Objective To prepare recombinant adenovirus pAd-gal-9 containing murine galectin-9 and explore galectin-9's pro-apoptotic effect on T lymphocytes. Methods The recombinant adenovirus plas-mid pAd/CMV/V5-DEST-gal-9 was prepared by conventional molecular cloning and LR reaction. The pAd/ CMV/V5-DEST-gal-9 linearlized by Pac I was transfected into 293A cells with Lipofectin 2000. Eight days after transfection, the 293A cells were subjected to freeze/thraw circle for three times and the supernatant was collected after centrifugation. Higer titer pAd-gal-9 was produced by large-scale infection of 293A cells with the supernatant containing pAd-gal-9. The supernatant was condensed to get purified pAd-gal-9 by CsCl density gradient centrifugation. After titer determination with gradient dilution of harvested pAd-gal-9 infec-tion in 293A-seeded 96-wells, pAd-gal-9 was used to infect the CHO cell line. Immunohistological assay, Western blot and flow cytometry were employed to ascertain the subcellular location expression of galectin-9. We added solid-phase transgenic CHO cells or freshly-cultured supernatant to medium containing activated T cells to detect the pro-apoptotic effect of galectin-9. Results The pAd-gal-9 was prepared successful. Im-munohistochemical staining of CHO infected with pAd-gal-9 confirmed that galectin-9 was expressed in the cytosol. Intercellular staining indicated that mean fluorescence intensity of galectin-9 was significantly higher in pAd-gal-9-infected CHO group than control group. Supernatant from pAd-gal-9-infected CHO promoted the apoptosis of T cells. The percent of apoptotic T cells was higher than the Tim-3 positive T cells. Conclu-sion CHO infected with pAd-gal-9 can secret galectin-9 to promote the apoptosis of activated T cells via Tim-3-independent mechanisms.

Objective To summarize the clinical features of Chinese patients with mitochondrial encephalomyopathy, lactic acidosis and stroke-like episodes ( MELAS).Methods A total of 190 patients with MELAS who presented to Peking University First Hospital between 1997 and 2015 were recruited.Among 190 patients, 175 were identified carrying mitochondrial DNA mutations, and the remaining 15 patients were diagnosed by muscle biopsy.The clinical features, including predisposing factors of stroke-like episodes, the onset symptoms and frequencies of various manifestations were analyzed and reported.Results In our cohort of MELAS patients, the male-to-female ratio was 1.44∶1.The median age of onset was 14 years ( from 7 months to 45 years).The peak onset ages were 8-12 years.The median onset age of the first stroke-like episode was 16 years ( from 1 to 53 years ).There were 66 ( 46.15%) patients who had predisposing factors before the onset, and fatigue and upper respiratory tract infection were the most common predisposing factors of stroke-like episodes in these patients ( 37.88%, 25/66 and 34.85%, 23/66, respectively).Other predisposing factors included emotional agitation, drinking alcohol, trauma, withdrawal of antiepileptic drugs, being frightened, satiation and hunger.Stroke-like episodes appeared in 70.53%(134/190) patients as an onset symptom and developed in all patients with disease progression.The neurological manifestations included seizure ( 89.42%, 169/189 ) , mental retardation or dementia (82.87%, 150/181), headache (74.30%, 133/179), hemianopia or cortical blindness (67.72%, 107/158), exercise intolerance (50.87%, 88/173), hemiplegia or hemianesthesia (47.44%, 74/156), sensorineural deafness (46.20%, 85/184), aphasia (39.47%, 60/152), behaviour disorder (17.71%, 31/175) and ophthalmoplegia ( 9.60%, 17/177 ).The manifestations of extra-nervous systems included hirsutism (67.57%, 100/148), vomiting (65.58%, 101/154), fever (62.07%, 90/145), short stature (45.32%, 63/139), diarrhea or constipation (43.48%, 70/161), low body mass index (26.62%, 37/139), diabetes mellitus (20.79%, 37/178) and kidney disease (3.16%, 6/190).Conclusions The majority of the patients in this study have the disease onset during childhood.There are more male MELAS patients than females.Most common clinical manifestations are seizure, mental retardation or dementia, headache, cortical blindness, hirsutism, vomiting and fever in this patient group.

OBJECTIVETo investigate the hard and soft tissue changes following the treatment of Class II division 1 malocclusion using Twin-block appliance.

METHODS50 Class II division 1 malocclusion subjects whose handwrist radiographs were in FG-G stage were selected. One group (27 patients) was treated with Twin-block appliance, the other group (23 patients) was observed without treatment The acquired data of cephalometric of two groups were analyzed statistically with SPSS 11.0.

RESULTSSoft tissue changes, Ls-E, Li-E, U1-Stms, Stms-Stmi, NsLs-FH, LsNsLi, LsNsPg', the angle of H decreased. Sn-Stms, Stmi-Me', Ns-Me', Sn-Me', NsLi-FH, NsPg'-FH, A'Ls-FH, B'Li-FH, LiB' Pg', CmSnLs, GSnPg', the angle of Z increased, there was statistically significant difference (P < 0.05). Hard tissue changes, SNB, L1-NB, IMPA increased, ANB, U1-SN, U1-NA, FMIA decreased, there was statistically significant difference (P < 0.05).

