AIM:To investigate the effects and mechanism of anti-vascular endothelial growth factor ( VEGF) assisted pars plana vitrectomy ( PPV) in the treatment of proliferative diabetic retinopathy ( PDR) . ·METHODS: A total of 92 patients ( 92 eyes ) with PDR treated by PPV were divided into the simple PPV group (41 patients with 41 affected eyes) and the combined treatment group ( 51 patients with 51 affected eyes ) according to whether the patient underwent intravitreal injection of Ranibizumab ( IVR) . The combined treatment group was treated with IVR at 5-7d before PPV. The surgical time, times of electrocoagulation, silicone oil filling rate, the incidence of postoperative complications, LogMAR BCVA of affected eyes, levels of VEGF and pigment epithelium derived factor ( PEDF ) in aqueous humor and vitreous body were compared between the two groups. ·RESULTS:The surgical time was shorter, the times of electrocoagulation was less, the silicone oil filling rate and the incidence rates of iatrogenic retinal hole and vitreous body hematocele were lower in the combined treatment group than in the simple PPV group (P<0. 05). Levels of VEGF and PEDF in aqueous humor and vitreous body of the combined treatment group during PPV were lower than those in the simple PPV group (P<0. 05). The LogMAR BCVA of the affected eyes of the combined treatment group in 3mo after surgery was better than that of the simple PPV group (P<0. 05). ·CONCLUSION:IVR combined with PPV can reduce the perioperative levels of VEGF and PEDF, reduce the times of electrocoagulation and the incidence of iatrogenic retinal hole and vitreous body hematocele, and improve the visual acuity of patients with PDR.

CD40 and its receptor CD40L are a very important pair of co-stimulating molecule in immune response, which have extensive biological effects. After stimulating CD40 signal, it can exert corresponding function through MAPK (JNK, ERK, p38) pathway, PI3K cascade, as well as NF-κB and STAT. The CD40 signal is closely related to tumor immunity, this moleculer has already become targeted-molecule for cancer treatment. Recently, there have been many anti-CD40 monoclonal antibodies displaying good anti-cancer effect, among which CHIR-12.12, SGN-40 and CP-870, 893 developed rapidly and successively have entered clinical research stage. This review focuses the status of anti-CD40 monoclonal antibody, including distribution of CD40, physiological function of CD40, CD40 and tumor immunity, anti-CD40 monoclonal antibodies and so on.
Animals ; Antibodies, Monoclonal ; CD40 Antigens ; immunology ; Humans ; Neoplasms

Objective:To investigate the underlying mechanism of histone deacetylase (HDAC) inhibitor FK228-in- duced apoptosis of the prostate cancer cell line DU145.Methods:The inhibitory effect of FK228 on DU145 cell growth and its cytotoxicity were determined by MTT assay;cell cycle arrest was detected by flow cytometry assay;morphological change was observed by Giemsa staining;and defined kinase protein levels were determined by Western blot analysis.Re- suits:FK228 obviously inhibited DU145 cells growth,arrested cell cycle at G_0/G_1 phase,induced cells morphological changes and degraded several kinase proteins,including EGFR,Her2,Raf-1,Src,Cdk4 and IAP member Survivin.The degradation of these kinases blocked Raf-Mek-Erk and PI3K/Akt survival signal pathways,inducing apoptosis.Condu- sion:FK228 may induce DU145 cell apoptosis through depletion of multiple kinase proteins and blockade of survival sig- nal pathways of DU145 cells.

To establish methods and requirements for quality control of rhLFA3-IgG1, biological potency of rhLFA3-IgG1 was determined by CD2 molecule competitive binding assay on Jurkat cell surface. Purity of rhLFA3-IgG1 was analyzed by SEC-HPLC and IEC-HPLC. Peptide mapping was preformed by tryptic digestion and RP-HPLC after sample reduced and carboxymethylation by DTT and indoacetic acid, respectively. CHO host cell protein and Protein A residual were detected by ELISA separately. The quality control methods and requirements, such as biological potency, the physical-chemical characteristic of rhLFA3-IgG1 had been established. The methods and requirements for quality control of rhLFA3-IgG1 showed advantages of assuring the products safety and efficacy, which can be used for routine quality control of rhLFA3-IgG1.
Binding, Competitive ; Biotechnology ; methods ; CD2 Antigens ; metabolism ; CD58 Antigens ; biosynthesis ; chemistry ; Chromatography, High Pressure Liquid ; Humans ; Immunoglobulin G ; biosynthesis ; chemistry ; Jurkat Cells ; Molecular Weight ; Peptide Mapping ; Quality Control ; Recombinant Fusion Proteins ; biosynthesis ; chemistry

