1.Effects of static pressure on the cytocompatibility of adipose-derived stem cells on electrospun nanofibers
Liang CHEN ; Yi BAI ; Kai LIU ; Wei ZHANG
Chinese Journal of Tissue Engineering Research 2016;20(30):4463-4468
BACKGROUND:Electrospun polylactic acid/polycaprolactone nanofibers (ENF) are a kind of self-synthesized biodegradable material. Our preliminary studies have indicated that the biomaterial exhibits excel ent biocompatibility;however, the research about its mechanics is stil little. OBJECTIVE:To explore the effects of static pressure on the cytocompatibility of adipose-derived stem cel s on the ENF scaffold. METHODS:Adipose-derived stem cel s were seeded onto the ENF scaffold, and then cultured in the low-glucose DMEM supplemented with 10%fetal bovine serum. The mixed constructs were submitted to the static pressure at 0, 15, 30, and 45 kPa for 4 hours using a static pressure device, respectively. Subsequently, the proliferation, adhesion and viability of adipose-derived stem cel s on the ENF scaffold were detected using MTT assay and living/dead staining to evaluate the cytocompatibility. RESULTS AND CONCLUSION:MTT assay showed that there were significant differences in absorbance values among groups by one-way analysis of variance after 4 hours of loading with different static pressures in vitro. Under 0-30 kPa static pressure, the absorbance values increased with static pressure, but the absorbance values declined until the pressure reached 45 kPa, and multiple comparisons between groups showed significant difference. The significant differences in the cel attachment percentage by MTT assay could be found among groups. The living/dead staining results supported the above findings. Furthermore, the significant differences in percentage of living cel s among groups were shown using either one-way analysis of variance or paired t test. In conclusion, the appropriate static pressure can promote the cytocompatibility, proliferation, adhesion and viability of adipose-derived stem cel s on the ENF scaffold. But the excessive pressure is likely to inhibit the cel ular biological behaviors, thus affecting cytocompatibility of adipose-derived stem cel s with the ENF scaffold.
2.Risk factors for positive surgical margins in extraperitoneal laparoscopic radical prostatectomy
Kai HONG ; Lulin MA ; Yi HUANG ; Guoliang WANG ; Lei LIU
Chinese Journal of Urology 2008;29(4):271-274
Objective To determine relative risk factors for positive surgical margins in extraperitoneal laparoscopic radical prostatectomy(LRP). Methods From February 2004 to September 2007,33 patients(mean age 70 years old)with prostate cancers underwent extraperitoneal LRP.All patients were diagnosed by pathology preoperatively.Gleason score:3+3 14 cases(43%),3+4 11 cases(33%),4+3 6 cases(18%),4+4 2 cases(6%).Clinical stage:T1a-T1b 4 cases(12%),T1c 14 cases(43%),T2a-T2b 5 cases(15%),T2c10 cases(30%).Logistic regression analyses were performed. Results LRP was successfully performed on 31 cases.There were 2 cases converted to open surgery.Nine cases(27%)had PSMs.There were 6 cases(67%)and 4 cases(17%)of clinical stage T2c in PSM and negative surgical margin(NSM)groups respectively(P=0.010).There were 3 cases(33%)and 0(0)with high Gleason score(higher than 7)in PSM and NSM cases(P=0.015).There were 4 cases(44%)and 5 cases(21%)with t-PSA higher than 20dg/ml in PSM and NSM cases respectively(P=0.178).In these 9 cases,there were 4 cases(44%)positive with DRE.However there were 9 in the 24 NSM cases(38%)(P=0.509).Clinical stage T2c was independently positively correlated with PSM(OR=24.69).High Gleason score(higher than 7)and t-PSA higher than 20 ng/ml were positively correlated with PSM. Conclusions Clinical stage is positively correlated with PSM.It is an independent factor.High Gleason score(higher than 7)and t-PSA higher than 20 ng/ml mignt be the risk factors in predicting PSM and should be used together with clinical stage.Positive DRE findings may be also useful to predict PSM.
3.Relation between drug release and the drug status within curcumin-loaded microsphere.
De CHEN ; Yi LIU ; Kai-yan FAN ; Yi-qiao XIE ; An-an YU ; Zi-hua XIA ; Fan YANG
Acta Pharmaceutica Sinica 2016;51(1):140-146
To study the relation between drug release and the drug status within curcumin-loaded microsphere, SPG (shirasu porous glass) membrane emulsification was used to prepare the curcumin-PLGA (polylactic-co-glycolic acid) microspheres with three levels of drug loading respectively, and the in vitro release was studied with high-performance liquid chromatography (HPLC). The morphology of microspheres was observed with scanning electron microscopy (SEM), and the drug status was studied with X-ray diffraction (XRD), differential scanning calorimetry (DSC) and infrared analysis (IR). The drug loading of microspheres was (5.85 ± 0.21)%, (11.71 ± 0.39)%, (15.41 ± 0.40)%, respectively. No chemical connection was found between curcumin and PLGA. According to the results of XRD, curcumin dispersed in PLGA as amorphous form within the microspheres of the lowest drug loading, while (2.12 ± 0.64)% and (5.66 ± 0.07)% curcumin crystals was detected in the other two kinds of microspheres, respectively, indicating that the drug status was different within three kinds of microspheres. In the data analysis, we found that PLGA had a limited capacity of dissolving curcumin. When the drug loading exceeded the limit, the excess curcumin would exist in the form of crystals in microspheres independently. Meanwhile, this factor contributes to the difference in drug release behavior of the three groups of microspheres.
Calorimetry, Differential Scanning
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Curcumin
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chemistry
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Drug Liberation
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Lactic Acid
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Microscopy, Electron, Scanning
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Microspheres
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Polyglycolic Acid
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X-Ray Diffraction
4.Modified limited L incision with distraction bone block arthrodesis for subtalar osteoarthritis.
Yi LI ; Hong-Mou ZHAO ; Xiao-Jun LIANG ; Cheng LIU ; Kai ZHAO ; Jie YANG
China Journal of Orthopaedics and Traumatology 2014;27(7):536-539
OBJECTIVETo evaluate the functional outcomes of modified limited "L" incision beside the Achilles tendon with distraction bone block arthrodesis in treatment of subtalar osteoarthritis.
METHODSFrom March 2009 to September 2012, a total of 22 cases of old calcaneus fractures with subtalar osteoarthritis were treated with modified limited "L" incision and distraction bone block arthrodesis including 13 males and 9 females with a mean age of 35.3 years old (ranged 22 to 49). The mean time from calcaneal fracture was 21 months (ranged 11 to 32). According to the Stephens-Sanders classification, 16 cases were type II and 6 were type III. The modified-AOFAS ankle-hindfoot score was used for functional outcomes evaluation.
RESULTSThere was one incision necrosis and no infection, implant failure, bone-graft absorbed or talus necrosis was note at the follow-up time. A total of 21 cases were followed up for a mean time of 29 months (ranged from 18 to 46 months). All of the cases reached a bony union within 4 months postoperation. The mean modified-AOFAS ankle-hindfoot score was 82.6 points (ranged from 66 to 92 points),reached a significantly improvement in comparing with the mean preoperative score (50.8 points,ranged from 32 to 65 points, P < 0.01).
CONCLUSIONThe modified limited"L" incision beside the Achilles tendon with distraction bone block arthrodesis is an acceptable and alternative treatment method for subtalar osteoarthritis. This method is easy to use and with less complication. It can correct the main pathological changes and reach good functional outcomes.
Adult ; Arthrodesis ; methods ; Female ; Humans ; Male ; Middle Aged ; Osteoarthritis ; surgery ; Subtalar Joint ; surgery
5.Influence of different immunosuppressants on monocyte chemoattractant protein-1 secretions in the whole blood
Ming WANG ; Yi HE ; Zhanguo LIU ; Yuwei LUO ; Kai WU ; Erwei SUN ; Ming ZHAO
Chinese Journal of Tissue Engineering Research 2010;14(18):3314-3317
BACKGROUND: In the field of organ transplantation, patients often take immunosuppressants after organ transplantation, such as CsA, FK506, DEX and MPA. However, their mechanisms of immunosuppression are different. The effect of immunosuppressive drugs on monocyte chemoattractant protein-1 (MCP-1) remains poorly understood. OBJECTIVE: To investigate the effects of different immunosuppressants on the secretions of MCP-1 in whole blood. METHODS: The whole blood of healthy volunteers was mixed with different immunosuppressants for 6 hours, such as CsA, FK506, DEX and MPA, which included low, middle and high concentrations, followed by PMA and IONO stimulation for 6 hours. MCP-1 levels in whole blood samples were compared. The whole blood cultured alone served as control. RESULTS AND CONCLUSION: MCP-1 secretion was inhibited by DEX (1, 10 mg/L) and CsA (0.25,1.25 mg/L)- However, FK and MPA exhibited no such effect. Therefore, DEX and CsA may inhibit the function of monocytes and macrophages in immune system by diminishing the secretion of MCP-1. The combination of FK (5 μg/L), MPA (10 mg/L) and DEX (1 mg/L) or CsA (0.25 mg/L), MPA (10 mg/L) and DEX (1 mg/L) can inhibit the secretion of MCP-1, but only DEX among all the immunosuppressants mentioned above exhibited significant effect on inhibiting the secretion of MCP-1 when using alone.
6.Knockdown of EphB4 gene by small interference RNA inhibits proliferation of glioma cells in vitro
Chunhui LI ; Zhihong LI ; Yanfang SHI ; Yi GUO ; Yanan DING ; Haipeng LIU ; Kai CUI
Chinese Journal of Nervous and Mental Diseases 2010;36(3):129-132
Objective To explore the role of EphB4 in proliferation of glioma cells. Methods The mRNA and protein expressions of EphB4 were detected using RT-PCR, immunochemistry, and Western-blot, respectively. EphB4 siRNA was synthesized and transfected into U251 cells using Lipofectamine 2000. Glioma cell proliferation, apoptosis, and invasion were determined by MTT assay, TUNEL and transwell experiment, respectively. Results The expression (P<0.05) and proliferation of EphB4 were obviously decreased in U251 transfected with EphB4 siRNA and the proliferation was further decreased with the increased concetrations of siRNA. Compared with U251 group and siRNASCR group, EphB4 siRNA at different concentrations (25, 50 or 100 nmol/L) significantly reduced the invasion ability of cells and increased the number of apoptotic cells (P<0.05). Conclusions EphB4 plays an important role in the regulation of glioma cell proliferation, apoptosis and invasion.
7.Distribution and prognostic significance of estrogen receptors expression in prostate cancer
Yuqing LIU ; Jian LU ; Min LU ; Ran MA ; Kai HONG ; Yi HUANG ; Lulin MA
Chinese Journal of Urology 2013;(5):378-383
Objective To evaluate the expressions of estrogen receptor (ER) α and β in human prostate cancer and adjacent non-cancerous tissues,and to evaluate the correlation between the expression and the clinicopathological features and prognosis.Methods Immunohistochemical staining was used to detect ERα and ERβ in 85 prostate adenocarcinoma tissues,adjacent non-cancerous tissues,and 29 benign prostatic hyperplasia (BPH) tissues.The correlation between the expression and the clinicopathological features was analyzed by Spearman's coefficient.Cox's proportional hazards regression model was used to identify the risk factors for biological recurrence.Results There were significant differences between the expression of ERα in prostate cancer,adjacent non-cancerous tissues,and BPH [epithelial cell 0 (0/85),11.8% (10/85) and 24.1% (7/29),P=0.000; and in stromal cell 52.9% (45/85),67.1% (57/85),31.0% (9/29),P =0.003].There were significant differences between the expression of ERβ in these groups [epithelial cell 36.5% (31/85),61.2% (52/85),100.0% (29/29),P =0.000; and in stromal cell 49.4% (42/85),72.9% (62/85),79.3% (23/29),P =0.001].The ERα expression in cancerous stromal cells was positively correlated with the PSA level (r =0.296,P =0.006) and Gleason score (r =0.404,P =0.000).The ERβ expression was negatively correlated with Gleason score in cancerous epithelial cells (r =-0.254,P =0.019) and stromal cells (r =-0.315,P =0.003).Multivariate analysis revealed that negative expression was an independent poor prognostic factor for the biological recurrence free survival after radical prostatectomy (HR =0.107,95.0% CI 0.019-0.592,P =0.010).Conclusions There were significant differences between the expression of ERα and ERβ in prostate cancer,adjacent non-cancerous tissues,and BPH.ERα in cancerous stromal cells,ERβ in cancerous epithelial and stromal cells were related to the differentiation of prostate adenocarcinoma.ERβ in cancerous epithelial cells can be used as an independent prognostic factor for biological recurrence after radical surgery.
8.Effect and impact of holmium laser versus thulium laser enucleation of the prostate on erectile function.
Kai HONG ; Yu-qing LIU ; Jian LU ; Chun-lei XIAO ; Yi HUANG ; Lu-lin MA
National Journal of Andrology 2015;21(3):245-250
OBJECTIVETo compare the effect and impact of holmium laser enucleation of the prostate (HoLEP) and 120-W thulium: YAG vapoenucleation of the prostate (ThuVEP) on erectile function in the treatment of benign prostatic hyperplasia (BPH).
METHODSWe retrospectively analyzed 93 cases of symptomatic BPH treated by HoLEP or 120 W ThuVEP. We made comparisons between the two groups of patients in the baseline and postoperative clinical and surgical indexes as well as their IPSS, quality of life (QOL), maximum flow rate (Qmax), postvoid residual urine volume (PVR), and IIEF-EF scores before surgery and during the 12-month follow-up.
RESULTSThuVEP, in comparison with HoLEP, achieved a significantly shorter operation time ([57.6 +/- 12. 8] vs. [70.4 +/- 21.8] min, P = 0.001) and a higher laser efficiency ([0.71 +/- 0.18] vs. [0.62 +/- 0.19] g/min, P = 0. 021). At 1, 6, or 12 months of follow-up, no significant differences were observed in IPSS, OOL, Omax, and PVR between the two groups (P > 0.05). Both the HoLEP and ThuVEP groups showed low incidences of complications and remarkably improved IIEF-EF scores at 12 months postoperatively, but with no significant differences (both P > 0.05). However, in those with relatively normal erectile functions before operation, the mean IIEF-EF score was reduced from 22.8 +/- 2.2 preoperatively to 21.0 +/- 2.7 after HoLEP, (P = 0.036).
CONCLUSIONBoth HoLEP and 120W ThuVEP are effective and safe in the treatment of BPH. Compared with HoLEP, 120 W ThuVEP has even a higher laser efficiency. However, neither can significantly improve erectile function, and HoLEP may have a short-term negative impact on the relatively normal erectile function of the patient.
Aged ; Holmium ; Humans ; Laser Therapy ; adverse effects ; methods ; Lasers, Solid-State ; therapeutic use ; Male ; Middle Aged ; Penile Erection ; Prostatectomy ; adverse effects ; methods ; Prostatic Hyperplasia ; surgery ; Quality of Life ; Retrospective Studies ; Thulium ; Transurethral Resection of Prostate ; Treatment Outcome ; Urine
9.Study on Quick Identification of Fufang Yuxingcao Tablet by Near-infrared Spectroscopy
Yi LU ; Yan YAN ; Kai YAN ; Kaishuang LIU ; Guo YIN ; Jue WANG ; Tiejie WANG
China Pharmacy 2016;27(24):3420-3421,3422
OBJECTIVE:To establish a method of quick identification of Fufang yuxingcao tablel(manufactured by A). METH-ODS:NIRDRS was adopted,the near-infrared diffuse reflectance spectrum were recorded by a fiber optic probe,and the conformi-ty test model was established through the consistency index (CI) value and the CI limit comparison method. The spectral range of 9 002-7 497.8 cm-1 ,6 903.8-5 596.4 cm-1 and 5 002.4-4 246.4 cm-1 was selected as the characteristic spectral band ,the spectra were preprocessed by first derivation and vector normalization with 17 smoothing points , and 7.0 was set as the CI value;the characteristic spectrum correlation coefficient model was established through the correlation coefficient method ,the spectral range of 6 200-5 500 cm-1,5 000-4 700 cm-1 was selected as the characteristic spectral band,the spectra were preprocessed by first deri-vation with 17 smoothing points,and 97% was set as the threshold. RESULTS:The above established models can rapidly identify and accurately distinguish Fufang yuxingcao tablet(manufactured by A)from similar products of other manufacturers,the CI value of the validation samples was beyond 7.0,the correlation coefficient(r)were less than 97% compared with the reference sample. CONCLUSIONS:The method is simple and rapid,and can be used for fast screening of Fufang yuxingcao tablet.
10.Construction of eukaryotic vector of monkey B virus glycoprotein D gene and the gD gene expression
Xin WANG ; Simeng YI ; Huifang LIU ; Kai MA ; Junwen FAN ; Yunan MA ; Ying YOU ; Zhaozeng SUN
Chinese Journal of Comparative Medicine 2015;(6):28-31,81
Objective To establish an eukaryotic vector of monkey B virus glycoprotein D gene and analyze the expression of gD gene in human embryonic kidney 293T cells.Method First, the protein of monkey B virus glycoprotein D was obtained by gene synthesis.The gene fragments were digested with Pst I and Not I, and ligated to pEGPF-N3. Then, the recombinant plasmid pEGPF-N3-GD was transfected into 293T cells.The expression of gD protein in the cells was detected by Western blot, and the expression localization was investigated using laser scanning confocal microscopy. Results The recombinant plasmid pEGPF-N3 carrying gD gene was successfully constructed, and normally expressed in the 293T cells.Conclusions Glycoprotein D of monkey B virus is expressed successfully in the 293T cells and the protein is located on the cell surface.It may be useful for the preparation of specific recombinant antigen to the glycoprotein D of monkey B virus on cell surface, and can be also used for preparation of antigen slide for detection of monkey B virus.