2.Comparison of immunohistochemistry and fluorescence in situ hybridization in detecting c-erbB-2 expression in breast cancer
Wei SHENG ; Xiangming CHE ; Tao SHAN ; Lin FAN ; Meng LI ; Qian ZHANG ; Xitao GAO
Journal of Xi'an Jiaotong University(Medical Sciences) 2010;31(2):208-211
Objective To compare the consistency of immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) in detecting c-erbB-2 status in breast cancer tissues. Methods A total of 50 breast cancer paraffin embedded samples were selected, of which there were 10 cases of c-erbB-2 protein expression (+), 20 cases of (++) and 20 cases of (+++). FISH was used to assess the amplification of c-erbB-2 gene, and SPSS 13.0 software was employed to analyze the difference and consistency between the two methods. Results IHC and FISH methods had a good consistency when detecting c-erbB-2 (+) and (+++) expressions in breast cancer tissues, with the coincidence rate of 89.2%. However, when IHC was used to test c-erbB-2 (++), the result of FISH was quite different, with the coincidence rate of only 35.3%. Conclusion IHC is a preliminary method to detect c-erbB-2 status in breast cancer. IHC and FISH methods have a good consistency in detecting c-erbB-2 (+) and (+++) status in breast cancer tissues. As detection of c-erbB-2 (++) with IHC has a different result from FISH, such patients should receive FISH confirmation for herceptin therapy.
3.Effect of hypoxia and hyperglycemia on retinal pigment epithelial cells co-cultured with human mesenchymal stem cells
Jie-Jing, YAN ; Hai-Yan, WANG ; Yu-Sheng, WANG ; Fan, GAO ; Na, LI ; Peng, ZHANG
International Eye Science 2015;(4):584-587
AIM: To evaluate cell proliferation, apoptosis and migration of human retinal pigment epithelial cells ( RPE) when co - cultured with human marrow mesenchymal stem cells ( hMSCs ) in condition of hypoxia and hyperglycemia so as to explore possible mechanisms of diabetes aggravating choroidal neovascularization ( CNV) preliminarily.
METHODS:Both hMSCs and RPE cells were co-cultured in a transwell system. The experiment was divided into four groups: 21% O2 with 5. 56mmol/L glucose ( control group, A ), 21% O2 with 30mmol/L glucose ( hyperglycemia and normoxia group, B ) , 5% O2 with 5.56mmol/L glucose ( normoglycemia and hypoxia group, C ) and 5% O2 with 30mmol/L glucose ( hyperglycemia and hypoxia group, D) . Cell Counting Kit-8 (CCK-8) was used to detect the proliferation of RPE cells in each group at 12, 24 and 48h respectively. Flow cytometry was performed to observe apoptosis of RPE cells at 24h. Additionally, we assessed migration
capabilities of RPE via transwell assay under the condition of hyperglycemia and hypoxia by co-culturing of hMSCs.RESULTS:In this co-culturing system, at 12, 24 and 48h, group B (1. 61±0. 41, 1. 80±0. 34;1. 91±0. 35), C (1.34±0. 46, 1. 94±0. 40, 2. 14±0. 41) and D (1. 98±0. 47, 2.26±0.42, 2. 55±0. 40) showed significantly higher proliferation rate than group A (0. 92±0. 45, 1. 27±0. 32, 1.59±0. 41, P<0. 05). The migration capabilities of RPE in group B (149. 5±9. 19), C (140±9. 90) and D (170. 5±7. 78) increased dramatically compared with group A ( 114. 5±7.78, P<0.05) at 24h, whereas there was no significant difference of apoptosis ratio among four groups (P>0. 05).
CONCLUSION:By coexistence with hMSCs, the synergy of hyperglycemia and hypoxia can improve migration and proliferation of RPE cells, and have no effect on apoptosis of RPE cells within short period.
