OBJECTIVETo construct mouse enhanced green fluorecence protein (EGFP) -peroxisome proliferator-activated receptor (PPAR)gamma2, and to detect EGFP-PPARgamma2 expression in infected mouse bone marrow mesenchymal stem cells (BMSC).

METHODSCut the fragment of PPARgamma2 from the expression plasmid pcDNA flag PPARgamma2, then cloned the gene fragment into pEGFP-C1 and pEGFP-N1 vector. Subsequently, subclone the fragment EGFP-PPARgamma2 from pEGFP-C1-PPARgamma2 into the shuttle plasmid DC315. HEK293 cells were co-transfected with the constructed recombinant shuttle plasmid DC315-EGFP-PPARgamma2 and large adenovirus helper plasmid pBHGlox deltaE1, 3Cre in mediation of liposome. The obtained replication-defective recombinant adenovirus Ad-EGFP-PPARgamma2 was confirmed. Then it was propagated in HEK293 cells. After the BMSC were transfected for 72 h, adipogenic differentiation was demonstrated.

RESULTSHEK293 cells were transfected with the pEGFP-C1-PPARgamma2 or pEGFP-N1-PPARgamma2 in mediation of liposome. The former green fluorescence protein was better than the latter by fluorescence microscope. The recombinant plasmids were digested and identified. Western blot analysis showed the expression of EGFP-PPARgamma2 in vitro. EGFP-PPARgamma2 protein was detectable in the nucleus of BMSC.

CONCLUSIONThe recombinant adenovirus encoding EGFP-PPARgamma2 fusion protein was successfully constructed, which provided a basis for application of EGFP-PPARgamma2 gene to adenovirus-mediated gene therapy.

Adenoviridae ; Animals ; Bone Marrow Cells ; metabolism ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; HEK293 Cells ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Mice ; PPAR gamma ; metabolism ; Recombinant Proteins ; metabolism ; Transfection

The chemical constituents of Taxus chinensis var. mairei cell cultures were investigated by chromatographic methods, including silica gel column chromatography, Sephadex LH-20 and preparative HPLC. Thirteen compounds were isolated from the 80% ethanol extract of cultured cells and their structures were elucidated by spectral data and physicochemical properties, which were identified as 2α,4α,7β,9α,10β-pentaacetoxy-14β-hydroxytax-11-ene (1), 2α,4α,7β,9α,10β-pentaacetoxytax-11-ene (2), 1β-deoxybaccatin VI (3), 2α-acetoxytaxusin (4), taxuyunnanine C (5), yunnanxane (6), 2α,5α,10β-triacetoxy-14β-propionyloxy-4 (20), 11-taxadiene (7), 2α,5α,10β-triacetoxy-14β-isobutyryloxy-4 (20), 11-taxadiene (8), 2α,5α,10β-triacetoxy-14β-(2'-methyl)butyryloxy-4 (20), 11-taxadiene (9), 13-dehydroxylbaccatin III (10), 13-dehydroxy-10-deacetylbaccatin III (11), paclitaxel (12) and (13) β-sitosterol. Among them, compound 1 is a new compound, and compounds 2, 4, 10 and 11 are isolated from the cell culture of Taxus chinensis var. mairei for the first time.
Alkenes ; analysis ; Cell Culture Techniques ; Cells, Cultured ; Diterpenes ; analysis ; Molecular Structure ; Paclitaxel ; analysis ; Sitosterols ; analysis ; Taxoids ; analysis ; Taxus ; chemistry

Animals ; Animals, Newborn ; Cyclic AMP Response Element-Binding Protein ; analysis ; Hippocampus ; blood supply ; chemistry ; pathology ; Hypoxia-Ischemia, Brain ; physiopathology ; Immunohistochemistry ; Proto-Oncogene Proteins c-fos ; analysis ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; physiopathology ; Time Factors

OBJECTIVETo observe the effect and safety of autologous cultured skin fibroblasts transplantation for treating depressed facial skin defects.

METHODSA total of 19 patients were treated from Jan, 2010 to Oct, 2010. Autologous skin fibroblasts were separated from postauricular skin biopsy or resected skin tissue in other surgeries such as blepharoplasty. They were cultured and expanded with exclusive method. Cells (2 x 10(7)/ml) within three passages were injected intradermally at the site of skin depression three times at one-month interval. Adverse events were observed and recorded. Clinical effects were evaluated and graded by two unrelated physicians before and 6 months after the first injection.

RESULTSCells from 16 patients were successfully cultured at the first time. The other 3 patients underwent a second harvest. A total amount of 6 x 10(8) cells could be reached within three passages in 45 days. 16 out of 19 patients accomplished the whole course of this study. Minor adverse events were observed in two patients including small ulcer caused by over injection in one patient and slightly redness and swelling in the other. The redness disappeared after a week without any treatment. No serious complications were observed. Significant difference was noticed between the scores obtained before and after the treatment.

CONCLUSIONSFrom this study, neither serious complications nor excessive cell proliferation or scar formation was found after cell injection. The effect of using autologous fibroblast transplantation was obvious and long-lasting, which provides a new choice for the treatment of depressed facial skin defects.

Adult ; Cells, Cultured ; Cicatrix ; therapy ; Face ; abnormalities ; Female ; Fibroblasts ; transplantation ; Humans ; Male ; Middle Aged ; Skin ; cytology ; Transplantation, Autologous ; Treatment Outcome ; Young Adult

Sixty healthy Sprague-Dawley male rats were used and divided randomly into control group (group C), cadmium loading group with medium dose (group M) and cadmium loading group with high dose (group H). Groups C, M and H were orally dosed daily with 0, 5 and 10 mg/kg of cadmium for over 6 weeks. Effects of cadmium loading on testis and endocrine function of reproduction in male rats were studied. The results showed that the zinc content decreased slightly in testis and plasma, and the cadmium concentration increased significantly in the testis of groups M and H; while the plasma levels of cadmium and zinc had no obvious difference as compared with those of group C; daily sperm production in the testis of group H decreased markedly during week 3 of cadmium loading, and was significantly lower in groups M and H as compared to that in group C during week 6; alkaline phosphatase (ALP) in group H and lactate dehydrogenase-X (LDH-X) in groups M and H were markedly lower compared to those of group C; plasma testosterone (T) level in both cadmium loading groups decreased and was low or significantly lower than that in group C; follicle stimulating hormone (FSH) and luteinizing hormone (LH) levels had no apparent difference between the three groups. It is suggested that the gradual accumulation of cadmium in testis tissue induced by chronic cadmium loading results in changes in some enzyme activity, a decrease in sperm production, and defect of endocrine function activity in the testis.
Alkaline Phosphatase ; drug effects ; Animals ; Cadmium ; blood ; Cadmium Chloride ; administration & dosage ; toxicity ; Follicle Stimulating Hormone ; blood ; Isoenzymes ; drug effects ; L-Lactate Dehydrogenase ; drug effects ; Luteinizing Hormone ; blood ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reproduction ; drug effects ; Spermatogenesis ; drug effects ; Testis ; enzymology ; pathology ; Testosterone ; blood

Computer Graphics ; Computer-Assisted Instruction ; Forensic Pathology ; Humans ; Imaging, Three-Dimensional ; Pathology ; education ; Pathology, Clinical ; Specimen Handling ; methods

OBJECTIVETo study the identification method of Pterocephalus hookeri.

METHODThe microscopical, Physicochemical and TLC methods were used.

RESULT AND CONCLUSIONThe convenient and effective identification methods for P. hookeri were established, which provide basis for its quality standard and development.

Chromatography, Thin Layer ; Drugs, Chinese Herbal ; analysis ; Magnoliopsida ; anatomy & histology ; chemistry ; Pharmacognosy ; Plant Leaves ; anatomy & histology ; chemistry ; Plant Roots ; anatomy & histology ; chemistry ; Plants, Medicinal ; anatomy & histology ; chemistry ; Quality Control

OBJECTIVETo explore the mRNA expression of KAI1 gene in laryngeal squamous-cell carcinoma and its clinical significance.

METHODSFresh laryngeal cancer samples taken from 40 laryngeal carcinoma cases and normal control laryngeal tissues from 9 subjects were examined with semi-quantitative reverse transcription polymerase chain reaction (RT-PCR).

