resources for miRNA study including to collect RNA samples for miRNA microarray analysis. A panel of differentially expressed miRNAs (miR-23a(*), miR-28-5p, miR-15a, miR-16 and miR-425) derived from the miRNA expression profile may serve as the potential molecular biomarkers for the prediction of metastasis development in laryngeal squamous cell carcinoma.

Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Gene Expression Profiling ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; pathology ; Lymphatic Metastasis ; MicroRNAs ; metabolism ; Oligonucleotide Array Sequence Analysis ; methods ; Paraffin Embedding