·AIM: To study the effect of an intravitreal injection of angiostatin on vascular leakage in the retina and iris of oxygen-induced retinopathy of prematurity (ROP).·METHODS: Brown Norway rats at postnatal day 7 (P7) were exposed to hyperoxia (750mL/L O2 )for 5 days (P7-12) and then returned to normoxia to induce retinopathy. Angiostatin was reconstituted in sterile Phosphate Buffered Saline(PBS) and diluted to desired different concentrations. Angiostatin solution was injected into the vitreous of the right eye of the ROP rats at P14 and the age-matched normal rats through pars plana using a glass capillary, and the left eye received the same volume of sterile PBS as the control. Vascular permeability was quantified at 1, 2 and 3 days after the injection by measuring albumin leakage from blood vessels into the retina and iris using the Evans blue method and normalized by total protein concentrations. The expression of vascular endothelial growth factor (VEGF) in retina was evaluated using the Western Blot analysis and immuno-histochemistry 24 hours following the injection.·RESULTS: ROP rats showed significant increases of vascular permeability in the retina and iris (P<0.01). Angio-statin reduces vascular permeability in a dose-dependent manner in the retina of ROP rats. The reduction showed a time course trend. [Angiostatin injection reduced retinal vascular permeability by approximately 1.5 and 2-fold at P15 (P<0.05) and P16 (P<0.01), respectively.] Angiostatin injection significantly reduced VEGF levels in the retina of ROP rats but did not affect retinal VEGF levels in normal rats.·CONCLUSION: Angiostatin significantly decreases pa-thological vascular permeability in the retina and iris of ROP rats but not in normal rats. Angiostatin down-regulates VEGF expression in retina of ROP rats. These results suggest that angiostatin may have a therapeutic potential in the treatment of ROP and other diseases with vascular leakage.

The aim of this study was to fabricate biomatrix/polymer hybrid scaffolds using an electrospinning technique. Then tissue engineered heart valves were engineered by seeding mesenchymal stromal cells (MSCs) onto the scaffolds. The effects of the hybrid scaffolds on the proliferation of seed cells, formation of extracellular matrix and mechanical properties of tissue engineered heart valves were investigated. MSCs were obtained from rats. Porcine aortic heart valves were decellularized, coated with poly(3-hydroxybutyrate-co-4-hydroxybutyrate) using an electrospinning technique, and reseeded and cultured over a time period of 14 days. In control group, the decellularized valve scaffolds were reseeded and cultured over an equivalent time period. Specimens of each group were examined histologically (hematoxylin-eosin [HE] staining, immunohistostaining, and scanning electron microscopy), biochemically (DNA and 4-hydroxyproline) and mechanically. The results showed that recellularization was comparable to the specimens of hybrid scaffolds and controls. The specimens of hybrid scaffolds and controls revealed comparable amounts of cell mass and 4-hydroxyproline (P>0.05). However, the specimens of hybrid scaffolds showed a significant increase in mechanical strength, compared to the controls (P<0.05). This study demonstrated the superiority of the hybrid scaffolds to increase the mechanical strength of tissue engineered heart valves. And compared to the decellularized valve scaffolds, the hybrid scaffolds showed similar effects on the proliferation of MSCs and formation of extracellular matrix. It was believed that the hybrid scaffolds could be used for the construction of tissue engineered heart valves.

Objective To review the existing literature and quantitatively evaluate the association of circulating vaspin levels and the risk of gestational diabetes mellitus ( GDM) ． Methods We systematically searched the PubMed，EMBASE，Web of Science，China National Knowledge Infrastructure，and WanfangData databases up to June 2019． Pooled standardized mean differences ( SMDs) with 95% confidence intervals ( CIs) were calculated using random－ or fixed－effects models based on the heterogeneity of studies． Subgroup analyses，Meta－regression，sensitivity and publication bias were assessed to analyze the heterogeneity and the robustness of the results． All statistical analyses were performed using STATA 12．0． Ｒesults Nine articles ( 11 comparisons) published from 2013 to 2019 were included in our final Meta－analysis，covering a total of 738 patients with GDM and 661 normal pregnant women． There was significant difference in the overall maternal circulating vaspin levels between GDM patients and healthy pregnant women ( SMD= 0．613，95% CI: 0．044－1．182，P= 0．035) ． Subgroup analyses stratified by trimester in which vaspin was measured and whether BMI was matched suggested the similar trend to the overall result． Subgroup analysis according to ethnicity found that circulating vaspin level might not be related to GDM in " European" subgroup; sensitivity analysis by excluding moderate－quality studies and BMI－unmatched studies found that circulating vaspin levels were still related to GDM risk． Conclusions Our Meta－analysis indicated that maternal circulating vaspin levels might be positively correlated with the risk of GDM in Asians．

OBJECTIVETo assess the anti-tumor immune response to percutaneous cryoablation in patients with local prostate cancer.

METHODSWe treated 10 patients with local prostate cancer by percutaneous cryoablation, collected the blood samples before and 2 weeks after the treatment and isolated peripheral blood mononuclear cells (PBMCs). Protein lysates were made by biopsy from autologous prostate cancer or non-cancer tissues. The levels of serum TNF-alpha, IFN-gamma, IL4 and IL-10 were determined by enzyme-linked immunosorbent assay (ELISA) and the Th1/Th2 ratio was calculated by the IFN-gamma/IL-4 ratio. The number of IFN-gamma + T cells under the stimulation of different protein lysates was counted by enzyme link immunol spot (ELISPOT). And the cytolytic activity of cytotoxic T lymphocytes (CTL) was detected by LDH assay.

RESULTSCompared with pre-treatment, the levels of TNF-alpha and IFN-gamma, the Th1/ Th2 ratio and the number of IFN-gamma + T cells induced by tumor protein lysates in PBMCs were increased significantly after cryosurgery (P < 0.01), while the levels of IL4 and IL-10 decreased slightly, and the non-tumor protein lysates induced no obvious changes in the number of IFN-gamma T cells. The cytolytic activity of cytotoxic T lymphocytes against human prostate cancer cells LNCaP was markedly increased, but not that against renal cancer cells GRC-1. One case of recurrence was found during the 3-6 months follow-up.

CONCLUSIONPercutaneous cryoablation for prostate cancer could induce a tumor-specific immune response.

Aged ; Cryosurgery ; Humans ; Interferon-gamma ; blood ; Interleukin-10 ; blood ; Interleukin-4 ; blood ; Male ; Middle Aged ; Prostatic Neoplasms ; immunology ; therapy ; T-Lymphocytes, Cytotoxic ; immunology ; Th1 Cells ; immunology ; Th2 Cells ; immunology ; Tumor Necrosis Factor-alpha ; blood

This study was to investigate dynamics of biological properties of CD133(+) cells from human umbilical cord blood (UCB) during short-term culture containing the combination of hematopoietic growth factors and the feasibility of in vitro expansion of CD133(+) cells. The biology activities including analysis of cell cycle, immunophenotype, telomerase activity, expression of adhesion molecules and expansion potential of CD133(+) cells were monitored during ex-vivo expansion, and compared with those of CD34(+) cells. The results showed that the contents of CD133(+) and CD34(+) cells in fresh UCB were (1.05 +/- 0.73)% and (1.40 +/- 0.56)% respectively. About 79.62% of CD34(+) cells expressed CD133, and more than 97% of CD133(+) cells were CD133(+)CD34(+), markedly higher than that in CD34(+) fraction (P < 0.01). No significant differences were observed in content of cells expressing CD38, CD13, CD14, CD61 and glycophorin-A between the two fractions. Expansion of CD133(+), CD133(+)CD34(+) and CD34(+)CD38(-) cells at 10 days and those of CFU-mix, HPP-CFC and CD34(+)CD38(-) cells at 6 days from CD133(+) cells group were significantly higher than those from the CD34(+) cell group (P < 0.05). Analysis of immunophenotype showed that CD133(+)CD34(+) cells declined gradually while CD133(-)CD34(+) and CD133(-)CD34(-) cells increased during ex-vivo expansion; basal telomerase activities of fresh UCB CD133(+) and CD34(+) cells were low but significantly exceeded that of CD34(-) fraction (P < 0.05). At first week of expansion, telomerase activity was significantly upregulated, after two weeks, telomerase activity remarkably declined, and decreased to baseline or below the limits of detection in day 20. More than 90% of CD133(+) cells expressed CD49d and CD11a, and, more than 85% of the cells expressed CD54, about 50% of cells expressed CD62L. At the early stage of expansion, expression of CD49d was upregulated, expression of CD11a remaining no change, while as expression of CD54 and CD62L was downregulated. Expression of all adhesion molecules was decreased gradually with extend of culture. But expression of these adhesion molecules on CD34(+) subsets were not affected significantly during expansion. It is concluded that CD133(+) population may be a more primitive hematopoietic stem/progenitor cells (HSPC) than CD34(+) cells, CD133(+) cells have great expansion potential for ex-vivo expansion and is a suitable target cell for ex-vivo expansion of HSPC. Downregulation of adhesion molecules and telomerase activity may be one of the reasons for delayed engraftment of expanded products.
AC133 Antigen ; Antigens, CD ; Antigens, CD34 ; analysis ; Cell Cycle ; Cells, Cultured ; Fetal Blood ; cytology ; Glycoproteins ; analysis ; Hematopoietic Stem Cells ; physiology ; Humans ; Immunophenotyping ; Peptides ; analysis ; Telomerase ; metabolism

Objective To evaluate the preventive and therapeutic effect of oral solution of Niao Du Kang (尿毒康) on acute renal tubular necrosis (ATN) in Sprague-Dawley (SD) rats,and preliminarily approach its mechanisms.Methods The oral solution was composed of traditional Chinese medicinal herbs of Radix et Rhizoma Rhei (大黄),Radix Sanguisorbae Offieinalis (地榆),Radix Salviae Miltiorrhizae (丹参), Radix Astragali seu Hedysari (黄芪),Flos Carthami Tinctorii (红花) and so on.Sixty-four SD male rats were randomly divided into 4 groups:normal control group,ATN model group,verapamil treatment group and traditional Chinese medicine,oral solution of Niao Du Kang treatment group,every group having 16 rats.. The ATN model of SD rats was induced by intramuscular injection of 50% glycerin mixed with isotonic saline solution at the back of bilateral lower extremities.After the model was established in each group ,the levels of blood urea nitrogen (BUN),levels of blood serum creatinine (SCr),renal failure index (RFI) and renal pathological changes were detected at different time points after modeling for 12 and 24 hours,and the therapeutic effect of oral solution of Niao Du Kang was observed.Results After the model establishment, Niao Du Kang oral solution could lower the elevation of blood BUN,SCr,RFI in the model,and compared with the normal control group,there were statistical significances (all P

OBJECTIVETo describe observation of security and availability of covering left subclavian artery during thoracic endovascular aortic repair (TEVAR) in follow-up.

METHODSFrom December 2007 to December 2008, 111 consecutive patients received stent grafts to treat lesions involving thoracic aorta. According to the covering of left subclavian artery, four groups including total covering (TC), less-than 50% covering (LTC), more-than 50% covering (MTC) and non-covering (NC) were formed. Difference of blood pressure between two upper extremities was required before TEVAR and 1st, 3rd, 5th, 30th day after TEVAR. Patients were evaluated postoperatively and at follow-up for stroke as well as symptoms of paraplegia, hemiparalysis or left upper extremity claudication.

RESULTSFifty-five (49.6%), 18 (16.2%), 7 (6.3%) and 31 (27.9%) cases were divided into TC, LTC, MTC and NC groups, respectively. Difference of blood pressure between TC and the 3 latter groups were significantly different (P<0.01). Complications appeared as followed during one week after TEVAR: 13 patients in dizziness, among which 5 patients suffered from amaurosis and spotted vision, and 7 patients in left upper extremity claudication. No stroke, paraplegia or hemiparalysis in TC. Thoracic aortic lesions were successfully excluded in all patients.

CONCLUSIONSIntentional coverage of left subclavian artery to obtain an adequate proximal landing zone during TEVAR is safe and well-tolerated. But it may be managed expectantly with some exceptions for further lasting efficacy.

Aorta, Thoracic ; surgery ; Aortic Aneurysm, Thoracic ; surgery ; Blood Vessel Prosthesis Implantation ; Female ; Humans ; Male ; Middle Aged ; Prospective Studies ; Stents ; Subclavian Artery ; surgery ; Treatment Outcome

OBJECTIVETo study apoptosis and gene FasL expression induced by carbon disulfide in sertoli cells of male rats.

METHODSSertoli cells were exposed to different concentrations of CS(2) (0, 0.36, 0.72, 1.44 micromol/ml) for 24 hours. Survival rate, apoptosis rate, expression level of gene FasL were measured using MTT, FCM, and RT-PCR methods respectively.

RESULTSSertoli cell survival rate decreased as the concentration of CS(2) increased. The survival rate (73.34% +/- 1.39%) was significantly lower than the control group (99.98% +/- 5.48%) when the concentration of CS(2) > or = 1.44 micromol/ml (P < 0.05). Apoptosis rate increased as the CS(2) concentration increased. Apoptosis rate (7.93% +/- 0.43%) was significantly higher when the concentration of CS(2) > or = 1.44 micromol/ml (P < 0.05). Expression level of the FasL significantly increased as the concentrations of CS(2) (P < 0.05).

CONCLUSIONCS(2) is cytotoxic to sertoli cells. It could cause apoptosis of sertoli cells.

Animals ; Apoptosis ; drug effects ; Carbon Disulfide ; toxicity ; Cell Line ; Cell Survival ; Fas Ligand Protein ; metabolism ; Male ; Rats ; Sertoli Cells ; drug effects ; metabolism ; Testis ; cytology

OBJECTIVETo observe the expressions of nestin and glial fibrillary acidic protein (GFAP) and their association with reactive astrocytes following spinal cord injury in adult rats.

METHODSAdult rats with compression injury of the spinal cord were divided into 7 groups (n=6) and examined at 1, 3, and 5 days and at 1, 2, 4 and 8 weeks after the injury. The recovery of the locomotor function after the injury was evaluated with Basso, Beattie and Bresnahan (BBB) scale, and the degree and scope of the spinal injury were assessed using toluidine blue staining. Immunohistochemistry, double immunofluorescent labeling and an image analysis system were employed to observe nestin and GFAP expression and cell proliferation in different regions of the spinal cord.

RESULTSThe bilateral hind limb locomotor function of the rats declined severely 24 h after the spinal cord injury and underwent substantial recovery in 1 or 2 weeks after the injury, but followed by rather slow recovery afterwards. Toluidine blue staining of the spinal cord 24 h after the injury showed significant pathological changes in the neurons. The extension of the tissue injury increased with time till 1 week after the spinal cord injury. The site of injury and the adjacent tissues presented with markedly increased nestin and GFAP expressions 24 h after the injury, and nestin+/GFAP(-) cells dominated in the ependymal region around the central canal, whereas nestin+/GFAP+ dominated in the in other regions, showing significant difference from the control group. Nestin and GFAP expression reached the peak level 3 to 7 days after the injury and declined gradually till reaching nearly the control level at 2 weeks.

CONCLUSIONCompression injury of the spinal cord induces up-regulated expressions of nestin and GFAP, and nestin expression is positively correlated to the reactive astrocytes, which, along with the neural stem cells, respond to spinal nerve injury and possibly play a role in repair of the central nervous system injury.

Animals ; Astrocytes ; pathology ; Glial Fibrillary Acidic Protein ; biosynthesis ; genetics ; Intermediate Filament Proteins ; biosynthesis ; genetics ; Male ; Nerve Tissue Proteins ; biosynthesis ; genetics ; Nestin ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Spinal Cord Injuries ; metabolism ; pathology ; Stem Cells ; cytology ; metabolism ; Up-Regulation

Objective Circulating tumor cells (CTCs) have potential value in the clinical application of various tumors. This study was to investigate the role of CTCs and their chemokine receptor CCR9 in the invasion and metastasis of non-small cell lung cancer (NSCLC). Methods From May 2018 to June 2019, a total of 62 patients with NSCLC in the clinical oncology center of The People's Hospital of Guangxi Zhuang Autonomous Region were enrolled in this study. The CanpatrolTM CTC technique was used to detected the expressions of CTCs and CCR9 in CTCs in peripheral blood of patients. Furthermore, the relationships between expression levels of CTCs, CCR9 and clinical, pathological characteristics of NSCLC patients were analyzed. Results CTCs were detected in 56 of 62 (90.3%) NSCLC patients. CTCs counts were associated with TNM stage, lymph node metastasis and distant metastasis of NSCLC (P<0.05). In the analysis of clinical correlation between CTC subtypes and NSCLC, epithelial CTCs counts were related to TNM stage and distant metastasis of NSCLC (r=0.296 and r=0.273, P<0.05). Additionally, counts of mixed type CTCs were also correlative with NSCLC tumor metastasis (r =0.253, P =0.047). Finally, we found that the positive rate of CCR9 in mixed type CTCs was associated with distant metastasis of NSCLC (r=0.353, P=0.038). Conclusion CTCs counts and subtypes were correlated with TNM stage and metastasis of NSCLC. The expression level of CCR9 on CTC was expected to be a biomarker to evaluate the risk of tumor metastasis in NSCLC.