Background The in vitro culture of retinal vascular endothelial cells is the foundation of experimental study of retinal vascular disease. Shortage of human donor eyeballs is a main limiting for the laboratory work. The culture method of rat-derived vascular endothelial cells has been established. However, this method is not enough effective because of severer cellullar injury. Objective Present study was to establish a simple and high effective method for the culture of vascular endothelial cells in vitro. Methods The retinas from 5 SPF SD rats was digested by 0. 1% collagenase and cultured with explant culture method. 20% fetal bovine serum, vascular endothelial growth factor ( VEGF) , insulin-transferrin-selenium( ITS) were composed into the endothelial cell culture medium, and enough blowing was performed to get the cells and fragments from retinal tissue. The cellular suspension was prepared and cultured consequently on human fibronectin-coated culture flasks. Cultured vascular endothelial cells were identified by anti-von Willebrand staining factor. Results The cells emerged from the tissue mass,and cells and some tissue fragments attached to the wall after 24 hours of seeding. The cells grew to show the fusiform in 4 days and merged together in 5 to 6 days,and a cell monolayer was seen in the 14th day after culture. The endothelial cells showed the positive response for von Willebrand factor. After passage, the merging-growth statue of the cells was regained in 2 hours after culture. Conclusion Use of retinal pieces and collagenase-digestion can get the vascular endothelial cells with better activity in vitro. The culture method based on highly selective endothelial cell culture medium associated to FN adhesion-promoting is helpful for gaining the purified of endothelial cells.

Chemotherapy-induced neuropathy is a serious clinical problem for patients receiving cancer treatment. The aim of this study was to investigate the potential efficacy of pregabalin in chemotherapy-induced neuropathy in rats. A total of 35 male Sprague-Dawley rats were randomly divided into 5 groups: group 1, naive control; group 2, treated with pregabalin (30 mg/kg p.o., for 8 days); group 3, docetaxel was given by single intravenous infusion at 10 mg/kg; groups 4 and 5, pregabalin at 10 mg/kg and 30 mg/kg respectively was orally administered for 8 days after the docetaxel treatment. On day 8, behavioral test was performed, and substance P and CGRP release in dorsal root ganglion (DRG) and sciatic nerve were analyzed by electron microscope. Our results showed that docetaxel induced mechanical allodynia, mechanical hyperalgesia, heat hypoalgesia, cold allodynia, and sciatic nerve impairment and substance P and CGRP release in DRG. However, oral administration of pregabalin (10 mg/kg and 30 mg/kg) for 8 consecutive days significantly attenuated docetaxel-induced neuropathy by ameliorating heat hypoalgesia, cold allodynia, impairment of sciatic nerve and reducing the release of substance P and CGRP. The findings in the present study reveal that pregabalin may be a potential treatment agent against chemotherapy-induced neuropathy.

Objective To find out iodine nutritional status of pregnant women in the third trimester and the relationship between pregnant women and their neonates thyroid function in a high iodine area.Methods From April to June 2010,210 copies of fasting morning urine and venous blood,and their neonatal umbilical cord blood samples were collected in Haixing Hospital,Cangzhou city,Hebei province.Urinary iodine was determined by arsenic-cerium catalytic spectrophotometry.The levels of free triiodothyronine(FT3),free thyroxine (FT4) and sensitive thyroid-stimulating hormone (sTSH) in serum were measured by chemiluminescence.Results Median urinary iodine of 210 pregnant women(average age 27.69 ± 4.73 years) in the third trimester was 1240.70 μg/L,84.3％(177/210) of them was in excessive iodine nutrition,and only 0.5％(1/210) of them was in adequate iodine nutrition.The prevalence rate of thyroid diseases was 19.5％ (41/210),and the spectrum of diseases were subclinical hypothyroidism(16.2％,34/210),subclinical hyperthyroidism(0.9％,2/210),hypothyroidism(2.4％,5/210) and hyperthyroidism (0,0/210).The number of newborns with sTSH 5 - ＞ 10 mU/L were 104 persons (49.5％); 10 - ＞ 20 mU/L were 44 persons(21.0％),and ≥20 mU/L were 16 persons(7.6％).Of pregnant women suffer from thyroid disease,the ratio(50.0％,24/48 ) of sTSH equal to 10.18 mU/L and ＞ 10 mU/L of their neonates was higher than that of their corresponding non-ill pregnant women(6.78 mU/L,Z =- 2.867,P ＜ 0.05; 22.2％,36/162,x2 =14.000,P ＜ 0.05).There was a positive correlation between neonates' and their mothers' sTSH levels (r =0.278,P ＜ 0.05).There was also a positive correlation between neonates' (sTSH ＞ 10 mU/L) and their mothers' abnormal sTSH levels (r =0.240,P ＜ 0.05).Conclusions Most of the pregnant women in high iodine areas are iodine excess.The level of neonates' sTSH is higher,and it is due to their mothers' abnormal sTSH and suffering from thyroid diseases to some extent.As a result,the monitoring of pregnant women's iodine nutrition and thyroid function and sTSH level of their neonates should be strengthened.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Leukemia, Prolymphocytic, B-Cell ; diagnosis ; Male ; Middle Aged ; Retrospective Studies

OBJECTIVETo evaluate the prognostic factors in treating primary liver cancer (PLC) patients by Pi-strengthening and qi-regulating method (PSQRM), thus providing evidence and optimizing Pi-strengthening and qi-regulating program.

METHODSClinical data of 151 PLC patients treated by PSQRM at Oncology Department, Guangdong Provincial Hospital of Traditional Chinese Medicine from May 2007 to March 2009 were retrospectively analyzed. The univariate analysis was determined to analyze possible prognostic factors. Selected key factors were introduced into the COX proportional hazard model, and multivariate analysis was carried out.

RESULTSThe 1-year survival rate was 21.85%, the median survival time was 6.80 months, and the mean survival time was 8.98 months. The univariate analysis showed that Chinese medicine (CM) syndrome types, clinical symptoms at the initial diagnosis, ascites, tumor types, ratios of foci, portal vein tumor thrombus, intrahepatic metastasis, a-fetoprotein (AFP) levels, total bilirubin classification, albumin classification, Child-Pugh classification, and domestic staging of liver cancer were significant prognostic factors (P < 0.05). The statistic data of multivariate analysis indicated that CM syndrome types, ascites, tumor types, portal vein tumor thrombus, AFP levels, Child-Pugh classification, and domestic staging of liver cancer were independent factors influencing prognosis (P < 0.05).

CONCLUSIONThe prognosis of PLC treated with PSQRM is determined by multiple factors including CM syndrome types, ascites, tumor types, portal vein tumor thrombus, AFP levels, Child-Pugh classification, and domestic staging of liver cancer.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; epidemiology ; therapy ; Female ; Humans ; Liver Neoplasms ; epidemiology ; therapy ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Multivariate Analysis ; Prognosis ; Retrospective Studies ; Survival Rate ; Treatment Outcome

OBJECTIVETo explore the prognostic impact of bone marrow involvement (BMI) and therapy in diffuse large B cell lymphoma (DLBCL).

METHODSThe clinical characteristics and prognosis of 83 DLBCL patients with or without BMI were retrospectively analyzed. The treatment outcome of standard CHOP regimen (CHOP group), intensive-dose regimen (intensive-dose group) and rituximab combined therapy (rituximab group) were compared.

RESULTSThe adverse prognostic factors including LDH elevation, ECOG score > or =2, higher IPI and aaIPI score, B symptom, hepatomegaly, splenomegaly, hemoglobin <110 g/L, platelet <100 x 10(9)/L and serum albumin <35 g/L were more prevalent in DLBCL patients with BMI than in those without BMI. Multivariate analysis showed that BMI was an independent prognostic factor of DLBCL. The 3-year OS and PFS rates in rituximab group were 78.1% and 64.3%, respectively, being statistically higher than that in CHOP group (23.6% and 21.8% respectively, P = 0.000 for both) and in intensive-dose group (33.3% and 25.7% respectively, P = 0.002 and 0.001, respectively). But no difference between the latter two groups (P = 0.411 and 0.694, respectively). For the patients with BMI, the 3-years OS and PFS in rituximab group (57.1% and 57.1%) were statistically higher than that in CHOP group (13.9% and 14.1%) and intensive-dose group (29.5% and 16.8%) (P = 0.029 and 0.012 respectively), respectively and also no difference in the latter two groups (P = 0.226 and 0.376 respectively). In the rituximab group, the 3-years OS and PFS were 86.7% and 67.3% respectively in patients without BMI, being higher than that in patients with BMI (57.1% and 57.1%), but the difference was not statistically significant (P = 0.645 and 0.965 respectively).

CONCLUSIONBMI is a negative independent prognostic factors of DLBCL patients. The rituximab combined chemotherapy can significantly improve the therapeutic effect of the DLBCL, and relieve the negative impact of BMI.

Adolescent ; Adult ; Aged ; Antibodies, Monoclonal, Murine-Derived ; administration & dosage ; Antineoplastic Combined Chemotherapy Protocols ; administration & dosage ; therapeutic use ; Bone Marrow ; pathology ; Child ; Child, Preschool ; Cyclophosphamide ; administration & dosage ; therapeutic use ; Doxorubicin ; administration & dosage ; therapeutic use ; Female ; Humans ; Lymphoma, Large B-Cell, Diffuse ; drug therapy ; pathology ; Male ; Middle Aged ; Neoplasm Invasiveness ; Prednisone ; administration & dosage ; therapeutic use ; Prognosis ; Retrospective Studies ; Rituximab ; Treatment Outcome ; Vincristine ; administration & dosage ; therapeutic use ; Young Adult

OBJECTIVETo standardize the establishment of a stable rat model of orthotopic liver transplantation and surgical technique of two-cuff technique for improving the operation success rate, and compare the effect of graft perfusion via the portal vein and the abdominal aorta on the graft function.

