[ABSTRACT]AIM:ToinvestigatetheroleoftinyantisensenucleicacidagainstmiR-155(tinyantimiR-155, t-antimiR-155) in multiple myeloma cells .METHODS:According to the seed sequence of miR-155, t-antimiR-155 was designed and synthesized .t-antimiR-155 was transfected by Lipofectamine TM 2000 into RPMI-8266 cells.The cells were di-vided into t-antimiR-155 group, scrambled control (SCR) group and blank control group .The growth-inhibitory potencies were measured by MTT assay .The ability of cell colony formation was detected by cell colony formation assay .The cell ap-optosis was assessed by flow cytometry with annexin V /PI double staining .RESULTS: The best concentration and time were 0.4 μmol/L and 48 h, respectively.The cell colony forming experiment showed that the circumstances of forming cell community in t-antimiR-155 group was weaker than that in SCR group , and the colony formation inhibitory rate of former was significant higher than the latter .Compared with SCR group , the cell apoptosis in t-antimiR-155 group significantly in-creased.CONCLUSION: The t-antimiR-155 inhibits the progression of multiple myeloma cells by interfering with miR-155.miR-155 may serve as a potential target in gene therapy for treating multiple myeloma .

Objective To construct the eukaryotic expression plasmids of short hairpin RNA (shRNA) specific for mouse Bcl-2-assoeiated athanogene 1 (BAG-1) and to observe their inhibitory effects on the expression of BAG-1 gene in mouse melanoma B16FI0 ceils. Methods Plasmids named pRNAT-U6.1/Neo-BAG-1, were designed and constructed to target the mouse BAG-1 mRNA coding region. LipofectaminTM 2000 was used to transfect plasmids into BI6F10 cells. Negative plasmid-transfected and tmtransfected B16F10 cells served as negative and blank controls respectively. Forty-eight hours following transfection, G418 was used to select the resistant cells. The mRNA and protein expression of BAG-1 gene was measured by reverse transcription-PCR and Western blot respectively about 1 month after the transfection. Results The eukaryotic expression plasmids, pRNAT-U6.1/Neo-BAG-1, were constructed, and verified by restriction enzyme digestion and DNA sequencing. The transfection rate in B16F10 cells was 20% -30%. Compared with the blank control, the mRNA and protein expression of BAG-1 in BI6FI0 cells was significantly inhibited by BAG-1 shRNA (both P<0.05), and the inhibition rates were (77±4)% and (62 ±2)%, respectively. Conclusions These results indicate that the eukaryotic expression vectors containing shRNA against BAG-1 gene, pRNAT-U6.1/Neo-BAG-1, are successfully constructed, and can significantly inhibit the expression of BAG-1 gene in mouse melanoma B16F10 cells.

Objective To explore the operative methods and clinical outcomes in emergency or sube- mergency repair for the complicated tissue defects in hand in first stage applying microsurgical technique. Methods From Jan.,2000 to Aug.,2005,49 emergency cases of complicated hand tissue defects were re- paired in the first stage with replantation,reconstruction,free flaps,combined finger reconstruction and flap transplantation,including 21 cases in mini tissue mass replantation or reconstruction,15 cases in replantation combined with free flap transplantation,8 cases in replantation and reconstruction combined with free flap transplantation,5 cases in combined multiple digits reconstruction with free flap transplantation.The free flap transplantation included the anterolateral femoral flap,the latissimus dorsi myocutaneous flap,the dorsalis pe- dis flap,the media pedis flap and the instep flap.Results All the flaps,the replanted and reconstruceted finger survived uneventfully except for one replanted finger necrosis.45 cases healed in the first stage and the other 4 cases healed in the second stage.During a follow-up from 6 months to 3 years postoperatively,a satis- factory appearance and function of the reconstructed hand was achieved.The excellent and good rate was 85.7% assessed with provisional functional assessment criterion for upper limbs issued by Chinese Society of Hand Surgery.Conclusion The emergency or subemergency repair for the complicated tissue defects in hand has the advantage of short-term treatment and desirable functional outcome.The emergency replantation and reconstruction combined with various flaps or tissue mass can be applied to repair tissue defect in hand in the first stage according to the position and area of the defect along with the technique level of the surgeon, having been proved to achieve desirable clinical outcomes.And the key points leading to a successful operation is the correct treatment for the raw surface of the defects,suitable choice for various flaps,logical design of combination pattern and prevention and timely treatment for vessel crisis.

