Objective To determine the effects of angiotensin Ⅱ (Ang Ⅱ) on NF-κB DNA binding activity in alveolar macrophage. Method Human alveolar macrophages were isolated and made homogeneous from alveo-lar lavage fluid, and cuhtured in DMEM. Alvcolar macrophages were treated with AugⅡ (10-6M) for 15 min, 30 min, 60 min and 120 min, respectively. Moreover, alveolar macmphages were pretreated with irbesartan (AngⅡ type 1 receptor blocker) for Ⅰ hour before stimulated with Angiotensin Ⅱ for Ⅰ hour. Electrophoretic gel mobility shift assay (EMSA) was used to detect NF-κB DNA binding activity. The protein expression of IκBα was examined by Western blot. Expressions of TNF-α and ICAM-1 mRNA were detected by using RT-PCR. Results EMSA re-vealed that there was a increase in up-regulation of NF-κB DNA binding activity after alveolar macrophages were treated with Ang Ⅱ for 15 rain and peaked at 60 min. Irbesartan treatment reduced DNA binding activity. Com-pared with control group, the protein expression of IκBα decreased in Ang Ⅱ treatment group(0.29±0.11, P= 0.013), and Irbesartan treatment significantly increased protein expression of IκBα(0.83±0.12, P=0.001). The expressions of TNF-α and ICAM-1 mRNA were up-regulated by AngⅡ in comparison with the control group (TNF-α:1.13±0.17 vs. 0.42±0.099; ICAM-1 0.55±0.08 vs. 0.16±0.050, P=0.003). Irbesartan inhibited the expressions of TNF-α (0.77±0.15 vs 1.13±0.17, P=0.02; ICAM-1(0.32±0.07 vs 0.55±0.08, P =0.001). Conclusions Ang Ⅱ is capable to stimulate NF-κB signal pathway in alveolar macrophages.

AIM: To investigate the preparation and parameter of pellets of traditional Chinese medicine in order to solve the problem of preparation of pellets of traditional Chinese medicine. METHODS: We took centrifugal spray method to prepare the pellets, and compared effect of spray coating and fluidzed bed coating on the quality of pellets. RESULTS: The best preparation of pellets was determined as followed. The frequency of turntable was 45 HZ, the flow rate of liquid was 1.2 g/min. the relative density of liquid was 1.20 g/cm 3; when coating weigh reached 14%, the better pellets could be obtained. CONCLUSION: The need of the assistant matter of taking spray pellets and film coating was less and roboticized. It accorded with the answer of GMP and the regular production of the preparation of pellets of traditional Chinese medicine.

Endothelial progenitor cells (EPCs) are immature endothelial cells which have the capacity to proliferate, migrate and differentiate into mature endothelial cells from bone marrow to the peripheral circulation. EPCs have been shown to participate in postnatal endothelial repair and neovascularization of ischemic organs, and have been used as a new source of seeded cells in vascular tissue engineering. In this review, we focus on the origin, identification, property and function of EPCs as well as their application in vascular tissue engineering.
Animals ; Blood Vessels ; physiopathology ; Endothelial Cells ; cytology ; physiology ; Endothelium, Vascular ; pathology ; physiology ; Humans ; Neovascularization, Physiologic ; physiology ; Recovery of Function ; physiology ; Stem Cells ; cytology ; physiology ; Tissue Engineering ; methods

OBJECTIVETo summarize the features of disease history, clinical manifestations, adjuvant examination results, diagnosis, treatments and outcome of patients with histoplasmosis.

METHODSThe clinical data of 14 patients with biopsy-confirmed histoplasmosis between 2000 and 2012 in Nanfang Hospital were analyzed retrospectively.

RESULTSThe clinical manifestations of histoplasmosis included fever, productive cough, chest pain, and abdominal pain, accompanied occasionally by neurological symptoms, lymph node enlargement or surface mass. Seven out of the 14 o patients had underlying immunosuppressive conditions, 9 had chest imaging changes, and 2 showed reduced white blood cells, red blood cells and platelets. The cases were initially diagnosis as tuberculosis, malignant tumor, or malignant lymphoma before the definite diagnosis was established pathologically. Ten patients received treatments with itraconazole, amphotericin B, fluconazole or voriconazole, and 9 of them responded favorably to the treatments.

