BACKGROUND: Studies have demonstrated that exogenous basic fibroblast growth factor (bFGF) has intensive effects to promote proliferation of gingival fibroblasts (GFs) cultured in vitro and the heeling of gingival wounds. OBJECTIVE: To investigate the effects of bFGF gene transfection on the biological performance of Beagle canine GFs. DESIGN, TIME AND SETTING: An observation and comparison in vitro experiment regarding cells was accomplished in Centre of Cell Biology and Development of Fujian Medical University and Department of Comparative Medicine in Fuzhou General Hospital of Nanjing Military Area Command of Chinese PLA from April to September of 2008. MATERIALS: Four beagle dogs, male, 12 months old, weighing 10-13 kg were used in this experiment, plRES2-EGFP-bFGF plasmid containing full-length human bFGF gene cDNA was constructed and conserved by our institution. METHODS: Free gingiva of the 2nd, 3rd and 4th premolars were excised from left upper jaw of Beagle dogs, dnsed with aseptic phosphate buffer four times, then cut into pieces and digested with 2.5 g/L pancreetin for 2 hours at 37 ℃. After the cantrifugation and supernatant removal, DMEM containing 10% fetal bovine serum was added to incubate on 6-well plate with coverlips in 5% CO2 incubator at 37 ℃. Logadthrnically growing cells were digested and passaged. GFs were transfected with pIRES2-EGFP-bFGF plasmid using liposome mediated method, while vacant plasmid transfection and un-transfection group served as controls.MAIN OUTCOME MEASURES: Proliferation and apoptosis feature of the GFs were evaluated by M'rE and AOEB, respectively. The activity of alkaline phosphatase was assayed by chemical coledmetry. RESULTS: All of three groups cells entered log phrase on three days after transfection. MTT results showed that the proliferation of GFs transfected with bFGF was greater than cells transfected with vacant vector and untransfected cells (P < 0.05). AO/EB dyeing showed the apoptosis rate of GFs transfected with bFGF was reduced compared with other two groups (P < 0.05). After bGFG gene transfection, the ALP activity remained unchanged and there was no significant difference compared with untransfected cells.CONCLUSION: The transfection of bFGF gene to GFs can promote the proliferation of GFs and depress the apoptosis. No promotion is present with regard to the GFs differentiation.

Objective To study the expression level and clinical significance of bcl-XL gene in childhood medulloblastoma.Methods The expression of Bcl-XL protein and bcl-XL mRNA were determined by immunohistochemical staining and in situ hybridization in 41 samples of medulloblastoma tissues,as well as 20 normal brain tissues.Results The positive rate of Bcl-XL protein(90.2%) and bclXL mRNA(95.1%) in medulloblastoma group were significantly higher than those in normal human brain tissues(all P

Age Factors ; Endometrial Hyperplasia ; pathology ; Endometrial Neoplasms ; pathology ; Endometrium ; cytology ; pathology ; Female ; Humans ; Menstrual Cycle ; Papanicolaou Test ; Postmenopause ; Vaginal Smears

Objective To investigate the expression and significance of tumor necorisis factor related apoptosis induled ligand receptor(TRAILR) in human craniopharyngioma.Methods The expression of TRAILR was determined by immunohistochemistry and in situ hybridization in 24 samples of craniopharyngioma and 16 samples of normal brain tissue.Results With low decoy receptor(DcR) expression in partial craniopharyngioma cells and low death receptor(DR) expression in partial normal brain cells,DR was expressed highly in all craniopharyngioma samples while DcR in most normal brain tissue. High DR expression and low DcR expression in craniopharyngioma tissue differed from low DR expression and high DcR expression in normal brain tissue(P

Objective To investigate histopathological changes and expression of integrin ?1 of sternomastoid muscle,and probe the mechanism and significance during disease process in congenital muscular torticollis(CMT).Methods Histopathological changes of sternomastoid muscle section stained with HE and Gomori silver staining were observed and the expression of integrein ?1 with immunohistochemistry was detected,and the expressive quantity and distribution with image analysis system was quantitive analyzed.Results 1.With light microscopy observation,the results showed that the fibrous degeneration of sternomastoid muscle could be summed up 2 kinds: A category displayed the myocytes atrophyed,and there were lots of connective tissue hyperplasy around myocytes,and the direction of fibrous arrangement was disordered,meanwhile there were lots of vessels and nervers hyperplasy,and eventually the myocytes shrank back and disappeared.B category displayed that the structure of cross striation or sarcomere disappeared or changed,and myocytes could maintained the outline and the sarcolemma were integrated,and then fibrous pathological changes of myocyte took place,and there were lots of fibroblast-like that had much more enations between fiber bundles.With Gomori silver staining,the major changes of fibrotic sternomastoid muscle showed that there were lots of collagenous fibers hyperplasy.The arrangements of collagenous fibers were disordered in A category and were well-arranged in B ca-tegory.2.With immunohistochemistry,the results showed the expression of integrin ?1 was weak positive in normal control group(125.7?5.167).In diseased groups,the results showed 3 different extents:the expression of integrin ?1 displayed stronger positive in A category myocytes(30.15?6.543),and the level of expression was significantly different from normal controls(P0.05).Conclusions The fibrous pathological changes of sternomastoid muscle are a complicated and gradually process,which may has different mode,and ingetrin ?1 may participated the process of pathological changes.

ObjectiveTo evaluate the efficacy of rehabilitation associated with local intramuscular injection of botulinum toxic A (BTX-A) on spastic cerebral palsy (CP).Methods60 children with spastic CP were divided into experimental group and control group with 30 cases in each group. Cases of experimental group were treated with rehabilitation associated with local intramuscular injection of BTX-A. Cases of control group were treated only with rehabilitation treatment. The therapeutic efficacies of two groups were evaluated with physician rating scale (PRS) and activities of daily living (ADL) evaluation systems.ResultsImprovement of clinical evaluations index-PRS and ADL in experimental group was much more significant than that of control group (P<0.05). ConclusionRehabilitation associated with local intramuscular injection of BTX-A can improve the efficacy of spastic CP.

It is often necessary to construct more than one recombinant plasmids when investigating the characteristics, physchemical features and functional mechanisms of genes or proteins. Repeated sub-cloning procedures including design of primers, enzyme digestion, ligation and verification of recombinant plasmids, have to be involved with. For this reason, it has become a tendency to developing new genetic vectors which can be used in multitude of experiments. Therefore, by using pIRES vector as a backbone, here we reported the construction of a mammalian expression vector: pCMV-Myc-IRES-EGFP which contains the N-terminal c-Myc epitope tag and the enhanced green fluorescent protein (EGFP) translated in an IRES-dependent manner. This novel vector can be used to testify the efficiency of cell transfection, to collect successfully transfected cell population via cytometry, to conduct transcription and translation in vitro, to purify target proteins or to trap their interactional proteins. The availability of this vector can facilitate function study of genes.
Apoptosis Regulatory Proteins ; genetics ; metabolism ; Base Sequence ; Blotting, Western ; Cell Line ; Cloning, Molecular ; Gene Expression ; Genes, myc ; genetics ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; metabolism ; Humans ; Microscopy, Fluorescence ; Molecular Sequence Data ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