Objective:To evaluate the effect and mechanism of davallia mariesil flavones (DMF) improving osteoporosis via Notch signaling pathway.Methods:(1) 120 cases patients with osteoporosis in our hospital from January 2016 to January 2017 were analyzed,and divided into experiment group (60 cases) and control group (60 cases) using digital random grouping method.Experiment group was treated with DMF combined with D-calfor.Control group was treated with D-calfor only.After treatment, the levels of serum calcium,serum phosphor,TNF-α,MCP-1 and IL-6 in serum were detected to evaluate the clinical efficacy.(2) Bone marrow stromal cells were separated and cultivate.NC group:DMEM(contain 10% FBS).RA group:RA(0.4 mmol/L).DMF+RA group:DMEM(contain DMF)+RA(0.4 mmol/L).Jaggedl+RA group:Jaggedl(1 000 μg/L)+RA(0.4 mmol/L).Jaggedl+DMF+RA group:Jaggedl(1 000 μg/L)+DMEM(contain DMF)+RA(0.4 mmol/L).DAPT+RA group:DAPT(16 μmol/L)+RA(0.4 mmol/L). DAPT+DMF+RA group:DAPT(16 μmol/L)+DMEM(contain DMF)+RA(0.4 mmol/L).Western blotting assays and PCR were performed to assess mRNA and protein levels of Notch-1,Hes-1.Results: (1) In clinical study,the effective rate in treatment group was obviously higher than control group (91.67%>76.67%,P<0.05).The levels of serum calcium and serum phosphor in the experiment group was higher than in the control group (P<0.05).The levels of TNF-α,MCP-1 and IL-6 in the experiment group was lower than in the control group (P<0.05).(2) In experimental study,compared with the RA group,the expressions of Notch-1,Hes-1 mRNA and protein were upregulated in Jaggedl+RA group,but were downregulated in DAPT+RA group,DMF+RA group (P<0.05). Compared with the Jaggedl+RA group,the expressions of Notch-1,Hes-1 mRNA and protein were downregulated in Jaggedl+DMF+RA group (P<0.05).Compared with the DAPT+RA group,the expressions of Notch-1,Hes-1 mRNA and protein were downregulated in DAPT+DMF+RA group (P<0.05).Conclusion:DMF could improve the condition of osteoporosis.The mechanism may be associated with inhibiting the Notch signaling pathway by DMF.

[Objectives]To investigate the protective effects of recombinant Trichinella spiralis excretory-secretory 53ku pro-tein(rTsP53)on acute lung injuries in mice.[Methods]Thirty Balb/c mice were randomly divided into normal group. ALI group and rTsP53 group(n=10,respectively). Macrophages were harvested by bronchoalveolar lavage. Mortality in 72 hours was counted and compared. Pathological damage of lung tissues was observed by HE staining and graded by Smith score. Wet/dry ratio was measured. The bronchoalveolar lavage fluid concertration of IL-6 and IL-4 was measured by ELISA. The mRNA expression of TNF-α,iNOS, IL-10 and Arg-1 in alveolar lavage macrophages was detected by RT-PCR.[Results]72 h mortality of ALI mice was 70%,which was reduced to 30% in mice received rTsP53 treatment. Compared with ALI mice,the pathological damage of in rTsP53 treated-mice was improved and Smith score was declined ,combined with descending W/D ratio. IL-6 level of alveolar lavage fluid was elevated in ALI mice compared with normal group. And alveolar lavage macrophage was polarized to M2 sub-type,appeared as higher mRNA expression of TNF-α and iNOS and lower level of IL-10 and Arg-1. Bronchoalveolar lavage fluid concentration of IL-6 was declined and IL-4 was elevated in rTsP53-treated mice compared with ALI group. The macrophages of alveolar wash had higher mRNA expression of IL-10 and Arg-1,while lower level of TNF-α and iNOS,manifesting M2 polarization characteristics.[Conclusion]Recombinant T.spiralis P53 protein could protect mice from acute lung injuries induced by LPS via modulating M2 macrophage polarization,which play a role in depression of inflammatory reaction and tissue repairment.

Novel dosage forms emerges more and more in recent years. One of them is liquid-filled hard gelatin capsules, which adopt gelatin or the hydroxypropyl methyl cellulose (HPMC) as capsule shell. The liquid-filled hard gelatin capsule is increasingly getting attention because of its new-concept dosage form design, which deliver liquid drugs by solid form. The paper mainly introduces application, pharmaceutical manufacturing, quality assessment, prospect of liquid-filled hard gelatin capsules, and focuses on the application and pharmaceutical manufacturing (preparation) of liquid-filled capsule. It is suggested that the capsule is suitable for various liquid or semi-solid natural plant extract and achieve different release profiles. The preparation adopted liquid-filled hard capsules technology. The influence factors concluded property of shell and device of filling. The quality was often evaluated by moisture content of capsule shell, dissolution rate etc. At the same time, it was pointed out that the new dosage form has remarkable marketing prospect and bring profits for enterprises.
Capsules ; chemistry ; Drug Delivery Systems ; Drugs, Chinese Herbal ; Gelatin ; chemistry

OBJECTIVETo employ reverse first dorsal metatarsal artery island flap to recover the donor site of great toe after free lateral pulp flap transfer, and decrease the morbidity of microsurgical donor site.

METHODSFrom February 2000 to June 2004, 12 cases of soft tissue defect in thumb and finger were treated by free lateral pulp flap of great toe transplantation, and that the secondary defect of great toes were repaired by reverse first dorsal metatarsal artery flap according to the anatomical communicating between first dorsal metatarsal artery and plantar metatarsal artery. The donor exposure of dorsal pedis were straightly sutured.