4.Simultaneous Determination of 6 Alkaloids in Coptis teeta by HPLC
Zheng PAN ; Yunling GAO ; Sheng JIANG ; Gang FAN ; Yi ZHANG ; Weiguo CAO
China Pharmacy 2017;28(24):3408-3411
OBJECTIVE:To develop a method for simultaneous determination of 6 alkaloids in Coptis teeta.METHODS:The determination was performed on XtimateTM C18 with mobile phase consisted of 30 mmol/L ammonium bicarbonate [containing 0.1% triethylamine and 0.7% ammonia-acetonitrile (gradient elution)] at the flow rate of 1.0 mL/min.The detection wavelength was set at 270 nm,the column temperature was 30 ℃,and the sample size was 10 μL.RESULTS:The linear ranges of jateorrhizine,columbamine,epiberberine,coptisine,palmatine hydrochloride and berberine hydrochloride were 0.85-16.96 mg/L (r=0.999 9),1.25-24.96 mg/L(r=0.999 8),2.05-40.96 mg/L(r=0.999 9),3.65-72.96 mg/L(r=0.999 9),2.88-57.60 mg/L(r=0.999 9) and 13.25-264.96 mg/L(r=0.999 9),respectively.RSDs of precision,stability and reproducibility tests were all lower than 3.0%.Recoveries were 97.14%-102.14% (RSD=1.93%,n=6),97.00%-102.00% (RSD=2.06%,n=6),98.18%-101.82 % (RSD=1.79%,n=6),96.15%-101.28% (RSD=2.06%,n=6),96.88%-101.88% (RSD=1.87%,n=6),99.31%-103.76% (RSD=1.89%,n=6),respectively.CONCLUSIONS:The method is simple,accurate,stability and reproducible,and can be used for simultaneous determination of 6 alkaloids in Coptis teeta.
5.Influence of posterior intrascleral iron foreign body on rabbit retina and opticus
Yong-sheng, YOU ; Li-jun, SHEN ; Hua, QIN ; Chao, GAO ; Lan, WANG ; Yang, LIU ; Wei, FAN ; Yi-qi, CHEN
Chinese Journal of Experimental Ophthalmology 2011;29(9):809-824
BackgroundIron-containing foreign body trapped in the eyeball wall without affecting the opticus occurs occasionally in clinic. Operation always is performed in an attempt to avoid the deposition of rust in different tissues of the eye-balls. However,a few animal experimental studies showed that a small foreign body does not affect the retina and opticus in the period of three months. The question of whether surgery needs to be carried out is worth discussion. ObjectiveThe aim of this study was to evaluate the influence of posterior intrascleral iron foreign body on the rabbit retina and opticus. MethodsTwelve healthy adult Japan flap-eared white rabbits were randomly divided into two groups. Medium carbon iron with rust or without rust( size of 2. 0 mm × 1. 0 mm×0. 2 mm) were implanted into the posterior sclera of the left eye to create the animal model with iron foreign body in the eyeball wall. The cornea, anterior chamber, crystalline lens, vitreous and fundus of the rabbits were observed under a slit lamp microscope 1weekbeforeoperationand 1week, 2weeks, 1monthand 3months after operation.Flash electroretinogram(F-ERG) and flash visual evoked potential (F-VEP) were recorded at the time points mentioned above. All the rabbits were sacrificed and the eye balls were extracted at the end of the experiment, and the position of the iron foreign body was determined. The histopathological examinations of the retina and opticus were performed under the light microscope. This experiment complied with the Regulations for the Administration of Affair Concerning Experimental Animals by State Science and Technology Commission. ResultsThere were no statistical differences for the a-wave amplitude of F-ERG among different time points( F =1. 885,P =0. 129 ) and different treatment groups ( F =1. 188, P =0. 340 ), as with the ERG b-wave amplitude ( time: F =2. 73, P =0. 064 ; group : F =1. 114, P =0. 367). The differences in the latencies of F-VEP N1-wave were insignificant among the different time points( F =1. 605, P =0. 263 ) as well as various groups ( F=1. 556, P =0.314 ), and those of F-VEP P1 -wave were not evidently changed ( time: F =2. 329, P =0. 092 ; group : F =2. 186, P =0. 103 ). No correlations were seen between the time factor and grouping factor ( P > 0. 05 ). There was no apparent siderosis bulbi change during the follow-up duration. No morphological abnormality in the retina and optical nerve was found under the light microscope. At the end of the experiment,intrascleral iron foreign body was wrapped by surrounding tissue in a stable condition. Conclusions The small posterior intrascleral iron foreign body, whether it is oxidized or not, does not produce distinctive functional or pathological damage on retina and opticus in the short term.