Embryonic hematopoiesis in mammals is characterized by successive temporal and spatial changes. Previous investigations indicate that in vitro differentiation of embryonic stem cells (ES cells) derived from 129 mice can mimic embryonic hematopoiesis to some extent. To investigate the in vitro hematopoietic differentiation capacity of ES cells derived from C57BL/6 mice, the authors initially established the murine ES cell line with standard identification methods employed. Next, two-step culture system was utilized for embryoid bodies formation and the appearance of different hematopoietic precursors was confirmed by CFC assay, cellular chemical staining as well as RT-PCR. The results demonstrated that the ES cell line MES-1 fulfilled the criteria of ES cell line and its progeny after in vitro differentiation included primitive and definitive erythrocyte precursors, mixed colony-forming cells and granulocyte/macrophage colony-forming cells. RT-PCR analysis revealed the molecular consistence of transcription factors and hematopoietic markers with cellular event. In conclusion, MES-1 established from C57BL/6 mice was able to differentiate in vitro to a variety of hematopoietic precursors, thus could partly recapitulate embryonic hematopoiesis.
Animals ; Cell Culture Techniques ; methods ; Cell Differentiation ; genetics ; Cell Line ; Colony-Forming Units Assay ; DNA-Binding Proteins ; genetics ; Embryo, Mammalian ; cytology ; Erythroblasts ; cytology ; metabolism ; Erythroid Precursor Cells ; cytology ; metabolism ; Erythroid-Specific DNA-Binding Factors ; Gene Expression ; Hematopoietic Stem Cells ; cytology ; metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cells ; cytology ; metabolism ; Time Factors ; Transcription Factors ; genetics ; Vascular Endothelial Growth Factor Receptor-2 ; genetics

OBJECTIVETo study the effects of carbon disulfide (CS(2)) on the health of workers, and to provide the basis for the further preventive measures.

METHODSThe occupational health examination of 881 workers exposed to CS(2) in a chemical fiber factory was carried out according to the national technical standard of occupational health surveillance. The time weighted average concentrations (TWA) of CS(2) in the workshops of short silk and long silk exceeded the national standard, The workers in these two workshops served as the high exposure group. The workers of the other workshops were classified as low exposure group, in which TWA met the national standard. All exposed workers were also divided into long-term exposure group and short-term exposure group on the basis of average exposure years (16 years). The statistical analysis was used by t test, χ(2), and fisher exact test.

RESULTSThe symptom rates of numbness, fatigue, dizziness, insomnia and headache were 12.5%, 8.5%, 8.2%, 7.5%, and 7.2% respectively. The abnormal rates of superficial sensation, three fibrillation, achilles tendon reflex, patellar reflex, EMG, hypertension, triglycerides, low-density lipoprotein, and apolipoprotein B were 33.0%, 26.1%, 20.8%, 18.6%, 10.8%, 33.4%, 24.5%, 17.0% and 9.3% respectively. Among the high exposure group, the abnormal detectable rates of dizziness, headache, fatigue, insomnia, numbness, achilles tendon reflex, superficial and deep sensation, EMG, hypertension, ECG, total cholesterol, triglyceride and low density lipoprotein in long-term exposure group were significantly higher than those in short-term exposure group (P < 0.05 or P < 0.01). However, among the low exposure group, the abnormal rates of the insomnia achilles tendon reflex and superficial sensation in long term exposure group were significantly higher than those of short term exposure group (P < 0.05 or P < 0.01). The detected rates of hypertension, triglycerides, low-density lipoprotein, creatinine and uric acid were in males significantly greater than those in females (P < 0.05 or P < 0.01).

CONCLUSIONLong-term exposure to high concentrations of CS(2) could lead to the damage of nervous system, elevate blood pressure and promote the development and progression of hyperlipemia and atherosclerosis. Furthermore, CS(2) had greater effects on blood pressure and lipid in males than in females.

