Three lines of studies (lung in sito, isolated perfused lung and cultured pulmonary artery endothelial cell, PAEC) were undertaken in our laboratory in recent years which demonstrated that oxygen free radicals (OFRs) participates in the pathogenesis of rat lung edema induced by oleic acid (OA). The OFRs derived from xanthine/xanthine oxidase in PAEC might play a primary and initial role, whereas that derived from PMNs might act as an important but secondary and amplifying role in the pathogenesis of lung edema induced by OA.

Objective To investigate the level of CD4+CD25+CD127low regulatory T cells (Tregs) in the peripheral blood of hepatocellular carcinoma (HCC) patients with HBV infection and to explore the relationship between these cells and HBV infection. Methods Peripheral blood mononuclear cells were isolated from 41 HCC patients infected with HBV, 34 HCC patients without HBV infection and 29 healthy persons. These mononuclear cells were labeled with the monoclonal anti-bodies of CD4, CD25 and CD127, and then Tregs level was determined by flow cytometry (FCM). Moreover, the load of HBV DNA in the blood serum of HCC patients with HBV infection was detected by fluorescent quantitative polymerase chain reac-tion (FQ-PCR). Results The percentage of CD4+CD25+CD127low Tregs in CD4+T cells in the HCC patients with HBV infec-tion was higher than that in the HCC patients without HBV infection [(8.24±2.20)%vs (7.06±2.46)%, P＜0.05] and that in the healthy persons [(8.24 ± 2.20)% vs (6.51 ± 1.99)%, P ＜ 0.01]. Furthermore, the percentage of CD4+CD25+CD127low Tregs in CD4+T cells in the HCC patients without HBV infection was higher than that in the healthy persons [(7.06±2.46)%vs (6.51± 1.99)%, P＜0.05]. In addition, the level of CD4+CD25+CD127low Tregs in the HCC patients with HBV infection was gradually increased following the increase of HBV DNA load and there was a positive correlation between the level of CD 4+CD25+CD127low Tregs and the load of HBV DNA (r=0.350,P ＜ 0.05). Conclusion The level of CD4+CD25+CD127low Tregs in HCC patients is increased remarkably and associated with the number of HBV DNA copy, which may be an important mecha-nism of immunologic escape of virus in HCC.

Objective To investigate the expression of stem cell associated protein CD90 in primary hepatocellular carcinoma (HCC) and its relationship with clinical pathological characters ;to analyze the relation between CD90 and epithelial mesenchymal transition (EMT) markers E‐cadherin (E‐cad) and N‐cadherin (N‐cad) .Methods The expressions of CD90 ,E‐cad and N‐cad in 53 tissues of HCC were detected by immunohistochemistry streptavidin‐perosidase (SP ) method , and 10 surrounding of liver benign lesions were studied as control .The expression of E‐cad and N‐cad in CD90 positive cells of two HCC cell lines (M HCC97‐L and HCCLM‐3 ) was measured by flow cytometry .Chi square test and Spearman rank correlation analysis were performed for count data .Independent sample t test was used for measurement data comparison .Results There was no CD90 expression in control liver tissue . The positive rate of CD90 expression in HCC tissue was (34% ,18/53) ,and the difference between two groups was statistically significant (χ2 = 4 .755 , P< 0 .05) .The expression of CD90 in HCC was positively correlated with portal vein cancer embolus (r= 0 .378 , P< 0 .05) and was negatively correlated with histological differentiation degree and capsular infiltration (r= -0 .398 and -0 .519 ,both P<0 .05) .The expression of CD90 was negatively correlated with the expression of E‐cad (r= -0 .341 , P<0 .05) ,however was positively correlated with the expression of N‐cad (r=0 .441 ,P<0 .05) .The positive expression rate of E‐cad in CD90 positive HCCLM‐3 cells was lower than that of MHCC97‐L cells ((2 .56 ± 0 .29)% and (4 .31 ± 0 .18)% ,t= 8 .757 ,P< 0 .05) ,while the positive expression rate of N‐cad was higher than that of MHCC97‐L cells ((8 .10 ± 1 .45)% and (5 .51 ± 0 .44)% ,t= -9 .667 ,P<0 .05) .Conclusions The expression of stem cell associated protein CD90 is correlated with the EMT of HCC .Their interaction promote tumor infiltration metastasis w hich may be a new target of HCC treatment .

Objective To establish the long-term culture system for fetal skin fibroblast by performing long time in vitro cultivation of the cells,and study the potential of its differentiation to hepatocytes.Methods Fibroblast was isolated from human fetus skin tissue.Surface phenotypes of cells were detected by ICC and FCM,and biological characteristics were analyzed by the karyotype analysis and soft agar colony formation observation.ALB、CK18、CK19 were detected by ICC,glycogen stain by PAS,AFP and ALB mRNA by RT-PCR after P3～30cells were induced differentiation by cytokines of HGF,FGF4 and OSM.Results CD29,CD49f,HLA- Ⅰ and CD 105 were highly expressed while CD90 hardly in skin fibroblast.The rate of induced differentiation of fibroblast into hepatocyte-like cells was approximately 5％.The cells could be cultured in vitro for almost 50 passages with normal karyotype and no oncogenic and immortalized characteristics.Conclsion The skin fibroblast possesses the characteristic of mesenchymal stem cell and can be induced into hepatocyte-like cell in vitro.

OBJECTIVE: To investigate biological characteristics of human umbilical cord-derived mesenchymal stem cells, and to explore the possibility of hepatocyte-like cells differentiation.METHODS: The umbilical cord was provided by healthy term birth woman in Tianjin Third Central Hospital. Mesenchymal stem cells were isolated from human umbilical cord by enzyme digestion method. Cells were passaged at 80%-90% confluent. The ninth passage of cells at a density of 5×10~(10)/L were seeded in 12-well culture plate and incubated with DMEM containing hepatocyte growth factor, fibroblast growth factor-4 and oncostatin for 28 days. Cell growth activity was detected by MTT method; cell cycle was detected by flow cytometry; surface immunological marker in MSC was detected by immunocytochemical stain and flow cytometry; specific surface phenotype of hepatocyte was detected by immunocytochemical staining. Function characteristic of hepatocyte was determined by staining for glycogen.RESULTS: MSCs were isolated from human umbilical cord and presented with fibroblastic morphology. 80% of cells were at G_0/G_1 phase with good growth activity and stably passaged over 20 times. These cells were positive for CD29, CD105, and Vimentin, but negative for CD34 and CD31. MSCs were induced to hepatocyte-1 ike cells that were positive for alpha fetoprotein, CK18, CK19 at 1 week and albumin at 3 weeks. At 4 weeks, induced cells were positive for glycogen staining.CONCLUSION: MSCs isolated from human umbilical cord can be cultured in a long periods time in vitro and are able to differentiate into functional hepatocyte-like cells.

Objective To investigate the effect of human umbilical cord mesenchymal stem cell paracrine substance on proliferation and apoptosis of liver cells in vitro. Methods Mesenchymal stem cells (MSC)were separated from human umbilical cord with type Ⅳ collagenase and trypsogen digestion method and cultured in vitro. The human umbilical cord mesenchymal stem cells-conditioned medium(MSC-CM) which contain paracrine substance of human umbilical cord mesenchymal stem cells (HUCMSC) was prepared. Hepatocytes were isolated from SD rats by low concentration collagenase perfusion procedure. There were three groups in the experiment, control group, 2% MSC-CM group and 8% MSC-CM group. The proliferation of normal hepatocytes were assayed with MTT method. We detected the urea and albumin level in culture supernatant to assay the hepatocyte function under different concentration MSC-CM. Hepatocytes were induced for apoptosis by Actinomycin D and tumor necrosis factor alpha (TNF-α),and the apoptosis effect of different concentration MSC-CM was assayed with LIVE/DEAD Viability/Cytotoxicity Kit. Results The MTT assay showed that the absorbance of 2% MSC-CM group was significantly increased (P＜0. 01), and the urea and albumin levels of 2 % MSC-CM group were also significantly increased when compared with control group(P＜0. 01).LIVE/DEAD Viability/Cytotoxicity Kit revealed that hepatocyte survival rate of 2 % MSC-CM group was increased when compared with control group(P＜0. 05), there were no significant differences in above-mentioned experiments when 8% MSC-CM group compared with control group. Conclusion The low concentration MSC-CM could stimulate normal hepatocyte proliferation, inhibit impaired hepatocyte apoptosis and improve hepatocyte function.

Background and purpose: Triple negative breast cancer (TNBC) is a high risk breast cancer characterized by the negative expression of estrogen receptor(ER), progesterone receptor (PR) and Her-2 that have no specific therapy. This study was to analyze clinical pathological characteristics, survival, and prognostic factors of patients with TNBC. Methods: Clinical and pathological as well as follow-up data of TNBC, treated at the Cancer Centre of Sun Yat-sen University from Jan. 2000 to Dec. 2003, were collected and analyzed. Results: A total number of 128 women were identified as having triple negative breast cancer. The median age of these patients was 46 years, and 60.9% of them had stage Ⅰ or Ⅱ disease. The majority of pathological types were invasive ductal carcinomas, and 78.1% of tumors were staged T1 or T2. And 48.4% of these patients were involved in lymph node. Event-free survival, local replase-free survival, distant metastasis-free survival and overall survival at five years were 71.1%, 84.3%, 75.8% and 83.6% respectively. Though lymph node metastasis, tumor masses, stage and lymph-vascular invasion were all found to be related to overall survival, however, only lymph node metastasis and tumor masses affected the overall survival as revealed by the Cox proportional hazard model analysis. Conclusion: Triple negative breast cancer has distinct clinical and pathological characteristics. The patients are usually young, with large masses, lymph node metastasis, family history of breast cancer and poor prognosis; lymph node metastasis and tumor mass are important prognostic factors.

Objective To investigate the clinical application of serum interleukin-6 (IL-6)and interleukin-22 (IL-22) levels in predicting the recurrence of hepatitis B virus (HBV)-related early hepatocellular carcinoma (HCC) after receiving microwave ablation (MWA).Methods Preoperative peripheral blood samples were collected in 49 patients with early-stage HBV-related HCC,and serum concentrations of IL-6 and IL-22 were measured by using ELISA.Thirty healthy volunteers were recruited and used as the control group.The xtile software was used to define the best cut-off value,and the IL-6 and IL-22 levels were divided into highlevel group and low-level group.The tumor-free survivals of high-level and low-level groups were analyzed with Kaplan-Meier analysis,log rank test was adopted to determine the difference,and Cox regression model was employed to screen the risk factors affecting HBV-related HCC recurrence.Results The serum IL-6 and IL-22 levels of HCC group were 13.20 pg/ml (11.87-15.79 pg/ml) and 42.18 pg/ml (34.39-57.44 pg/ml) respectively,which were significantly higher than 10.47 pg/ml (9.50-13.82 pg/ml) and 25.45 pg/ml (22.31-30.12 pg/ml) of the control group (P=0.001 and P＜0.001 respectively).Kaplan-Meier analysis revealed that preoperative lower IL-6,higher total bilirubin and lower albumin levels indicated a shorter disease-free survival (DFS),and IL-22 levels had no statistically significant effect on the recurrence of HCC.Cox regression multivariate analysis showed that lower serum IL-6 level (≤ 13.2 pg/ml;hazard ratio=3.721;95％ CI=1.674-8.272;P=0.001) and lower serum albumin level (≤41.0 g/L;hazard mtio=2.085;95％CI=1.101-3.950;P=0.024) were independent risk factors affecting HBV-related HCC recurrence Conclusion Preoperative serum IL-6 level and serum albumin level can be used as the predictors of HCC recurrence in patients with HBV-related early HCC who are receiving MWA treatment.(J Intervent Radiol,2017,26:232-236)

Objective:To determine the diagnosis, treatment, and pathogenesis of ketamine-associated cystitis.
Methods:Clinical data from 3 patients with ketamine-associated cystitis were analyzed retrospectively and discussed in light of relevant literature.
Results:In the 3 cases, 2 presented severe lower urinary tract symptoms, including frequency, urgency, dysuria, urge incontinence, and painful haematuria. Urinalysis and urine culture were negative. Imaging examination demonstrated thickening of the bladder wall and a small capacity. Inflammatory changes in the bladder mucosa were observed by cystoscopy and biopsies. After cessation of ketamine use, with the addition of steroids or hydrodistension, the symptoms in the 3 patients improved. The symptoms recurred in 2 patients, as 1 was exposed to ketamine again and 1 had severe bladder contraction after for 3-4 month follow-up.
Conclusion:Ketamine-associated cystitis is a new urinary system inlfammatory damage. Its etiology and treatment methods are not clear. early abstinence from ketamine use and early treatment are crucial for patients with ketamine-associated cystitis to avoid irreversible damage.

Objective:To explore the effect of low intensity pulsed ultrasound (LIPUS) on TGF-β1/Smad 2,3 signal pathway during the dentin injury and repair.Methods:Among 25 Sprague-Dawley rats,5 rats served as a blank control group without treatment.The remaining 20 rats received modified caries preparation inbilateral maxillary first molars to establish a model of dentin-pulp injury and repair.The right maxillary first molars served as a LIPUS group,which received LIPUS irradiation (frequency:1.5 MHz,pulse width:200 μs,pulse repetition frequency:1 kHz,spatial averaged temporal averaged intensity:30 mW/cm2,20 min/d),and the left maxillary first molars served as a cavity-prepared group,which received fake LIPUS irradiation.The rats were sacrificed at 1,3,5,7 and 14 days after LIPUS irradiation.Immunohistochemical staining and Image-pro plus 6.0 were applied to detect the expression and distribution of transforming growth factor-beta1 (TGF-β1) and small mothers against decapentaplegic 2/3(Smad 2 and Smad 3).Results:Immunohistochemical staining showed that the expression of TGF-β1 and Smad 2,3were low innormal pulp,but they were increased in different degree after dentin injury.The result of image analysis showed that the expression of TGF-β1 in the cavity-prepared group gradually increased at the first day and peaked at day 5,and then it returned to normal level at day 14.However,the expression of TGF-β 1 in the LIPUS group were significantly higher than that in the cavity-prepared group at day 3 and 5 (bothP＜0.05).The expressions of Smad 2,3 in both the LIPUS group and the cavity-prepared group were consistently increased all the way, but the expressions in the LIPUS group were higher compared with that in the cavity-prepared group (P＜0.05).Conclusion:The TGF-β1/Smad 2,3 signal pathway can be activated during the dentin injury and repair.LIPUS can up-regulate the expression of TGF-β1 and Smad 2,3 in the early period,which may take part in the dentin-pulp complex injury and repair process.