6.An experimental study on place reaction of support drainage tube in cholecystojejunostomy
Zuosheng FAN ; Shilin SUN ; Jie LI ; Changcheng QIU ; Hongwei ZHAO ; Tao GAO ; Sheng WANG ; Bo ZHANG ; Kaifu LI ; Yanmin ZHANG
Chinese Journal of Hepatobiliary Surgery 2010;16(11):845-847
Objective To investigate the place reaction of support drainage tube in cholecystojejunostomy and its pathological changes.Methods Eight pigs were randomized into 2 groups.The tube-placing cholecystojeunostomy(n=2)or choledochocholedochostomy(n=2)and non-tube-placing cholecystojejunostomy(n=2)or choledochocholedochostomy(n=2)were performed respectively.All the 8 pigs were sarcrificed 3 months after the operation for pathological examination.Results Gross anatomy catheter group showed anastomotic inflammatory hyperplasia, wall hardening hyperplasia,luminal narrow contracture and stone formation.Microscopic biopsy showed bile duct mucosa in interstitial lymphocytes, plasma cells and other chronic inflammatory cell infiltration, a large number of fibrous tissue hyperplasia.However, these features were not found in pigs without placing the tube.Conclusion The tube can not prevent the stenosis.It might promote the formation of stricture since it can stimulate inflammatory hyperplasia and tissue hardening.
7.Identification of a novel mutation IVS2-2A-->C of SEDL gene in a Chinese family with X-linked spondyloepiphyseal dysplasia tarda.
Chao GAO ; Qiang LUO ; Huai-li WANG ; Xiao-qun GAO ; Qing-tang FAN ; Hua WANG ; Guang-yao SHENG ; Jian-hua ZHOU ; Tie-zheng GAO
Chinese Journal of Medical Genetics 2003;20(1):15-18
OBJECTIVETo identify the mutation of spondyloepiphyseal dysplasia tarda (SEDL) gene in a large Chinese family with X-linked spondyloepiphyseal dysplasia tarda and to make a discussion on the pathogenesis of SEDL at the molecular level.
METHODSIn two patients, four exons comprising the SEDL open reading frame as well as their exon/intron boundaries were analyzed by bi-directional direct sequencing of PCR products. The sequencing results were compared against the normal sequences in GenBank to find the mutation. Then the mutation was identified in other members of the family.
RESULTSA nucleotide substitution of the splice acceptor in SEDL intron 2, IVS2 -2A-->C,was detected in two affected individuals (IV(15) V(3)) in the Chinese family with SEDL, but no sequence change occurring on exons 3-6 was detected. The transversion was also identified in four heterozygous carriers. The mutation was not found in two unaffected male individuals and fifteen normal controls. Furthermore, four potential carriers were identified in the family.
CONCLUSIONThe mutation IVS2 -2A-->C of SEDL gene was firstly determined in the world. The change of the splice acceptor in SEDL intron 2 may cause skipping of exon 3 which is responsible for the disease. Molecular diagnosis can be made by detecting the mutation.
