Animals ; Antibodies, Anti-Idiotypic ; therapeutic use ; Disease Models, Animal ; Doxorubicin ; toxicity ; Immunotherapy ; Interleukin-18 ; immunology ; pharmacology ; Male ; Mice ; Nephrosis, Lipoid ; chemically induced ; pathology ; therapy

Objective To prepare a new type of micropore porcine acellular dermis matrix with the aid of laser (LPADM),and to validate the healing effect of the LPADM through the phrase Ⅰ composite transplanting on the back of the full- thickness skin defects in SD rats.Methods In vitro,the allogeneic fibroblasts were separately cultured with the LPADM (LPADM group) or the non-pore PADM (non-pore LPADM group),while fibroblasts cultured by pure medium were used as control.After culture of 1 day,3 days and 5 days,the contents of IL-10,IL-6,TGF-β1,LN,VEGF expressed by fibroblasts were determined by double-antibody sandwich ELISA method.In vivo,the phrase Ⅰ transplantations of LPADM graft with split-thickness autologous skin were carried on the backs of the full-thickness cutaneous defects of SD rats.The healing condition was observed and analyzed by histological tests.Results The differences of the absorbance value between the LPADMgroup,PADM group and control group in each day were not statistically significant (F=0.050-1.763,P＞0.05).The transplantation of LPADM graft with split-thickness autologous skin graft resulted in high rate of surviving without signs of rejection 3 weeks later.After 1-month of transplantation,the regenerated skin was well enough to be lifted without any serious scars.Conclusions The phrase Ⅰ transplantation of LPADM graft with split-thickness autologous skin graft can accelerate the healing process of full-thickness skin wounds with high biological safety.

Objective To study the value of high frequency ultrasound in predicting spontaneous closure of patent duct arteriosus (PDA) in premature neonates. Methods One hundred and seventy premature neonates aged less than 37 weeks of gestation were enrolled. They were all admitted in the neonatal intensive care unit of Jinhua People′ s Hospital within 24 hours after birth. Echocardiographic examinations was conducted using GE′s Vivid I Portable color Doppler ultrasonography with routine cardiac probe (frequency 1.7-3.4 MHz) and high-frequency transducer (frequency 7.5 MHz). Up to the 7th day, the echocardiography was carried out every day after birth until the closure of PDA .They were divided to two groups according to whether spontaneous closure of PDA took place with in 7 days after birth (the control group) or not (the PDA group). There were 16 premature neonates in PDA group, including 6 male and 10 female. The mean gestational week at birth was (35.3±1.4) weeks. The mean birth weight was (2 330.9±325.5) g. Thirty-two premature neonates were included incontrol group, including 13 male and 19 female. The mean gestational week at birth was (35.0±1.3) weeks. The mean birth weight was (2 337.5±287.8) g. The clinical characteristics and echocardiographic parameters were compared between the two groups. Results The detection rates of high-frequency ultrasound and routine echocardiography were all 100%. The spontaneous closure rate of PDA in preterm infants at 24 hours, 48 hours, 72 hours were 18.8%(32/170), 61.2%(104/170), 78.8%(134/170) respectively. Within 7 days after birth, the closure rate was 90.6%(154/170), therefore the incidence of PDA in premature neonates was about 9.4%(16/170) in our study. The display rate of entire duct arteriosus using high frequency ultrasound was signiifcantly higher than that using routine ultrasound [82%(39/48) vs 46%(22/48), P<0.001, 100%(48/48) vs 77.1%(37/48), P<0.01]. The diameters of duct arteriosus in PDA group were signiifcantly larger than that in control group [(2.08±0.4) mm vs (1.09±0.22) mm, P<0.001]. However, the remaining echocardiographic parameters showed no signiifcant difference between the two groups (P>0.05). To predict the spontaneous closure of PDA in preterm neonates within 7 days after birth by high frequency ultrasound, the sensitivity, specificity and accuracy were 87.5%, 90.6% and 66.5% respectively using the cutoff point of 1.55 mm. While the sensitivity, speciifcity and accuracy were 75%, 93.7%, 44.15%by routine echocardiography with the cutoff point of 2.36 mm. Conclusions High-frequency ultrasound can conifrm the diagnosis of PDA in preterm infants and it is better than routine echocardiography in depicting the structure of arterial duct. The larger PDA diameter was suggestive of the lower possibility of spontaneous close.

Objective To investigate the anti-apoptosis effect of transgiutaminase 2 (TG2)in osteosarcoma cell line MG-63 and to explore the mechanism of inhibiting apoptosis of tumor cells.Methods The TG2-tgrgeted siRNA was designed,and the hypoxia culture model of MG-63 cells was established by a hypoxia incubator and the cells were divided into four groups:normal oxygen group,the cells were cultured under normal oxygen;hypoxia group, the cells were cultured in hypoxic incubators;control siRNA hypoxia group,the cells were cultured in hypoxic incubators after transfection of control siRNA;TG2 siRNA hypoxia group , the cells were cultured in hypoxic incubators after transfection of TG2 siRNA.The expressions of Bax and cytochrome C (Cyt C)and the apoptotic rates were observed at different time (6,12,24,48,and 72 h)after hypoxia culture.Microtiter plate assay was performed to monitor the intracellular TG2 activity.RT-PCR was used to detect the mRNA expressions of TG2 and Bax.The expressions levels of TG2,Bax and Cyt C were observed by immunohistochemical staining and Western blotting method.The apoptotic rates were analyzed using flow cytometry.Results Compared with normal oxygen group,the activity of TG2,the mRNA and protein expression levels of TG2 in hypoxia group and control siRNA hypoxia group were increased remarkably with the prolongation of the hypoxia time (P<0.01);the expression level of Bax protein was decreased significantly (P<0.01 ), but the expression level of Bax mRNA had no significant change (P>0.05);the expression levels of Cyt C protein in cytoplasm and mitochondria and the apoptotic rates had no markedly changes (P>0.05).Compared with hypoxia group and control siRNA hypoxia group,the expression levels of mRNA and protein of TG2 in TG2 siRNA hypoxia group were decreased significantly at different time points (P<0.01);the protein expression levels of Bax and Cyt C in cytoplasm and the apoptotic rates were increased markedly (P<0.01 );the expression level of Cyt C in mitochondria was decreased (P<0.01).Conclusion TG2 can inhibit the apoptosis of tumor cells through down-regulating the Bax expression and preventing Cyt C releasing into the cytoplasm.

[Objective]This study was aimed to evaluate treatment outcomes and toxicity of continuous-infusion EPOCH regimen for NK/T-cell lymphoma(NK/TCL).[Methods]From June 2003 to June 2008,34 patients including 30 nasal NK/TCL (88.2%)and 4 nasal type NK/TCL(11.8%)received doxorubicin,vincfistine,etoposide over 96 hours infusion with bolus eyelophosphamide and oral predinisone(EPOCH)chemotherapy as first-line treatment.Median cycles of EPOCH administered were 2.5(1-6 cycles).Additional involved field radiation therapy(IFRT)was administered to patients with localized nasal focus after chemotherapy.[Results]Among 34 patients,33 were eligible for response evaluation.The response rate(RR)was 60.6% (20/33)with complete remission(CR)rate of 45.5%(15/33).The RR of patients with nasal NK/TCL was 66.7%(20/30)with CR rate of 50%(15/30).Only one of the 3 nasal type NK/TCL patients achieved stable disease(SD),the other 2 had progressive disease(PD)during chemotherapy.After a median follow-up of 22(2-68)months,the estimated 3-year overall survival rate(OS)was 52.2%.For patients with nasal NK/TCL,the estimated median survival time was not reached,the 3-year OS was 59.4%.For patients with nasal type NK/TCL,the estimated median survival time was only 7 months.The CR rate was 75.0% for localized nasal NK/TCL who received initial EPOCH chemotherapy followed IFRT with the 3-year OS of 75.0%.Major adverse effect was myelosuppression.The incidence of grade Ⅲ～Ⅳ neutropenia was 30.9%.No treatment-related mortality occurred.[Conclusions]EPOCH regiment was effective and well tolerant for nasal NK/TCL.Combined EPOCH chemotherapy followed by IFRT produced promising outcome for patients with localized disease.However,patients with nasal type NK/TCL responded poorly and more efficacious treatment strategies are urgently needed.