CONCLUSIONHistoplasmosis, with a low incidence and diverse clinical manifestations, presents with no specific imaging features to easily cause misdiagnosis and missed diagnosis, and its definite diagnosis relies on pathological examination.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Histoplasmosis ; diagnosis ; pathology ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

BACKGROUND:It is still a research focus on constructing substitution of the human tissues and organs, or producing the alliance for grafting by engineering methods in tissue engineering. Among these researches, it is pivotal to choose appropriate materials. The prepared scaffolds should have suitable tensile strength and mechanical toughness to withstand the various outside forces without being damaged. So, it is very necessary to evaluate the biomechanical properties of candidated materials in tissue engineering, which can supply the references for selecting materials for tissue scaffolds and their designation.OBJECTIVE: To investigate the biomechanical properties of nine kinds of scaffold materials, in order to supply a biomechanical basis for the selection and design of scaffold materials for tissue engineering.DESIGN: A repetitive measurement study.SETTING: College of Bioengineering, Chongqing University.MATERIALS: The materials involved in this study were poly (DL-lactic-co - glycolic acid) (PLGA), sodium polymannuronate, gelatine, chitosan, collagen, acellular porcine dermis (APD), acellular vascular matrix (AVM),APD-PLGA, AVM-PLGA, modified gelatine and chitosan.METHODS: All the experiments related to this study were completed in the Biorheology laboratory of the College of Bioengineering, Chongqing University from April 2006 to March 2007. The nine materials above were prepared, gelatine and chitosan were modified. Stress-strain testing was performed at 10 mm per minute by a material testing machine (INSTRON 1011, USA). The Yang's modulus was calculated in the range of 0.005 to 0.02, the ultimate strain and stress were also obtained.MAIN OUTCOME MEASURES: The ultimate strain, ultimate stress and Yang's modulus of the nine materials were analyzed.polymannuronate ＞ AVM-PLGA ＞ collagen ＞ gelatine (P ＜ 0.05). The rate of burst straining of chitosan and PLGA were greater than those of other materials, 133% and 276% respectively (P ＜ 0.05). In addition, after being combined with ultimate stresses of APD and APD-PLGA were greater than that of other materials, i.e., their burst strengths were greater than those of other materials. The data also indicated that the burst strength of APD-PLGA was a little greater than that of APD (P ＞ 0.05). The burst strengths of gelatin, chitosan, and collagen were similar at the range of 7.67 to 9.51 MPa (P ＞ 0.05). The burst strengths of collagen and sodium polymannuronate were from 1.16 to 1.40 MPa, which were the least among all the materials. At the same time, being combined with PLGA, the burst strength of AVM-PLGA greatest, i.e., its rigidity was the greatest. The rigidity of APD was the least. After combined with PLGA, the rigidity of AVM and APD were increased (P ＜ 0.05), and corresponded with PLGA (P＞ 0.05). Except for gelatin, the order of rigidity in the materials was AVM-PLGA ＞ PLGA ＞ APD-PLGA ＞ AVM ＞ chitosan ＞ sodium polymannuronate ＞ collagen ＞ APD.CONCLUSION: AVM and APD have good biomechanical properties, which are very different from the water-soluble collagen. It is promising to improve the biomechanical properties of sodium polymannuronate, gelatin and chitosan by the complex of PLGA.

In this study we examined the in vitro characteristics of tenocyte adhesion to biologically-modified surface of polymer. Polylactic-co-glycolic acid (PLGA) 85/15 films were prepared by a solvent-casting technique. Each film was adhered onto the bottom of a chamber. The film was precoated with poly-D-lysine (PDL), and then coated with serum-free F12 medium containing various concentrations of fibronectin (FN), type I collagen (CN I), and insulin-like growth factor1 (IGF-1). The monoclonal antibodies (to FN and to CN I) with various dilutions were used to inhibit attachment of tenocytes to surface precoated with FN or CN I. Human embryonic tendon cells (HETCs) and transformed human embryonic tendon cells (THETCs) were used as the seeding cells. The system used for the measurement of adhesion force was the micropipette aspiration experiment system. The micropipette was manipulated to aspirate a small portion of the tenocyte body by using a small aspiration pressure. Then the pipette was pulled away from the adhesion area by micromanipulation. The minimum force required to detach the tenocyte from the substrate was defined as the adhesion force. The results showed that modification of FN or CN I by precoating significantly enhanced attachment of tenocytes to surface of polymer (P < 0.05). As antibodies to FN or CN I were added to a polymer film precoated with FN or CN I, the adhesion force decreased significantly (P < 0.05). We concluded that the specific adhesion forces of tenocytes to extracellular matrix adhesion proteins (FN and CN I) had coordinated action and showed good dependence on their precoating concentrations, and were inhibited by the antibodies to these adhesion proteins. Films precoated with IGF-1 strongly accelerated the adhesion of tenocytes to polymer. These results indicate that the specific adhesion of tenocytes to polymer can be promoted by coating extracellular matrix adhesive proteins and insulin-like growth factor1. It is of great importance to construct tissue-engineered tendon.
Biocompatible Materials ; chemistry ; Cell Adhesion ; drug effects ; physiology ; Cells, Cultured ; Extracellular Matrix Proteins ; pharmacology ; Growth Substances ; pharmacology ; Humans ; Lactic Acid ; chemistry ; Polyglycolic Acid ; chemistry ; Polylysine ; pharmacology ; Polymers ; chemistry ; Tendons ; cytology ; embryology ; physiology ; Tissue Engineering

A few of flow experiments were utilized to verify a theoretical hypothesis proposed by Fung and coworker which showed that the tensile stress in the upper cell membrane of the vascular endothelium could accumulate upstream against the direction of blood flow. Endothelial cells from replicate human umbilical vein segment (HUVSEC) in vitro with length of 11 cm and 21 cm were exposed to the same pulsatile laminar shear stress averaged of 0.12 N/m2 for 42 hours. The average production rate of endothelin-1(ET-1), at 11 cm segment is 50% lower than that at 21 cm segment(16.93 +/- 0.89) vs. (26.13 +/- 1.79) pg/cm2.h respectively. The average production rate of ET-1 under pulsatile laminar flow was significantly higher than that under steady laminar flow. It showed that, high correlation of the length of HUVSEC with their ET-1 metabolism exists, suggesting that the tensile stress in the upper endothelial cell membrane could accumulate.
Cell Membrane ; physiology ; Endothelin-1 ; metabolism ; Endothelium, Vascular ; cytology ; physiology ; Humans ; In Vitro Techniques ; Infant, Newborn ; Stress, Mechanical ; Umbilical Veins ; anatomy & histology

OBJECTIVETo construct a lentiviral expression vector for human VHL and its shRNA vector, and study the effect of VHL on proliferation and apoptosis of renal cell carcinoma cell lines.

METHODSLentiviral vectors pZsGreen1-VHL and pLL3.7-shVHL were constructed and transfected into 293T cells with 3 packaging plasmids by Lipofectamine(TM) 2000 reagent. The supernatant was collected to infect A498 and Caki-1 cells, respectively. VHL mRNA and protein levels were detected by RT-PCR and Western blotting, respectively. The effect of VHL on the proliferation, cell cycle and cell apoptosis were analyzed by MTS and flow cytometry.

RESULTSThe recombinant lentiviral vectors were successfully constructed. The proliferation of A498 cells with reconstituted wild-type VHL was significantly inhibited, while the proliferation of Caki-1 cells with VHL knockdown was significantly enhanced as compared with the control cells (P<0.05). VHL induced G0/G1-S cell cycle arrest. The apoptosis rate of A498 cells with reconstituted wild-type VHL was significantly increased while that of Caki-1 cells with VHL knockdown was significantly lowered compared with the control cells (P<0.05).

CONCLUSIONVHL can inhibit the proliferation and induce apoptosis of renal cell carcinoma cells.