Alternative Splicing ; genetics ; Base Sequence ; Carrier Proteins ; genetics ; China ; Chromosomes, Human, X ; genetics ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Family Health ; Female ; Genetic Linkage ; Humans ; Male ; Membrane Transport Proteins ; Mutation ; Osteochondrodysplasias ; genetics ; pathology ; Pedigree ; Transcription Factors
8.Clinical characteristics of hepatic veno-occlusive disease in 6 children with hematologic neoplasm treated with 6-thioguanine.
Hong-sheng WANG ; Yi-jin GAO ; Jun LI ; Feng-juan LU ; Hui MIAO ; Xiao-wen QIAN ; Xiao-fan ZHU
Chinese Journal of Pediatrics 2010;48(9):708-710
OBJECTIVETo improve the treatment of drug related childhood hepatic veno-occlusive disease (HVOD), clinical characteristics of 6 children with hematologic neoplasm from 2 hospitals of China Children's Leukemia Group (CCLG) treated with 6-thioguanine (6-TG) complicated with HVOD were analyzed.
METHODAll the drug related HVOD patients were treated with CCLG acute lymphoblastic leukemia (ALL)-2008 protocol. They were from Children's Hospital of Fudan University and Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College from April 2008 to April 2009. The diagnosis was made according to the modified Seattle criteria and Baltimore criteria, including 2 or 3 of the following clinical features: hepatomegaly and upper right abdominal pain, jaundice (bilirubin ≥ 35 µmol/L), ascites or confirmed by pathology. The 6 HVOD patients' clinical manifestations, laboratory finding, imageologic and pathologic data were collected and analyzed.
RESULTOf the 6 patients, 2 were males and 4 females. Mean age of the 6 patients was 3.89 years (range from 3 years 1 month to 4 years 11 months). The original disease was acute lymphoblastic leukemia. HVOD occurred during chemotherapy protocols of CAM (CTX + Ara-C + 6-TG) or maintenance period (MTX + 6-TG). Most of 6 HVOD patients presented with pain in liver area, hepatomegaly on imaging, elevated aminotransferase and bilirubin (often ≥ 35 µmol/L), hydroperitonia was common, one with pleural fluid, illegible hepatic veins. All the patients recovered after being treated with hepatoprotective, jaundice-relieving and supportive therapeutics, some patients were treated with low molecular weight heparin. The prognoses were good.
CONCLUSIONHVOD was a serious complication of chemotherapy with 6-TG. Hepatoprotective and jaundice-relieving and low molecular weight heparin could improve the prognosis.
Antineoplastic Agents ; therapeutic use ; Child, Preschool ; Female ; Hepatic Veno-Occlusive Disease ; drug therapy ; Humans ; Leukemia ; therapy ; Male ; Thioguanine ; therapeutic use
9.Characteristics of treating dislocation and fracture of sacroiliac joint through anterior and posterior approches.
Xiao-Hai FAN ; Ping ZHEN ; Ming-xuan GAO ; Jian-jun WANG ; Yun XUE ; Meng LI ; Xiao-wen DENG ; Xu-sheng LI
China Journal of Orthopaedics and Traumatology 2013;26(12):1048-1051
OBJECTIVETo investigate characteristics of treating dislocation and fracture of sacroiliac joint through anterior and posterior approaches.
METHODSBetween January 2006 and September 2012, 39 patients with dislocations and fractures of sacroiliac joint were treated with operation. There were 28 males and 11 females, aged 12 to 64 (mean, 41.3) years old. Seventeen cases were chosen through anterior approach, 13 cases through posterior and 9 cases were combined with anterior and posterior approaches. The anterior approach were made with supine position, performed open reduction, and two plates with 4 holes were used as internal fixation for arthrodesis of sacroiliac joint after reduction; while posterior approach prone position, close or open reduction was performed, and one or two partial thread lag screws, which with 7.3 mm in diameter and 60 to 75 mm in length, were penetrated via sacroiliac joint for fixation under X-ray. Postoperative complications were observed, Matta and Majeed scoring were used to evaluated currative effects.
RESULTSAll patients were followed up from 6 to 36 months. The patient could sit on the bed 2 to 4 weeks after surgery and walk with a crane 6 weeks' later. No breakage or loosening of screw occurred during follow-up. In all but one case with old sacral iliac fractures reducted poor, others obtained good opposition. According to Matta's criteria, 30 cases got excellent results, 8 good and 1 fair. According to Majeed's funtional standard, 14 patients got excellent results, 20 good 4 fair and 1 poor.
CONCLUSIONTreating dislocation and fracture of sacroiliac joint through anterior and posterior approaches can obtain good effects. However, anatomic location, fracture types, degree of displacement, the rate of preoperational reduction, vulnerability assessment of operation and fixation strength should be considered.
Adolescent ; Adult ; Bone Screws ; Child ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; Fractures, Bone ; surgery ; Humans ; Internal Fixators ; Joint Dislocations ; surgery ; Male ; Middle Aged ; Sacroiliac Joint ; injuries ; surgery ; Young Adult
10.Protective and therapeutic effect of apelin on chronic hypoxic pulmonary hypertension in rats.
Xiao-fang FAN ; Qing WANG ; Sun-zhong MAO ; Liang-gang HU ; Lin HONG ; Li-xian TIAN ; Yu-qi GAO ; Yong-sheng GONG
Chinese Journal of Applied Physiology 2010;26(1):9-12
OBJECTIVETo study the role of apelin in the prevention of pulmonary hypertension induced by hypoxia in rats.
METHODSThe animal model of hypoxic pulmonary hypertension was established by exposing the rats to isobaric hypoxic chamber for 4 weeks (8 h/d, 6 d/ w). Forty male Sprague-Dawley rats were randomly divided into control group (NC), hypoxic group(HH), hypoxic with low-dose apelin (5 nmol/(kg x d) group(LA) and high-dose apelin (10 nmol/(kg x d) (HA). [pGlu]apelin-13 was administered into the rats of apelin groups by mini-osmotic pump subcutaneously. The mean pulmonary arterial pressure(mPAP) and the mean carotid arterial pressure (mCAP) were measured by either right or left cardiac catheterization, and the weight ratio of right ventricule/left ventricule plus septum (RV/(LV + S)) were calculated. The Masson's trichrome stained lung specimens were examined by light microscope to examine the vessel wall area/total area (WA/TA), vessel cavity area/total area (CA/TA) and media thickness of pulmonary arterioles (PAMT). Meanwhile, the lung homogenates were assayed for the activity of supeeroxide dismutase (SOD) and the content of malondialdehyde (MDA).
RESULTS(1) mPAP and RV/(LV + S) of HH group were significantly higher than those of NC group. mPAP of LA and HA groups were lower than those of HH group. The RV/(LV + S) of HA group was significantly lower than that of HH group, but there was no significant difference between HH group and LA group. (2) Masson's trichrome staining revealed that WA/TA and PAMT of HH group were higher than those of NC group. Administration of apelin significantly eliminated WA/TA and PAMT in LA and HA groups. (3) CA/TA of HH group was lower than that of NC group. Administration of apelin significantly elevated CA/TA in LA and HA groups. (4) The activity of SOD and content of MDA in HH group was, respectively, lower and higher than those in NC group. Apelin treatment increased the activity of SOD in LA and HA groups while decreased the content of MDA.
CONCLUSIONSApelin could play an important role in treatment of hypoxic pulmonary hypertension of rats and the mechanisms of protection were associated with vasodilation of pulmonary artery and inhibition of oxidative stress.
Animals ; Cardiotonic Agents ; pharmacology ; therapeutic use ; Hypertension, Pulmonary ; etiology ; physiopathology ; prevention & control ; Hypoxia ; complications ; physiopathology ; Intercellular Signaling Peptides and Proteins ; pharmacology ; therapeutic use ; Male ; Oxidative Stress ; drug effects ; Pulmonary Artery ; physiopathology ; Rats ; Rats, Sprague-Dawley ; Vasodilation ; drug effects