Adult ; Carbon Disulfide ; adverse effects ; Cardiovascular System ; Chemical Industry ; Female ; Health Status ; Humans ; Male ; Middle Aged ; Multiphasic Screening ; Nervous System Diseases ; epidemiology ; Occupational Exposure ; Young Adult

OBJECTIVETo analyze the clinical characteristics of 267 cases with occupational chronic carbon disulfide (CS(2)) poisoning and to provide the basis for revising the items of periodical medical examination of workers occupationally exposed to CS(2).

METHODSThe subjects of present study were 267 patients with mild CS(2) poisoning diagnosed according to "Diagnostic Criteria of Occupational Chronic Carbon Disulfide Poisoning (GBZ4-2002)" from April in 2006 to May in 2010. All patients were from the same chemical fiber factory. When a subject was diagnosed as patient with CS(2) poisoning, who should interview with questionnaire which included the illness and occupational history, symptoms, individual habits. The physical examination, nervous test, cardiovascular test, biochemical test and electromyogram were performed.

RESULTSThe rate of decreased motor conduction velocity was 87.3% (233/267 roots). The highest detection rate of slowing conduction velocity was the common peroneal motor nerve which was 48.6% (138/248 roots) and the second was median motor nerve with delay rate of 37% (155/419 roots). The main symptoms of the patients were neurasthenia, numbness and paresthesia. The rates of abnormal achilles tendon reflex and knee jerk reflex in patients were were 79.4% and 49.8%, respectively. The detected rates of patients with ST-segment changes and hypertension were 19.1% and 27.5%, respectively. The rates of hypertension, systolic pressure and diastolic pressure were 27.3%, 22.5% and 21.1%, respectively. The rates of lactate dehydrogenase (LDH), triglycerides (TG) and low density lipoprotein (LDL) were high. The detected rates of urine acid, indirect bilirubin and total bilirubin in male patients were higher than those in female patients. In addition, the abnormal detected rate of urea nitrogen and indirect bilirubin increased with exposure years.

CONCLUSIONOccupational chronic CS(2) poisoning mainly affects the nervous system, as well as liver and kidney function. Detecting the median and common peroneal motor nerve conduction velocities could be the screening indicators for the peripheral nerve injury induced by CS(2) in the occupational exposure population during the periodical occupational medical examinations.

Adult ; Aged ; Carbon Disulfide ; poisoning ; Chemical Industry ; Female ; Humans ; Kidney ; drug effects ; physiopathology ; Liver ; drug effects ; physiopathology ; Male ; Middle Aged ; Multiphasic Screening ; Nervous System ; drug effects ; physiopathology ; Neural Conduction ; Occupational Exposure

OBJECTIVETo observe the dynamic change in expression of protease-activated receptor 2 (PAR2) during onset and progression of liver fibrosis by using a rat model.

METHODSA cholestatic liver fibrosis model was established in Sprague-Dawley rats (aged 8-9 weeks, body weight 350 - 400 g) by bile duct ligation surgery. Rats receiving a sham operation and unoperated rats served as the negative and normal control groups, respectively. At baseline (pre-surgery) and post-surgery weeks 2, 4, 6, and 8, five rats from each group were sacrificed for whole liver resection. The protein and mRNA expressions of PAR2 and collagen I/III were detected by western blotting and RT-PCR, respectively. Between-group differences were assessed by analysis of variance testing.

RESULTSAt post-surgery week 2, the liver fibrosis group showed higher expression of PAR2 mRNA and protein than either control group. The expression levels of PAR2 continued to rise over time in the liver fibrosis group (peaking at week 8), and were significantly higher than those detected in the control groups (weeks 4-6: P less than 0.05; week 8, P less than 0.05). A similar trend was observed for the expression of collagen I/III.

CONCLUSIONDynamic expression of PAR2 observed in a cholestatic liver fibrosis rat model may indicate a role for this factor in the formation of liver fibrosis.

Animals ; Collagen Type I ; metabolism ; Collagen Type III ; metabolism ; Disease Models, Animal ; Liver ; metabolism ; pathology ; Liver Cirrhosis, Biliary ; metabolism ; pathology ; Male ; Rats ; Rats, Sprague-Dawley ; Receptor, PAR-2 ; metabolism

The hematopoietic system of the mouse arises from extraembryonic mesoderm that migrate through primitive streak to the presumptive yolk sac at day 7.0 of gestation. However, the mechanisms regulating mesoderm commitment to hematopoietic lineages remain poorly understood. Previous studies demonstrated that the development kinetics and growth factor responsiveness of hematopoietic precursors derived from embryonic stem cells (ES cells) is similar to that found in the yolk sac, indicating that the onset of hematopoiesis within the embryoid bodies (EBs) parallels that found in the embryo. Furthermore, in vitro differentiation of ES cells to hematopoietic cells is valuable for establishment of therapeutic clone against a variety of hematological disorders. Despite the identification of multipotential hematopoietic progenitors in EBs, a subset of more primitive progenitors, identical to the high proliferative potential colony-forming cells (HPP-CFC) derived from human and murine hematopoietic tissues, have not been clearly identified regarding particular their replating potential in vitro. HPP-CFC is among the most primitive hematopoietic multipotent precursors cultured in vitro. In this study, our aim was to investigate the in vitro and in vivo hematopoietic capacity of HPP-CFC within the day 12 EBs, rather than the expansion of more committed progenitors. In this study the HPP-CFC could be detected within EBs differentiated for 5 to 14 days of murine ES cells, but the development dynamics of the HPP-CFC differed greatly among distinct serum lots. Qualitatively HPP-CFC is capable of forming secondary colonies. As to our expectation the ES cells-derived HPP-CFC demonstrated similar regeneration capacity to those from yolk sac, giving rise to secondary granulocyte, erythrocyte, macrophage and mast cells, however largely differed from the counterparts of adult bone marrow. In addition, by RT-PCR ES cells-derived HPP-CFC were found to express transcription factors associated closely with stem cell proliferation including SCL, GATA-2 and AML1 as well as various receptors of hematopoietic growth factors such as c-kit, GM-CSF receptor and interleukin 3 receptor et al. Finally, in order to understand the in vivo hematopoietic capacity of the ES cells-derived HPP-CFC, spleen colony-forming unit (CFU-S) assay was performed. Nevertheless, typical CFU-S was not observed after transplantation of the day 12 EB cells or HPP-CFC colonies into lethally irradiated adult murine. In conclusion the HPP-CFC differentiated from murine ES cells displayed robust hematopoietic activity in vitro, however their in vivo reconstitution ability was not detected. The difference between in vitro and in vivo hematopoietic activities of ES cells-derived primitive hematopoietic precursors deserves further investigation.
Animals ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Cell Differentiation ; genetics ; physiology ; Colony-Forming Units Assay ; Core Binding Factor Alpha 2 Subunit ; genetics ; Embryonic Stem Cells ; cytology ; GATA2 Transcription Factor ; genetics ; Hematopoietic Stem Cells ; cytology ; metabolism ; Humans ; Mice ; Proto-Oncogene Proteins ; genetics ; Proto-Oncogene Proteins c-kit ; genetics ; Receptors, Granulocyte-Macrophage Colony-Stimulating Factor ; genetics ; Receptors, Interleukin-3 ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; T-Cell Acute Lymphocytic Leukemia Protein 1

CD40/CD40L interactions play a pivotal role in T cell activation, and take part in many physiologic and pathologic procedures and different levels. In this article, stable CHO transformants secreting human CD40-Ig fusion protein were established through transfection and selection with Lipofectamaine and G418, respectively. In order to obtain great valume of recombinant protein, big batch serum-free cultures of engineered CHO cells were performed in roller-bottle using CHO-II-SFM medium. After cultures, the cell-culture supernatants were harvested, concentrated through ultra-filtration, and finally purified by affinity choromatography with Protein G Sepharose Fast Flow. Human peripheral bloods were collected freshly and seperated with Ficoll, CFU-T was cultured in semi-solid culture system with peripheral blood mononuclear cells (PBMNC). Effect of human CD40-Ig fusion protein on the formation of CFU-T was observed in vitro. The results showed that the yield of human CD40-Ig fusion protein was 30 mg in total 3 liter CHO-II-SFM culture supernatant, and it supposed that the expression level of CD40-Ig in CHO cells was more than 10 micro g/ml. The purity of purified fusion protein is above 95%. Furthermore, compared with human IgG, human CD40-Ig fusion protein significantly inhibited the formation of CFU-T at dose 0.25, 1.0, 4.0, and 10 micro g/ml, it lays a good foundation to evaluate its potential functions in vivo.