Apoptosis ; Carcinoma, Renal Cell ; pathology ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Lentivirus ; Plasmids ; RNA, Messenger ; RNA, Small Interfering ; Transfection ; Von Hippel-Lindau Tumor Suppressor Protein ; genetics

OBJECTIVETo investigate the expression of miR-223 in clear cell renal cell carcinoma (ccRcc) and its clinical implications.

METHODSQuantitative real-time PCR was employed to detect the levels of miR- 223 expression in ccRcc, pair-matched adjacent normal tissues and different renal cancer cell lines. Transwell migration essay and wound healing essay were used to evaluate the invasion and migration of renal cancer 786-O cells transfected with miR-223 mimics. MTT essay was used to measure the cell proliferation, and the cell cycle changes following the transfection were analyzed with flow cytometry.

RESULTSCompared with the normal tissues, the cancer samples showed up-regulated miR-223 expression, which was associated with tumor size. In 786-O cell cultures, transfection with miR-223 mimics significantly enhanced cell migration (P<0.0001) and growth (P=0.006) and induced G1 cell cycle arrest.

CONCLUSIONmiR-223 promotes renal cancer cell migration and proliferation and may serve as a potential therapeutic target for ccRcc.

Carcinoma, Renal Cell ; metabolism ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Flow Cytometry ; G1 Phase Cell Cycle Checkpoints ; Humans ; MicroRNAs ; metabolism ; Real-Time Polymerase Chain Reaction ; Transfection ; Up-Regulation

Objective:To investigate the characteristics of 18F-DCFPyL PET/CT imaging in castration-resistant prostate cancer (CRPC) patients with different PSA levels. Methods:The imaging and clinical data of 50 patients with CRPC who underwent 18F-DCFPyL PET/CT examination in Chinese PLA General Hospital from January 2018 to December 2020 were analyzed retrospectively. The average age was 72 (54-95) years old. Serum total PSA was 92.28(0.36-2000.00) ng/ml. According to the total PSA level, the patients were divided into low PSA group(total PSA ≤ 1 ng/ml, n=9), medium PSA group (1 ng/ml10ng/ml, n=23). According to the standardized evaluation standard of molecular imaging, the suspicious tumor lesions on 18F-DCFPyL PET/CT imaging were scored by molecular imaging PSMA(miPSMA), and the miPSMA score ≥2 was defined as positive lesions. According to the number of lesions displayed by 18F-DCFPyL PET/CT, patients were divided into oligofocal group (the number of lesions ≤3) and multiple lesions group (the number of lesions >3). The imaging characteristics of patients in different groups were summarized. Results:The 18F-DCFPyL PET/CT imaging results of 50 cases in this study were all positive, including oligofocal group (n=27) and multiple lesions group (n=23). Of the 30 patients with unresected prostate, 18 had local recurrence of the prostate, while the other 12 patients with unresected prostate and 20 patients with resected prostate had no signs of local recurrence. The oligofocal group showed local recurrence, regional lymph node metastasis or bone metastasis. Patients with multiple lesions showed multiple lymph nodes and/or bone metastasis with or without local recurrence. There were 9, 18 and 4 patients with oligofoci in low, middle and high PSA groups, respectively.There were 27 patients in the oligonucleogenous group, and 21 of the 22 patients receiving local treatment were effective. All 3 patients treated with systemic treatments were effective. PSA progressed in 2 untreated patients. In the multi-foci group of 23 patients, 6 of 9 patients treated with abiraterone were effective. Two patients treated with enzalumide were ineffective. One of the 4 patients with chemotherapy was effective. One of the two patients treated with 177 Lu-PSMA nuclide was effective. One case did not respond to treatment with 89SrCl 2. Radiotherapy failed in 2 cases. PSA progressed in 3 untreated patients. Conclusions:18F-DCFPyLPET/CT imaging has a high detection rate of lesions in patients with CRPC and has potential guiding significance for follow-up treatment. The number of lesions in CRPC patients with different PSA levels was different, and the patients with low PSA levels were mainly oligofoci.