Objective To build the indicators system for clinical pathway management as required by clinical pathway control.Methods An indicators system was proposed by means of evidence-based review,focus group discussions,and ratings of the indicators' importance by doctors and nurses.A multidisciplinary panel of 60 experts from across the country were selected.A 3-round Delphi survey was made on the proposed indicators.The weights of the indicators were established by analytical hierarchy process (AHP).The response rate,Cronbach's α,and the authority coefficient of experts were used as a measure of reliability.Results The response rates of the 3 rounds were 85％,70％,and 94％; the experts authority coefficient was 0.80.The ccoefficient of variation falls with the rising number of consultations.The Kendall's W ranged from 0.40 to 0.83.Following the 3 rounds,consensus was achieved among experts as such a system comprising three first-level,9 second-level,and 36 third-level indicators.Conclusion The expert consultation has achieved reliable results.The established indicators system can serve as a useful instrument for standardized development of clinical pathways management and constant improvement.

China ; Humans ; Logistic Models ; Non-alcoholic Fatty Liver Disease ; Risk Factors

OBJECTIVE:To improve the system of management and maintenance for the purification air-conditioning system in PIVAS,and to further strengthen the management of cleaning environment. METHODS:The cleanness monitoring project of purifi-cation air-conditioning system in PIVAS of our hospital was introduced in terms of temperature and humidity record,pressure differ-ence record,airborne particles detection,settling microbe monitoring report. And the monitoring results were analyzed. RESULTS:The temperature and humidity,pressure difference of clean area in PIVAS of our hospital are both in line with the standard of Phar-macy Intravenous Admixture Quality Management Specification (2010 edition),i.e. temperature at 18-26 ℃,relative humidity of 40%-65%;negative pressure difference between antibiotics,hazardous drug dispensing area and second dressing room are 5-10 Pa. The number of airborne particles (average static particle/m3) at various cleanness degrees in clean area are all in line with the standard of GMP(2010 edition),i.e. maximal allowable number of airborne particles(≥0.5 μm)were 3 520/m3(100 degree);352 000/m3 (10 000 degree);3 520 000/m3 (100 000 degree). The percentage of qualified static settling microbe detection reach 100%in clean area,which is in line with the standard of Settling Microbe Detection Method in Clean Room(Area) of Pharmaceu-tical Industry,i.e. criteria for settling microbe(90 mm)CFU/0.5 h≤1(100 degree);≤3(10 000 degree);≤10(100 000 degree). The percentage of qualified dynamic settling microbe detection is in low level,especially those of dispensing room and secondary dressing room only reaches 80%. CONCLUSIONS:It’s important for effective hospital infection control in PIVAS,the quality im-provement of intravenous injection,the safety guarantee of drug use in patients to further improve standard operation procedure of purification air-conditioning system management and maintenance,and manage and maintain the purification air-conditioning sys-tem completely and scientifically.

OBJECTIVE:To establish a method for content determination of schizantherin E,gomisin J,angeloylgomisin H,schisantherin A,schisantherin B,schisanhenol,anwuligan,schizandrin A,schizandrin B and schizandrin C in Wuzhi tablets.METHODS:RP-HPLC method was adopted.The determination was performed on Symmetry C18 column with acetonitrile-0.1％ phosphoric acid solution (gradient elution) at the flow rate of 1.0 mL/min.The detection wavelength was set 225 rm,and the column temperature was 30 ℃.The sample size was 10 μL.RESULTS:The linear ranges of schizantherin E,gomisin J,angeloylgomisin H,schisantherin A,schisantherin B,schisanhenol,anwuligan,schizandrin A,schizandrin B and schizandrin C were 2.25-67.5ng(r=0.999 6),2.1-63 ng(r=0.999 8),28-840 ng(r=0.999 9),124.6-3 738 ng(r=0.999 9),22.7-681 ng(r=0.999 9),32.7-981 ng(r=0.999 9),47-1 410 ng(r=0.999 9),208-6 240 ng(r=0.999 9),5.36-160.8 rig(r=0.999 9),4.48-134.4 ng(r=0.999 8).The limits of quantitation were 14.17,13.32,9.33,11.37,14.62,19.88,14.66,12.50,16.40,13.55 rg.The limits of detection were 4.62,4.60,3.08,3.76,4.81,6.74,4.93,4.16,5.86,5.03 ng.RSDs of precision,stability and reproducibility tests were less than 3.0％;the recoveries were 96.36％-100.00％(RSD=1.83％,n=6),95.00％-100.00％(RSD=2.07％,n=6),95.00％-98.00％(RSD=1.22％,n=6),95.37％-98.91％ (RSD=1.29％,n=6),95.62 ％-103.71％ (RSD=2.85％,n=6),97.33％-102.67％ (RSD=2.00％,n=6),95.00％-99.33％ (RSD=1.75％,n=6),97.24％-104.93％ (RSD=2.63％,n=6),95.00％-97.50％ (RSD=1.42％,n=6),96.00％-102.00％ (RSD=2.45％,n=6),respectively.CONCLUSIONS:The developed method is accurate,sensitive and reproducible,and it can be used for content determination of 10 lignanoids in Wuzhi tablets.

Background Stromal-derived factor 1α /chemokine receptor 4(SDF-1α/CXCR4) axis is one of the important signals which mediates several different activities such as chemotaxis,adhesion,proliferation and survival resulting in recruitment to sites of immune and inflammatory reactions.Considerable evidence suggests that CXCR4/SDF-1α axis is involved in tumor angiogenesis and plays a key role in the development of ocular neovascularization.Objective The purpose of this study was to explore the effect of CXCR4 antagonist on the development of cxperimental corneal neovascularization(CNV).Methods CNV model was established in the left eye of 8-weekold clean BALB/c mouse by putting the filter with 1 mol/L NaOH at the central cornea for 40 seconds.The animals were randomizcd into hyaluronate group and CXCR4 antagonist group,and the edydrops was topically administered respectively on the day of modeling 4 times per day for 14 days.CNV was examined under the slit lamp at the fourteenth day,and then the corneal suspension and section were made.Expressions of CXCR4 mRNA and protein in corneas were detected using RT-PCR and Western blot.The CD31 level in cornea was assayed by flowcytometry and immunochemistry.The expression of VEGF in burned corneas and suspension from mouse peritoneal macrophages stimulated with CXCR4 antagonist in vitro was detected by ELISA.The use of the animal followed Ragulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results Two weeks after corneal alkali burn,the growth of CNV peaked under the slit lamp.Compared with hyaluronate group,CNV was obviously decreased in the CXCR4 antagonist group.Immunochemistry showed that intensity of positive staining for CD31 in cornea in the CXCR4 antagonist group was weaker than the hyaluronate group.Flowcytometry clarified that CD31 positive cells rate was 9.50％ ±2.34％ in the CXCR4 antagonist group and 17.50％ ±3.16％ in the hyaluronate group,showing a significant difference between them (t=-7.312,P＜0.05).In 2,4,7 days after cornea alkali burn,the expressions of CXCR4 mRNA and protein were significantly enhanced in burn corneas compared with normal corneas(P＜0.01 ;P＜0.05).ELISA showed that the VEGF expression level in corneal tissue and supernatant of mouse peritoneal macrophages in vitro were significantly lower in the CXCR4 antagonist group than that of hyaluronate group(t =10.927,5.151,P＜0.05).The expression level of VEGF in corneal suspension was lower in the GM-CSH+CXCR4 antogonist group than that in the GM-CSH group (P＜0.05).Conclusions CXCR4 antagonist can reduce experimental CNV by down-regulating VEGF expression in cornea.