CONCLUSIONSagittal relationship between upper- and lower-jaws is effectively improved after orthopedics with Twin-block appliance. Lower face height increases. Soft tissue profile tends to be straight-styled.

Cephalometry ; Face ; Humans ; Malocclusion ; Mandible

Objective To explore the effects of traditional Chinese medicine ( TCM) nursing on the acute stage of stroke ( cerebral infarction) in clinical application. Methods A total of 67 patients with acute cerebral infarction were in strict accordance with the implementation of nursing programs and TCM. The nurses who were responsible for the implementation of the nursing program of TCM were received hospital uniform training and assessment. We collected the main symptoms of patients during hospitalization, the implementation of the main syndrome nursing methods and TCM nursing technology, as well as the main symptoms of improvement. Results For patients′ main symptoms ( consciousness disorder and hemiplegia, dizziness and headache, high fever, sputum interest promotion, two fecal incontinence, constipation, sluggish speech and swallowing) , the dialectical nursing implementation rate was 100%; 10 kinds of TCM nursing technology applied, can effectively improved the patients′ hemiplegia, dizziness, headache, constipation, swallowing difficulties and other symptoms. Conclusions Stroke ( cerebral infarction) of TCM nursing program in acute stage can promote the development of clinical TCM nursing, play an effective TCM nursing technology advantages, and is worthy of further promotion and application.

Objective: To explore the importance and necessity that central sterile supply department (CSSD) carried out detection for insulating property of electrosurgical instrument so as to guarantee the safety of surgery for patients. Methods: 50 monopole instruments and 30 bipolar instruments were collected, and specific insulation tester was adopted to detect the insulation property of these instruments. Besides, the detection methods of electrosurgical instrument between home and abroad were compared so as to avoid and reduce the adverse events caused by the breakage of insulating layer of electrosurgical instrument. Results: The breakages of insulating layer of 5 monopole instruments and 2 bipolar instruments were found through detection and all of them were replaced. The same problem did not be found again after 2 months. The insulation tester was used to detect the insulation property of electrosurgical instruments could effectively found these instruments with damages of insulating layer. Conclusion:The damage of the insulation layer of the electrosurgical instruments may cause serious consequences and even threat safety of patients. Periodic detection for insulation property can effectively reduce and avoid the occurrence of same problems and guarantee the safety of patient. Therefore, it is very necessary measurement.

OBJECTIVETo research (CAG)n polymorphism of androgen receptor gene in idiopathic azoospermic and oligospermic Chinese men, and to explore its correlation with the development of dyszoospermia.

METHODSFifty-two men with oligospermia and 31 with azoospermia were enrolled, the number of CAG sequence repeats determined by PCR and denaturing polyacrylamide gel electrophoresis (PAGE), and (CAG)n polymorphism analyzed.

RESULTSThe CAG repeats in the men with oligospermia and those with azoospermia were 22.19 and 22.13 respectively.

CONCLUSIONThe polymorphism of the repeat length of the (CAG) n satellite in androgen receptor gene may affect spermatogenesis and have a less important role in male infertility.

Adult ; Electrophoresis, Polyacrylamide Gel ; Exons ; Humans ; Infertility, Male ; genetics ; Male ; Oligospermia ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Receptors, Androgen ; genetics ; Trinucleotide Repeats ; genetics

OBJECTIVETo investigate the effects of GM1 on inducing adult rat bone marrow stromal cells (MSCs) to form neural progenitor cells and their differentiation.

METHODSPurified MSCs were induced by different components of basic fibroblast growth factor (bFGF) alone, GM1 alone or combination of bFGF with GM1. After 3 days' incubation, fibronectin and collagen I were detected with immunocytochemistry, and nestin was detected with immunofluorescence. Neuron-specific enolase (NSE), glial fibrillary acidic protein (GFAP) and galactose cerebroside (GalC) were detected with immunocytochemistry after 7 days' incubation.

RESULTSAfter induction with bFGF alone or combination of bFGF and GM1, some MSCs exhibited the phenotypes of neural progenitor cells, and then neurons and astrocytes. In these two groups, the positive cells for fibronectin and collagen I decreased markedly after 3 days' induction. At the same time, the positive cells for nestin increased markedly. After 7 days' induction, NSE and GFAP-positive cells increased significantly. Furthermore, the addition of bFGF and GM1 caused the maximal variation. However, addition of GM1 alone had no inductive effects.

CONCLUSIONSCombination of bFGF with GM1 may synergistically promote the transformation of MSCs and differentiation into neurons and astrocyte-like cells. The results suggest a promising route for the application of MSCs.