With the emergence of drug resistant tuberculosis, it is very urgent to find novel anti-tuberculosis drugs, especially novel anti-drug-resistant tuberculosis drugs. Because of the slow growth and the need to work in a biosafty environment of Mycobacterium tuberculosis, the development of evaluation of drug effect is severely impeded. In order to solve these issues, non-pathogenic fast-growing Mycobacterium smegmatis is introduced as test organism. The inhA is one of a target of isoniazid (INH) overexpression or mutation of this gene in Mycobacterium tuberculosis conferring resistant to INH. A recombinant plasmid bearing inhA was constructed and electroporated into Mycobacterium smegmatis, using shuttle expression vector pMV261. Transformants were induced to express a protein of inhA, identified by SDS-PAGE. Results show that Mycobacterium smegmatis containing inhA plasmids exhibited 100-fold or greater increased resistance to INH, but it conferred no increased resistance to others first-line anti-tuberculosis drugs. Resazurin microtiter assay plate testing of Mycobacterium smegmatis susceptibility to drugs is a rapid, simple, and inexpensive method and could decrease color background of drugs by detecting fluorescence. It will be benefit for high-throughout screening of drugs of anti-isoniazid-resistant Mycobacteria.

Objective To evaluate the post-operative glucose level and insulin dose of patients undergoing total pancreatectomy.Methods From September 1980 to September 2014, 21 patients underwent total pancrea-tectomy in Peking Union Medical College Hospital, who were enrolled in our study.We reviewed the changes in their insulin dosage and glucose levels after operation, also summarized type and dose of insulin as well as glucose level in stable period.Results The required insulin dose reached peak within 4 days after surgery ( maximum dose 300 U/d).The average dose was (143.5 ±62.8) U/d and decreased gradually.During the perioperative period (needing parenteral nutrition), the blood glucose level fluctuated markedly (1.52-29.06 mmol/L) and the average level was (11.18 ±0.95) mmol/L.During the stable period ( without parenteral nutrition) , patients on average had (5.3 ±2.0) U of preprandial rapid-acting insulin and (8.1 ±2.9) U of long-acting insulin be-fore sleeping;the average fasting blood glucose was (6.69 ±1.48) mmol/L, 2 h postprandial blood glucose was (9.08 ±2.84) mmol/L, bedtime blood glucose was (9.66 ±2.49) mmol/L, and blood glucose level at night was (8.15 ±2.78) mmol/L.67%of the patients had 13 hypoglycemic episodes monthly on average.For those five followed-up patients, the average hemoglobin A1c was (6.15 ±1.20)%.Conclusions Patients undergoing total pancreatectomy may experience marked fluctuation of blood glucose level and short-term increase of insulin need which gradually decreases afterwards.After entering the stable period, the glucose level could be well-con-trolled but with frequent hypoglycemia.There is no diabetic ketoacidosis.

OBJECTIVETo explore the mechanisms of Qianjing Decoction in the treatment of oligoasthenospermia (OAS).

METHODSWe randomly divided 100 SPF male rats into five groups of equal number: normal, model, Huangjingzanyu, levocarnitine, and Qiangjing. OAS models were established in the animals followed by intragastrical administration of normal saline, ornidazole, Huangjingzanyu Capsules (200 mg per kg body weight per day), levocarnitine (100 mg per kg body weight per day), and Qianjing Decoction (10 g per kg body weight per day), respectively, qd, for 4 successive weeks. Then, we detected the concentration and motility of the epididymal sperm, obtained the contents of superoxide dismutase (SOD), malonaldehyde (MDA), glutathione peroxidase (GSH-Px), lactate dehydrogenase (LDH), α-glucosidase, and fructose in the epididymis, and determined the mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2) and succinate dehydrogenase (SDH) in the epididymal tissue of the rats by real-time PCR.

RESULTSThe concentration and motility of the epididymal sperm in the model, Huangjingzanyu, levocarnitine, and Qianging groups were (35.34 ± 4.22) x 10(6)/ml and (40.04 ± 7.05)%, (48.12 ± 5.56) x 10(6)/ml and (62.46 ± 7.12)%, (47.14 ± 4.87) x 10(6)/ml and (63.23 ± 6.34)%, and (50.25 ± 5.08) x 10(6)/ml and (66.34 ± 7.58)%, respectively, all significantly lower than in the normal group ([53.05 ± 4.55] x 10(6)/ml and [70.20 ± 8.54]%) (P < 0.05), but remarkably higher in the Huangjingzanyu, levocarnitine, and Qiangjing groups than in the model rats (P < 0.05). Compared with the thinned epididymal lumen walls, decreased sperm count, and disorderly and loose arrangement of the lumens in the OAS models, the rats in the Huangjingzanyu, levocarnitine, and Qiangjing groups showed evidently thicker epididymal lumen walls, with the lumens full of sperm cells and arranged regularly and compactly, similar to those of the normal rats. The levels of SOD and GSH-Px were significantly lower but that of MDA markedly higher in the model rats ([84.12 ± 23.25], [10.56 ± 3.02], and [14.04 ± 2.06] nmol/mg) than in the normal group ([110.04 ± 19.56], [17.25 ± 3.56], and [8.87 ± 1.35] nmol/mg) (P < 0.05), while the former two indexes remarkably higher and the latter one significantly lower in the animals treated with Qiangjing Decoction ([120.56 ± 23.68], [16.34 ± 3.12], and [8.45 ± 1.56] nmol/mg), Huangjingzanyu Capsules ([115.34 ± 21.35], [15.23 ± 3.67], and [8.33 ± 1.54] nmol/mg), and levocarnitine ([116.67 ± 22.67], [15.35 ± 3.45], and [8.05 ± 1.78] nmol/mg) than in the models (P < 0.05). The levels of fructose, LDH and α-glucosidase were decreased markedly in the OAS models ([100.22 ± 12.12] mg/[ ml x g], [322 ± 46.13] U/[ ml x g], and [10.48 ± 2.33] U/[ml x g]) as compared with the normal rats ([128.12 ± 13.45] mg/[ml x g], [428 ± 35.12] U/[ml x g], and [15.34 ± 3.12] U/[ ml x g]) (P < 0.05), remarkably higher in the rats treated with Qiangjing ([130.23 ± 13.67] mg/[ml x g] [455 ± 51.50] U/[ml x g], and [18.56 ± 4.67] U/[ml x g]), Huangjingzanyu ([124.16 ± 14.02] mg/[ml x g], [ 419 ± 43.14] U/[ml x g], and [17.64 ± 4.08] U/[ml x g]), and levocarnitine ([123.34 ± 14.02] mg/[ml x g], [430 ± 31.80] U/ [ml x g], and [16.85 ± 5.55] U/[ml x g]) than in the models (P < 0.05). The Nrf2 mRNA expression was significantly reduced in the models as compared with the normal rats (P < 0.05) but remarkably increased in the Huangingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05). The SDH mRNA expression was significantly lower in the model than in the normal rats (P < 0.05) but markedly elevated in the Huangjingzanyu, Qiangjing and levocarnitine groups as compared with the model and normal animals (P < 0.05), remarkably higher in the Qiangjing than in the Huangjingzanyu group (P < 0.05).

CONCLUSIONOrnidazole induces OAS in rats, which is closely associated with excessive oxidation and energy metabolism dysfunction. Qiangjing Decoction can improve and even reverse ornidazole-induced OAS in rats as well as improve the ultrastructure of their testicular and epididymal tissues. Antioxidation and improvement of energy metabolism are probably the action mechanisms of Qiangjing Decoction in the treatment of OAS.

Animals ; Antioxidants ; Asthenozoospermia ; chemically induced ; drug therapy ; metabolism ; Carnitine ; pharmacology ; Disease Models, Animal ; Drugs, Chinese Herbal ; pharmacology ; Energy Metabolism ; drug effects ; Epididymis ; metabolism ; Glutathione Peroxidase ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Oligospermia ; chemically induced ; drug therapy ; metabolism ; Ornidazole ; Random Allocation ; Rats ; Sperm Count ; Sperm Motility ; Spermatozoa ; drug effects ; physiology ; Succinate Dehydrogenase ; metabolism ; Superoxide Dismutase ; metabolism ; alpha-Glucosidases ; metabolism