METHODSTotally 100 cases of orthotopic liver transplantation was performed in rats under microscope according to modified Kamada's two cuff technique. The rats were divided into 2 groups with the donor liver perfused through the portal vein with 10 ml cold lactated Ringer's solution, and via the abdominal aorta with 20 ml cold lactated Ringer's solution, respectively. The postoperative function recovery and pathological changes of the liver grafts were evaluated by serum ALT detection and histopathological examination. The operation success rate, 3-month survival rate of the rats and the complications were observed.

RESULTSNo significant differences was noted in the liver function, operation success rate and 3-month survival rate between the 2 groups, and histopathological examination also showed similar findings. The success rates of the two groups were 98% and 96%, with 3-month survival rate of 93.5% (29/31) and 93.3% (28/30) (P>0.05), respectively.

CONCLUSIONThe liver transplantation models with portal vein or abdominal aorta graft perfusion both serve well their respective purposes. Good microsurgical skills, standardized performance and shortened anhepatic period are the keys to improved stability and survival rate and reduced operative complications.

Animals ; Aorta, Abdominal ; surgery ; Liver ; blood supply ; Liver Transplantation ; methods ; mortality ; Male ; Models, Animal ; Portal Vein ; surgery ; Rats ; Rats, Wistar ; Survival Rate

ZM-1 tissue microarrayer designed by our group is manufactured in stainless steel and brass. It features an easier and faster preparation for tissue microarrays. By means of it, a group of biopsy needles are used to punch the donor tissue specimens respectively, and all the needles with the punched specimen cylinders are arranged into the array-board, where small holes have been digged to fit the needles. All the specimen cylinders arraying and the tissue microarray block's shaping are finished simultaneously. ZM-1 tissue microarrayer with a lower cost of manufacture, is capable of preparing the tissue microarrays conveniently, efficiently and quality-controllably.
Equipment Design ; Tissue Array Analysis ; instrumentation

The ZM-1 tissue microarrayer designed by our groups is manufactured in stainless steel and brass and contains many features that make TMA (tissue microarray) paraffin blocks construction faster and more convenient. By means of ZM-1 tissue microarrayer, biopsy needles are used to punch the donor tissue specimens respectively. All the needles with the punched specimen cylinders are arrayed into the array-board, with an array of small holes dug to fit the needles. All the specimen cylinders arraying and the TMA paraffin block shaping are finished in only one step so that the specimen cylinders and the paraffin of the TMA block can very easily be incorporated and the recipient paraffin blocks need not be made in advance, and the paraffin used is the same as that for conventional pathology purpose. ZM-1 tissue microarrayer is easy to be manufactured, does not need any precision location system, and so is much cheaper than the currently used instrument. Our method's relatively cheap and simple ZM-1 tissue microarrayer technique of constructing TMA paraffin block may facilitate popularization of the TMA technology.
Biopsy, Needle ; instrumentation ; Equipment Design ; Female ; Humans ; Immunohistochemistry ; Male ; Neoplasms ; enzymology ; Paraffin ; Tissue Array Analysis ; instrumentation

Low birth weight (LBW) and preterm birth (PB) are associated with newborn mortality and diseases in adulthood.We explored factors related to LBW and PB by conducting a population-based case-control study from January 2011 to December 2013 in Wuhan,China.A total of 337 LBW newborn babies,472 PB babies,and 708 babies with normal birth weights and born from term pregnancies were included in this study.Information of newborns and their parents was collected by trained investigators using questionnaires and referring to medical records.Univariate and logistic regression analyses with the stepwise selection method were used to determine the associations of related factors with LBW and PB.Results showed that maternal hypertension (OR=6.78,95％ CI:2.27-20.29,P=0.001),maternal high-risk pregnancy (OR=1.53,95％ CI:1.06-2.21,P=0.022),and maternal fruit intake ≥300 g per day during the first trimester (OR=1.70,95％ CI:1.17-2.45,P=0.005) were associated with LBW.BMI ≥24 kg/m2 of mother prior to delivery (OR=0.48,95％ CI:0.32-0.74,P=0.001) and gestation ≥37 weeks (OR=0.01,95％ CI:0.00-0.02,P＜0.034) were protective factors for LBW.Maternal hypertension (OR=3.36,95％ CI:1.26-8.98,P=0.016),maternal high-risk pregnancy (OR=4.38,95％ CI:3.26-5.88,P＜0.001),maternal meal intake of only twice per day (OR=1.88,95％ CI:1.10-3.20,P=0.021),and mother liking food with lots of aginomoto and salt (OR=1.60,95％ CI:1.02-2.51,P=0.040) were risk factors for PB.BMI ≥24 kg/m2 of mother prior to delivery (OR=0.66,95％ CI:0.47-0.93,P=0.018),distance of house from road ≥36 meters (OR=0.72,95％ CI:0.53-0.97,P=0.028),and living in rural area (OR=0.60,95％ CI:0.37-0.99,P=0.047) were protective factors for PB.Our study demonstrated some risk factors and protective factors for LBW and PB,and provided valuable information for the prevention of the conditions among newborns.