Analysis of Variance ; Animals ; Bone Marrow Cells ; Cell Differentiation ; drug effects ; physiology ; Cells, Cultured ; Drug Synergism ; Fibroblast Growth Factor 2 ; pharmacology ; Fluorescent Antibody Technique ; G(M1) Ganglioside ; pharmacology ; Immunohistochemistry ; Probability ; Rats ; Rats, Wistar ; Sensitivity and Specificity ; Stem Cells ; pathology ; physiology ; Stromal Cells ; drug effects ; physiology ; ultrastructure

Objective:To investigate the inhibitory effect of in vivo non-targeting transfection of recombinant fibronectin polypeptide CH50 against tumors and to study the related mechanisms.Methods:After inoculated with tumor cells, BALB/c mice were injected with CH50 plasmids,control plasmids,and normal saline separately.The growth of the tumor was observed;the expression of genes (such as B7-1,B7-H1 etc.) in tumor tissues was detected by RT-PCR;and the count of T lymphocytes in local tumor tissues was analyzed by flow cytometry.Results:The tumor growth was obviously suppressed by in vivo CH50 expression.The expression of genes (B7-1 and B7-H1) was up-regulated along with the growth of tumor.CH50 increased the ratios of B7-1/B7-H1 and B7-1/B7-DC and suppressed the up-regulation of IL-10 and TGF-?genes.The direct action of CH50 on H22 cells resulted in the down-regulatoin of TGF-?gene.The count of T lymphoeytes in tumor tissues of CH50 treatment group was significantly higher than that in other groups.Conclusion:Ex- pression of CH50 by non-targeting transfection can effectively inhibit the growth of tumor;the regulation of the immuno- regulatory genes in tumor mieroenvironment is an important part of the treatment mechanism of CH50.

Bombyx mori nucleopolyhedrovirus (BmNPV) bm47 gene is found in all sequenced lepidopteran nucleopolyhedroviruses (NPVs). It is one of the core genes of NPVs. However, the role of bm47 in the biological cycle of NPV remains unknown. In this study, the Red recombination system was used to knock out bm47 from BmNPV to construct bm47-ko-Bacmid in E. coli BW25113 system. Then bm47 gene was introduced back to the viral genome using the Bac-to-Bac system to create the repair virus bm47-re-Bacmid. TCID50 assay and real-time PCR (qPCR) were used to evaluate the effects of bm47 deletion on viral DNA replication, gene transcription, and protein expression. qPCR results showed that bm47 knock-out had no significant effect on viral DNA replication. However, the qPCR results showed that bm47-ko-Bacmid significantly decreased the transcription levels of early gene lef-3, late gene vp39, and very late gene p10 at 48 h and 72 h after viral transfection of BmN cells (P < 0.05). This work will provide a foundation for further studies on the biological function of BmNPV bm47 in viral replication and transcription.
Animals ; Bombyx ; virology ; Gene Deletion ; Gene Expression Regulation, Viral ; Nucleopolyhedrovirus ; genetics ; physiology ; Transcription, Genetic ; Viral Proteins ; genetics ; metabolism ; Virus Replication

OBJECTIVETo investigate the role and significance of cardiac mast cells and Toll-like receptor 4 (TLR4) in the development and progression of viral myocarditis (VMC).

METHODSForty-eight Balb/c mice were randomly divided into a control group (n=24) and a model group (n=24). Coxsackievirus B3 was intraperitoneally injected into the model group mice to establish a VMC model. In each group, cardiac tissues were collected from 8 mice at 7, 14 and 28 days after the model was established. The cardiac tissues were stained with hematoxylin and eosin as well as Masson trichrome to observe pathological changes in cardiac tissues. The number and degranulation of cardiac mast cells at each time point were measured and evaluated by toluidine blue staining and transmission electron microscopy. The mRNA and protein expression of TLR4 in cardiac tissues was measured by RT-PCR and immunohistochemistry. In the model group, the correlation between number of cardiac mast cells and mRNA expression of TLR4 at all time points was analyzed.

RESULTSThe model group had significantly higher pathological scores of cardiac tissues than the control group at all time points (P<0.05). The myocardial collagen volume fraction in the model group at 28 days was significantly higher than in the control group at all time points and higher than in the model group at 7 and 14 days (P<0.05). At each time point, the model group had a significantly increased number of mast cells (P<0.05), and significantly increased mRNA and protein expression of TLR4 (P<0.05) compared with the control group. In the model group, the number of cardiac mast cells was positively correlated with the mRNA expression of TLR4 at all time points (R²=0.877, P<0.05).

CONCLUSIONSMice with VMC have significantly increased numbers of cardiac mast cells and expression of TLR4 compared with control mice at all time points, suggesting that mast cells and TLR4 may play important roles in the inflammatory response and fibrosis of VMC.

Animals ; Coxsackievirus Infections ; immunology ; Enterovirus B, Human ; Female ; Mast Cells ; physiology ; Mice ; Mice, Inbred BALB C ; Myocarditis ; immunology ; Myocytes, Cardiac ; pathology ; Toll-Like Receptor 4 ; analysis ; genetics ; physiology