RESULTSAll the lateral pulp flaps of great toe and the reversed first dorsal metatarsal flaps survived uneventfully with desirable appearance and sensation. In thumb and finger defect reconstructed with lateral pulp flaps of the great toe, there was in mean static two-point discrimination of 6 mm at 10 month follow-up, that was 10 mm in reverse flaps.

CONCLUSIONSIt is an instructive and practical creation to adopt the reverse first dorsal metatarsal artery flap to recover the raw surface in the lateral aspect of the great toe, which minimizes the morbidity at the donor site and gains the twin-win results.

Adolescent ; Adult ; Arteries ; transplantation ; Female ; Finger Injuries ; surgery ; Humans ; Male ; Metatarsus ; blood supply ; Middle Aged ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; pathology ; Young Adult

Objective:To discuss the topical action characteristics of the biological transmission of moxibustion heat via temperature collection and numerical modeling.Methods:Temperature of moxibustion was measured at multiple points at a distance of 3 cm to obtain the moxibustion temperature field nephograms by the high-accuracy temperature measure array.Finite element analysis was used to imitate the three-dimensional dynamic distribution of temperature in acupoint tissues.Results:Through numerical analysis,the one-dimensional,two-dimensional and three-dimensional distributions of temperature in human acupoint tissues at 5 min of moxibustion were established.The result showed that moxibustion heat mainly transmitted from the surface of the tissue to the internal,and the influence of moxibustion heat decreased with the depth of the tissue.The analysis of the nephograms of acupoint tissue temperature at 5,10,15 and 20 min of moxibustion showed that with the increase of the moxibustion time,the temperature in acupoint tissues constantly rose,and the transmission depth of moxibustion heat also further expanded inside acupoint.Conclusion:By establishing the three-dimensional dynamic model of heat transmission inside acupoint tissues with the biological parameters of human tissues and the temperature values obtained,this study used finite element analysis software ANSYS 14.0 and discovered the rules in the transmission of heat in body tissues during moxibustion,and the features in moxibustion heat transmission (from the proximal to the distant) and heat penetration (from the surface to the internal).This study provides theoretical and experimental support for the application of moxibustion in clinical practice.

Objective To investigate the relationship between the clinical manifestations,prognosis and histopathological findings of mucormycosis.Methods The clinical data on and pathological findings from 7 cases of mucormycosis confirmed by fungal culture in the institute from 1989 to 2006 were analyzed retrospectively.Results There was 1 case of hinocerebral mucormycosis and 6 cases of cutaneous mucormycosis,among them,2 were mucormycotic necrotizing fasciitis (MNF).The condition of patients with rhinocerebral mucormycosis or MNF aggravated rapidly and all the 3 patients died from mucormycosis. Histopathological examination showed mixed infiltrates of inflammatory cells as well as necrosis and angioin vasion.On the contrary,the condition of the remaining 4 patients with cutanesus mucormycosis,who presented mainly with indurated erythematous patch,progressed slowly,and 2 patients were cured.Histologically,the lesions were characterized by granulbmatous infiltration with a few hyphae;no typical angioinva sion phenomenon was noted.There was no evidence of perineural invasion with hyphae in any of the 7 cases.ConclusionIn patients with mucormycosis,histopathological findings characterized by mixed infiltrates of inflammatory cells,numerous hyphae and typical angioinvasion phenomenon may herald a poor prognosis.

OBJECTIVETo establish 2-dimensional gel electrophoresis (2-DE) images and seek differentially expressed serum proteins for understanding the pathogenesis of silicosis.

METHODS2-DE and matrix-assisted laser desorption/ionization time of flight tandem mass spectrometry (MALDI-TOF-MS/MS) were used to screen differentially expressed serum proteins among silica-exposed population, suspect of silicosis (0+), phase one (I) group with silicosis and control group(non silica exposure).

RESULTSComplement C4, leucine-rich alpha-2-glycoprotein and alpha-1-antitrypsin were significantly highly expressed in suspect of silicosis (0+) group(P < 0.01), but lowly in other groups. Inversely, serotransferrin was significantly down-regulated only in suspect of silicosis (0+) group(P < 0.01). Plasma glutathione peroxidase, tetranectin, apolipoprotein A-I and transthyretin were equally expressed in the serum of control group and silica-exposed population group, but decreased in the suspect of silicosis (0+) and phase (I) group.

CONCLUSIONComplement C4, leucine-rich alpha-2-glycoprotein, alpha-1-antitrypsin, serotransferrin, plasma glutathione peroxidase, tetranectin, apolipoprotein A-I and transthyretin are differentially expressed in the silica-exposed group and phase (I) group with silicosis, and the result should be validated by other biochemical technologies.

Adult ; Aged ; Aged, 80 and over ; Blood Proteins ; analysis ; Electrophoresis, Gel, Two-Dimensional ; Humans ; Middle Aged ; Proteomics ; methods ; Silicosis ; blood ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization