Tissue engineering provides urologists a new way to fix or reconstruct the impaired organs.Reconstitution of corporal bodies of penis with engineered tissue substitutes has been applied in animal models.In hypospadias reconstruction,the use of engineered tissue substitutes has been applied clinically.The clinical application of bladder tissue substitutes has been ongoing phase II clinical trial.Great progress has been made in renal replacement therapy with clinical application of human progenitor cells in hemofiltration units,and the engineered intracorporeal renal replacement unit will come true by additional studies.The current status of tissue engineering in clinical practice of urology is reviewed in this paper.

Objective To investigate the effects of anti-cell membrane associated DNA (mDNA) antibodies in the diagnosis of systemic lupus erythematosus (SLE) lacking of specific autoantibodies including anti Sm,anti ds-DNA,and anti-nucleosome antibodies.Methods Indirect immunofluorescence assay was used to measure anti-mDNA antibodies in serum of 145 SLE patients,and indirect immunofluorescence,Western-blot and ELISA were used to detect the anti-dsDNA ,anti-Sm and anti- nueleosome antibodies respectively to analysis the value of anti-mDNA antibodies on the specific autoantibodies negative patients with SLE.Results The sensitivity for anti-mDNA antibodies (69.7%) in SLE was significantly higher than anti-Sm (19.7%),anti-dsDNA ( 31.9% ) and anti-nucleosome (45.8% ).The incidences of anti-mDNA antibodies in SLE lacking of anti-dsDNA,Sm and anti-nueleosome antibodies (AnuA) were 64.3% ,70.2% and 60.3% respectively.Conclusion Anti-mDNA antibodies are serologic marker of SLE and important in diagnosis of SLE lacking of anti-dsDNA,Sm and nucleosome antibodies.

Objective To study the effect of Xin Kang Injection on cardiac function of rats with adriamycin-induced myocarditis and to explore its pharmacological mechanism.Methods Seventy Wistar rats were randomly divided into seven groups:normal control group,model group,three-dosage Xin Kang Injection groups(high-,moderate-and low-dosage respectively),deslanoside group,and Huangqi group.The myocarditis rat model was established by intraperitoneal injection of adriamycin.Left ventricular end-diastolic pressure(LVEDP),left ventricular systolic pressure(LVSP),heart rate(HR),T value,and ? DP/DTmax were observed.Results LVSP,+ DP/DTmax in the treatment groups were higher and-DP/DTmax lower than those in the model group(P

Objective To evaluate the feasibility of apply Ion Proton semiconductor sequencing platform in non-invasive prenatal genetic diagnosis .Methods Totally 1 000 pregnant women with a singleton pregnancy of 12-32 weeks gestation were selected from the Third affiliated Hospital of Zhengzhou University from Jan to Dec 2013.Using noninvasive prenatal genetic diagnosis based on Ion Proton semiconductor sequencing platform to study their cffDNA .In parallel, 72 pregnant women received invasive prenatal diagnosis by traditional chromosomal analysis with amniocentesis chorionic villus sampling .Results It′s shown that 18 out of 1 000 (1.8%) pregnant women underwent the noninvasive prenatal genetic testing had a high risk for aneuploid chromosomes , including 7 cases of 21-trisomy, 4 cases of 18-trisomy, 2 cases of 13-trisomy, 4 cases of sex chromosomal abnormality , and 1 case of 15-trisomy.It demonstrated that the rate and accuracy of fetal 21-trisomy, 13-trisomy and 18-trisomy by non-invasive prenatal genetic testing were both 100%without misdiagnosis , the rate of detection for sex chromosomal abnormality was 2/2 with a false positive rate of 1/3.However, the 15-trisomy predicted by the non-invasive prenatal diagnosis in a woman was finally proved to be a false positive .Based on the results by karyotyping (55/55) as well as follow-ups (493/493), the specificity of the non-invasive prenatal diagnosis for detection of 21-trisomy, 18-trisomy and 13-trisomy was 100%.One Ion PITM chip could detect 12 to 15 samples in 1.5 h and the whole process of noninvasive detection could be completed in 1 to 1.5 days.Conclusions The non-invasive prenatal diagnosis by Ion Proton semiconductor sequencing platform could provide fast and accurate detection of fetal aneuploidy .The benchtop high-throughput sequencing platform has laid the foundation for the independent application in clinical settings for fetal aneuploidy detection .

Objective To dig the correlation between genes and four TCM constitutions (phlegm-dampness, blood stasis, allergy, and dampness-heat); To analyze microscopic mechanism of constitution formed and constitution-disease correlation; To provide biological basis for TCM constitution classification and constitution-disease correlation theory. Methods Literature about TCM constitutions and gene expressions in CNKI and Wanfang Database were retrieved by computers from the establishment of database to 31, December 2016. Differentially expressed genes of four TCM constitutions were collected, and DAVID database was used for enrichment analysis, including gene tissue enrichment analysis, disease analysis, GO analysis, and pathway analysis. The results of DAVID database analysis were downloaded and analyzed. Results Four TCM constitutions had differences in gene level, and each constitution had the specificity in significantly enriched gene function, activity, pathways, related diseases, and expression locations. Different patterns of gene expression led to the constitution differences in disease susceptibility and tendency. Conclusion The essence of TCM constitution formation is the change of gene expression patterns, and the pathological changes are also caused by the change.

Aim To investigate the effect of curcumin extract on melanin production and melanosome transport, and to explore the possible mechanism of the curcumin extract on microenvironment. Methods (1) B16F10 and HaCaT cells were cultured with different concentrations of curcumin. The proliferation ability was detected by MTT method. Melanin synthesis and tyrosinase activity in B16F10 cells were detected by NaOH pyrolysis method and Oxidation dopamine response in vitro. The expression levels of key proteins were detected by Western blot. (2) B16F10 cells were cultured with different concentrations of ISG15 protein. NaOH pyrolysis method and Oxidation dopamine response in vitro were used to detect melanin synthesis and tyrosinase activity in B16F10 cells. Results Curcumin could directly inhibited tyrosinase activity and melanin production, and inhibit melanocyte migration within a certain concentration range. ISG15 protein could enhance the melanin production, tyrosinase activity. Curcumin could reduce the expression of the ISG15 in HaCaT cell, change the microenvironment of melanocyte, and indirectly inhibit melanin synthesis through ISG15. Conclusions In addition to directly inhibiting melanin synthesis, curcumin can also play an indirect role in inhibiting melanin synthesis by inhibiting the expression of ISG15 protein and altering the microenvironment of melanocytes.

OBJECTIVETo explore the feasibility of creating a 3D-CAD model of craniofacial prostheses through mirror technique to repair the unilateral craniofacial defects and restore craniofacial symmetry.

METHODSPatients with unilateral craniofacial defects underwent spiral CT scanning. CAD3-D image was reconstructed ad 3-D CAD model of craniofacial prosthesis was created with mirror technique, Boolean operation and rapid prototyping technique. Then the prosthesis made of bioactive artificial bone was made through plaster cavity block.

RESULTS15 cases were treated with no complications. Good symmetry was achieved after operation.

CONCLUSIONSDesigning the craniofacial prosthesis with mirror technique guarantees excellent functional and cosmetic results for repairing the unilateral craniofacial defects.

Adolescent ; Adult ; Child ; Computer-Aided Design ; Female ; Humans ; Imaging, Three-Dimensional ; Male ; Middle Aged ; Prosthesis Design ; methods ; Skull ; diagnostic imaging ; pathology ; surgery ; Tomography, Spiral Computed ; Young Adult

OBJECTIVETo investigate the expression of BRCA1 in esophageal squamous cell carcinoma (ESCC) tissues and evaluate its correlation with clinicopathological features as well as the prognosis of ESCC patients.

METHODSThe expression of BRCA1 was detected by immunohistochemistry (IHC) in 201 specimens of T3 stage ESCC tissues and corresponding adjacent normal tissues using tissue microarray. The correlation between BRCA1 expression and clinicopathological features of ESCC was determined by chi-square analysis. The cumulative survival rate was analyzed by Kaplan-Meier method.

RESULTSThe positive rate of BRCA1 expression in ESCC tissues was significantly higher than that in adjacent normal tissues [88.6% (178/201) vs. 36.8% (74/201), P < 0.001]. There was a significant correlation between the expression of BRCA1 and lymph node metastasis. In the tumors with positive lymph nodes, strong positive expression of BRCA1 was found in 45.0% (49/109), while only 19.6% (18/92) in tumors without lymph node metastasis, showing a significant difference (P < 0.001). A close relationship was also found between the expression of BRCA1 and gross typing of tumors (P < 0.05). The expression of BRCA1 was not significantly correlated with gender, age, tumor location, differentiation, and tumor thrombus (P > 0.05). The results of Kaplan-Meier analysis indicated that ESCC patients with a higher positive rate of BRCA1 expression have a poorer prognosis (P < 0.05).

CONCLUSIONSThe expression of BRCA1 is related to the occurrence and development of esophageal carcinoma. BRCA1 protein may serve as a new potential biomarker in estimating the biological behavior of ESCC.

Adult ; Aged ; Aged, 80 and over ; BRCA1 Protein ; metabolism ; Biomarkers, Tumor ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; pathology ; surgery ; Esophageal Neoplasms ; metabolism ; pathology ; surgery ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Survival Rate

OBJECTIVETo study the biological effects of the paracrine from ADSC after being stimulated by insulin on vascular endothelial cells.

METHODS(1) ADSC was isolated from human adipose tissue and cultured in vitro. The third generation cells were collected and divided into insulin group (I, cultured with serum-free DMEM containing 1 x 10(-7) mol/L insulin) and control group (C, cultured with serum-free DMEM) according to the random number table, with 6 slots in each group. Three days later, ADSC culture medium (ADSC-CM) was collected for determination of levels of vascular endothelial growth factor (VEGF) and hepatocyte growth factor (HGF) by ELISA. (2) Human umbilical vein endothelial cells (HUVEC) were cultured to the third generation, and they were cultured with special nutrient solution and divided into ADSC-CM with insulin stimulation group (AI), ADSC-CM without insulin stimulation group (AC), insulin group (I, with same concentration as above), blank control group (BC) according to the random number table. Three days later, proliferation of HUVEC was determined with MTT method (with expression of absorbance value). Another two samples of HUVEC were respectively divided into 4 groups as above for determination of apoptosis rate with Annexin V/FITC double-staining 12 hours after culture, and HUVEC migration with scratch adhesion test at post scratch hour (PSH) 12, 24, 36, 48. Data were processed with t test.

RESULTS(1) Compared with those in C group [(287 +/- 47), (577 +/- 84) pg/mL, respectively], the secretion levels of VEGF and HGF in I group [(643 +/- 64), (930 +/- 68) pg/mL, respectively] were significantly increased (with t value respectively 18.869, 18.475, P values all below 0.05). (2) The absorbance value of HUVEC in AI and AC groups was 0.847 +/- 0.042, 0.798 +/- 0.022, respectively, which were higher than that in I and BC groups [0.665 +/- 0.028 (with t value respectively 4.579, 3.732), 0.674 +/- 0.031 (with t value respectively 3.761, 4.073), P values all below 0.01], and that in AI group was higher than that in AC group (t = 2.576, P < 0.05). The apoptosis rates of HUVEC in AI and AC groups [(5.8 +/- 1.9)%, (9.0 +/- 2.0)%, respectively] were obviously lower as compared with that in I and BC groups [(30.4 +/- 6.0)% (with t value respectively 12.891, 10.417), (31.4 +/- 7.4)% (with t value respectively 11.474, 9.783), P values all below 0.05 ], and that in AC group was higher than that in AI group (t = 8.548, P < 0.05). The distance of migration of HUVEC in AI and AC groups were greater than that in I and BC groups at PSH 36, 48, and that in AI group was greater as compared with that in AC group (with t value respectively 4.076, 4.573, P values all below 0.05).

CONCLUSIONSParacrine from ADSC after being stimulated by insulin can promote proliferation and migration of HUVEC, and suppress its apoptosis, and it is beneficial for tissue vascularization.

Adipocytes ; cytology ; secretion ; Adipose Tissue ; cytology ; Apoptosis ; Cell Movement ; Cell Proliferation ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Hepatocyte Growth Factor ; metabolism ; Human Umbilical Vein Endothelial Cells ; cytology ; metabolism ; Humans ; Insulin ; pharmacology ; Stem Cells ; cytology ; secretion ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo elucidate the effects of interferon-gamma (IFN-gamma) on the transforming growth factor beta (TGF-beta)/Smad pathway in keloid-derived fibroblasts (KFb), and to investigate the underlying mechanism in the treatment of pathologic scar with IFN-gamma.

METHODSKeloid tissue of 3 patients were obtained, and then KFb were separated and cultured in vitro. KFb from passages 3 to 5 were used for the study. (1) KFb were divided into control group (incubated with serum-free DMEM), TGF-beta(1) group (treated with 10 ng/mL TGF-beta(1)), IFN-gamma group (treated with 100 ng/mL IFN-gamma), and TGF-beta(1)+IFN-gamma group (incubated with 10 ng/mL TGF-beta(1) combined with 100 ng/mL IFN-gamma). The expression level of mRNA and protein of connective tissue growth factor (CTGF), alpha smooth muscle actin (alpha-SMA) protein and expression of alpha-SMA positive KFb were detected by real-time fluorescent quantitation RT-PCR (FQ-RT-PCR), Western blot and immunofluorescence cytochemical staining. (2) Another sample of KFb was obtained and treated with 10 ng/mL IFN-gamma. The expression level of Smad 3 and Smad 7 protein was detected by Western blot before and 1, 2, 4, 6, 8 h post stimulation (PSH). The expression level of Smad 3 and Smad 7 mRNA was assessed by FQ-RT-PCR before stimulation and 30 mins post stimulation and at PSH, 1, 2, 4, 6, 8. (3) Another sample of KFb was obtained and divided into 1, 10 and 100 ng/mL IFN-gamma groups based on the concentration of IFN-gamma, treated for 4 hours; KFb without IFN-gamma treatment was set up as control group. The expression levels of the protein and mRNA of Smad 3 and Smad 7 were measured by FQ-RT-PCR and Western blot.

RESULTS(1) The level of mRNA and protein of CTGF in IFN-gamma group (0.017 +/- 0.009 and 1.198 +/- 0.004) was respectively lower than that in control group (0.024 +/- 0.013 and 1.229 +/- 0.011, P < 0.05). The level of mRNA and protein of CTGF in TGF-beta(1)+IFN-gamma group (0.634 +/- 0.138 and 1.204 +/- 0.010) was respectively lower than that in TGF-beta(1) group (1.331 +/- 0.298 and 1.727 +/- 0.004, P < 0.01). The fluorescence intensity of alpha-SMA positive KFb (0.922 +/- 0.059) and the expression level of alpha-SMA protein (0.3051 +/- 0.0031) in IFN-gamma group decreased significantly than those in control group (1.055 +/- 0.005 and 0.4513 +/- 0.0094, P < 0.01). The fluorescence intensity of alpha-SMA positive KFb (1.129 +/- 0.004) and the expression level of alpha-SMA protein (0.6734 +/- 0.0098) in TGF-beta(1)+IFN-gamma group decreased significantly than those in TGF-beta(1) group (1.270 +/- 0.005 and 1.3842 +/- 0.0024, P < 0.01). (2) The expression level of Smad 3 mRNA and protein at the first time point after IFN-gamma treatment increased temporarily then decreased gradually, and mRNA expression level reached the nadir at PSH 4, it rose gradually later, though it was still lower at PSH 8 than that before treatment (P < 0.01); protein expression level at PSH 8 was significantly lower than that before treatment (P < 0.01). The expression level of Smad 7 mRNA and protein increased gradually to the maximum at PSH 2 and 4 respectively, then decreased but was still higher at PSH 8 than that before treatment (P < 0.05). (3) Compared with those in control group, the expression levels of Smad 3 mRNA and protein in 1, 10 and 100 ng/mL IFN-gamma group were significantly lower, the expression levels of Smad 7 mRNA and protein were significantly higher (P < 0.05 or P < 0.01). The higher concentration of IFN-gamma, the more significant differences were observed.

CONCLUSIONSIFN-gamma can down-regulate the expression of Smad 3 while up-regulate the expression of Smad 7 in a time- and dose-dependent manner, and reduce the expression level of CTGF and alpha-SMA in the basic state or induced by TGF-beta(1), which shows a significant inhibitory effect on the TGF-beta/Smad signal pathway. This may be an important mechanism in the treatment of pathologic scar by IFN-gamma.

Cells, Cultured ; Fibroblasts ; drug effects ; metabolism ; Humans ; Interferon-gamma ; pharmacology ; Keloid ; metabolism ; RNA, Messenger ; genetics ; Signal Transduction ; drug effects ; Smad Proteins ; metabolism ; Transforming Growth Factor beta1 ; metabolism