RESULTSModerate, low and negative expression rates of KAI1 gene mRNA in nine normal laryngeal tissues were 33.3% (3/9), 33.3% (3/9) and 33.3% (3/9), respectively. The high, moderate, low and negative expression rates of KAI1 mRNA in 25 laryngeal cancers without lymph node metastasis were 40.0% (10/25), 28.0% (7/25), 20.0% (5/25) and 12.0% (3/25), respectively. The moderate, low and negative expression rates of KAI1 mRNA in 15 laryngeal cancers with lymph node metastasis were 20.0% (3/15), 26.7% (4/15) and 53.3% (8/15), respectively. The KAI1 mRNA expression in the laryngeal cancers without lymph node metastasis was higher than that in normal laryngeal tissues (P < 0.05). The KAI1 mRNA expression in the laryngeal cancers with lymph node metastasis was lower than that in the laryngeal cancers without lymph node metastasis (P < 0.05). The high, moderate and low expression rates of KAI1 mRNA in 10 highly differentiated laryngeal cancers were 50.0% (5/10), 30.0% (3/10) and 20.0% (2/10), respectively. The high, moderate, low and negative expression rates of KAI1 mRNA in 12 low differentiation laryngeal cancers were 8.3% (1/12), 16.7% (2/12), 16.7% (2/12) and 58.3% (7/12), respectively. The differences of KAI1 mRNA expression between high and low differentiation laryngeal cancers were statistically significant (P < 0.05).

CONCLUSIONThe decrease of KAI1 mRNA expression may be related to lymph node metastasis and low differentiation of laryngeal squamous-cell carcinoma.

Adult ; Aged ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Female ; Humans ; Kangai-1 Protein ; biosynthesis ; genetics ; Laryngeal Neoplasms ; genetics ; metabolism ; pathology ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Objective To systemically evaluate the effect of protective analgesia on preventing phantom limb pain(PLP)after amputa-tion.Methods Published articles from the earliest date available to June,2017 were recalled from Cochrane Library,PubMed,Embase,Web of Science,OVID,and Science Direct to collect prospective studies using protective analgesia in perioperative period to prevent PLP after amputation.Two reviewers screened literatures referring to studies according to the inclusion and exclusion criteria,and assessed the quality of them.Data of general information and incidence of PLP in the follow-up period were extracted and analyzed with RevMan 5.3 software. Results Six studies were included with a total of 256 patients in the one-month follow-up period including 127 cases in the protective anal-gesia group(group P)and 129 cases in the control group(group C),a total of 232 patients in the six-month follow-up period including 114 cases in group P and 118 cases in group C,and a total of 118 patients in the twelve-month follow-up period including 58 cases in group P and 60 cases in group C.The incidence of PLP were lower in group P than those in group C in the one-month follow-up period(RD=-0.21, 95%CI[-0.38,-0.04],Z=2.47,P=0.01)and in the six-month follow-up period(RD=-0.28,95%CI[-0.52,-0.05],Z=2.37,P=0.02),and it was not significant in the twelve-month follow-up period(RD=-0.20,95%CI[-0.48,0.09],Z=1.35,P=0.18).Conclusion Protective analge-sia in perioperative period can prevent against PLP after amputation in the recent time,however,it needs further observation in long-term.

OBJECTIVETo investigate the effect of p38 mitogen-activated protein kinase (p38MAPK) on the expression of COX-2 and caspase-3 in the substania nigra (SN) of mice with MPTP-induced Parkinson disease (PD).

METHODSC57BL/CN mice were treated with MPTP to prepare a subacute PD model, and their behavioral changes following the treatment were observed. Immunohistochemistry and Western blotting were performed to detect the expression of tyrosine hydroxylase (TH), COX-2 and phosphorylation of P38MAPK in the SN and their changes following treatment with SB203580, a specific inhibitor of P38MAPK.

RESULTSThe 7-day model group showed typical symptoms of PD with decrements of TH-positive neurons and TH protein level in the SN of the midbrain by about 65% and 75%, respectively (P<0.01). In the 3-day model group, the COX-2-, caspase-3- and phosphorylated P38MAPK-immunoreactive cells and their protein levels in the SN increased markedly with obvious loss of TH-positive neurons. Administration of SB203580 obviously lessened the above changes (P<0.01).

CONCLUSIONP38MAPK regulates the inflammation and apoptosis in the SN of the mouse model of subacute PD, and SB203580 may provide some neuroprotective effect.

Animals ; Caspase 3 ; genetics ; metabolism ; Cyclooxygenase 2 ; genetics ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Parkinson Disease ; metabolism ; Signal Transduction ; Substantia Nigra ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism