1.Preparation of monoclonal antibodies against human mesenchymal stem cells.
Zhi-xiong LIAO ; Jian-gen SHEN ; Xiao-yu LAI
Journal of Zhejiang University. Medical sciences 2006;35(5):517-522
OBJECTIVETo prepare monoclonal antibodies (McAbs) against human mesenchymal stem cells (hMSCs) and to study their biological characteristics.
METHODSBALB/C mice were immunized with pooled hMSCs. McAbs were prepared by hybridoma technique and their biological characteristics were analyzed by indirect immunofluorescence, immunohistochemistry and flow cytometry.
RESULTFive hybridoma cell lines were successfully established, which secret McAbs specifically against hMSCs. Investigations showed that all these McAb reacted only to hMSCs and had no cross-reaction to other human cells, the relative affinities of 5 McAbs were 1x10(6) (ZUB1), 1x10(5) (ZUB4), 1x10(6) (ZUC3), ZUE12 (1x10(5)) and 1x10(5) (ZUF10), respectively. Isotype analysis showed that ZUB1, ZUE12, ZUF10 against the same isotype, while ZUC3, ZUB4 against other two different isotypes alone. Flow cytometric analysis showed that the positive expression rate of cultured hMSCs was 87.39% (ZUB1), 88.07% (ZUB4), 88.12% (ZUC3), 69.89% (ZUE12) and 83.67% (ZUF10).
CONCLUSIONThe prepared five McAbs can specifically react against hMSCs, which can be used for selection and study of hMCSs.
Animals ; Antibodies, Monoclonal ; biosynthesis ; Antibody Specificity ; Bone Marrow Cells ; cytology ; immunology ; Fluorescent Antibody Technique ; methods ; HL-60 Cells ; Humans ; Hybridomas ; secretion ; Immunoglobulin G ; immunology ; Immunoglobulin M ; immunology ; Mesenchymal Stromal Cells ; cytology ; immunology ; Mice ; Mice, Inbred BALB C ; Rats
2.Diagnostic values of ultrasound and (18)F-fluoro-2-deoxy-D-glucose-positron emission tomography/computerized tomography for patients with suspected thyroid carcinoma and lymph node metastasis.
Xing-jian LAI ; Bo ZHANG ; Yu-xin JIANG ; Qing DAI ; Rui-na ZHAO
Acta Academiae Medicinae Sinicae 2013;35(4):393-397
OBJECTIVETo evaluate the diagnostic values of ultrasound (US) and (18)F-fluoro-2-deoxy-D-glucose-positron emission tomography (FDG PET)/computerized tomography (CT) in diagnosing suspected thyroid carcinoma and lymph node metastasis.
METHODSThe clinical data of 28 patients who had undergone total or subtotal thyroidectomy with or without neck dissection from December 2011 to December 2012 in PUMC Hospital and had undergone US and FDG PET/CT before surgery were retrospectively analyzed. In each patient, US and FDG PET/CT images were retrospectively reviewed to determine the presence of carcinoma with or without loco-regional metastasis by level-by-level analysis. The potential correlation between imaging results and histopathology were analyzed.
RESULTSThere were 11 benign lesions,15 papillary carcinomas, one follicular carcinoma, and one medullary carcinoma. For thyroid carcinoma,the sensitivity and specificity were 88.2% and 63.6% for US and 76.5% and 54.5% for FDG PET/CT(P>0.05). For lymph node metastasis, the sensitivity was 68.0% for US and 60.0% for FDG PET/CT (P>0.05), and the specificity was 96.7% for US and FDG PET/CT.FDG PET/CT could provide more diagnostic information than US for patients with level 2 or 5 metastasis.
CONCLUSIONSCombination of US and FDG PET/CT is typically not needed for differentiating thyroid lesions.However, for patients with suspected lymph node metastasis of infrequently involved levels, the combination of US and FDG PET/CT may be a good choice.
Adolescent ; Adult ; Aged ; Female ; Fluorodeoxyglucose F18 ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; diagnosis ; Male ; Middle Aged ; Positron-Emission Tomography ; Retrospective Studies ; Sensitivity and Specificity ; Thyroid Neoplasms ; diagnostic imaging ; Tomography, X-Ray Computed ; Ultrasonography ; Young Adult
3.Mechanism of telomere mainteance in human bone marrow mesenchymal stem cells
Jingyuan LI ; Jianping LAN ; Yanmin ZHAO ; Xiaoyu LAI ; Yi LUO ; Jie SUN ; Jian YU ; Yuanyuan ZHU ; Fenfang ZENG ; He HUANG
Chinese Journal of Pathophysiology 1999;0(09):-
AIM:To study the telomere maintenance mechanism in mesenchymal stem calls(MSCs).METHODS:MSCs were isolated from healthy human bone marrow by their adherence to plastic and then were checked with CD14-FITC,CD45-FITC,CD44-FITC,HLA-DR-FITC,CD34-PE,CD29-PE and CD166-PE.Telomere length and ECTR DNA in MSCs were detected by Southern blotting.The localization of TRF1 and promyelocytic leukemia(PML)in MSCs were detected with immunofluorescence staining.TRAP protocol was performed to detect the telomerase activity in MSCs and MSCs-derived adipocytes.Western blotting and TRAP protocol were applied to measure telomerase activity of MSCs,which were synchronized by serum starvation and aphidicolin treatment.RESULTS:The telomere in length seemed shorter and relatively more homogeneous in MSCs and HeLa cells than that in WI-38-2RA cells.TRF1 did not concide with PML nuclear body in MSCs and HeLa cells while it exclusively did in WI-38-2RA cells.ECTR DNA was negative in MSCs and HeLa cells but positive in WI-38-2RA cells.Telomerase was negative in MSCs but it was positive in MSCs-derived adipocytes detected by TRAP.Moreover,a cell cycle-dependent expression profile of telomerase was found in MSCs when they were synchronized by serum starvation and aphidicolin treatment.Untreated MSCs expressed very low level of telomerase probed by Western blotting with 2C4 mAb,but the telomerase level had significantly increased when these cells were trapped in S phase.CONCLUSION:The telomere of MSCs is maintained by telomerase pathway instead of alternative lengthing of telomere(ALT)and the level of telomerase expression is associated with cell cycle stage.[
4.Studies on hypokalemia induced by trimethyltin chloride.
Xiao-Jiang TANG ; Guan-Chao LAI ; Jian-Xun HUANG ; Lai-Yu LI ; Ying-Yu DENG ; Fei YUE ; Qing ZHANG
Biomedical and Environmental Sciences 2002;15(1):16-24
OBJECTIVESTo determine the possible relationship between plasma potassium concentration and severity of acute trimethyltin chloride (TMT) poisoning and to assess the mechanism of TMT induced hypokalemia.
METHODSSD rats were treated with various dosages of TMT (i.p.). All the indices were measured and analysed for determining their possible relations with plasma K+.
RESULTSWith increase of dosage, the plasma K+ level dropped rapidly, and deaths appeared more quickly. The LD50 of TMT (i.p.) was 14.7 mg/kgbw. In the low dosage group (10 mg/kgbw), the plasma K+ level dropped slowly with the lowest dosage on day 6 (4.85 mmol/L). It rose again on day 11 (5.06 mmol/L), and recovered on day 28. The poisoning signs corresponded with decline of the span of K+ level. The plasma Na+ level dropped half an hour after TMT treatment, but recovered 24 h later. In the high dosage group (46.4 mg/kgbw), the levels of plasma K+ and Na+ fell rapidly within half an hour (P < 0.05), the intracellular potassium concentration of RBC did not decrease obviously (P > 0.05), the activities of Na(+)-K(+)-ATPase and Mg(2+)-ATPase in RBC membrane were depressed remarkably (P < 0.01, P < 0.05, respectively), the plasma aldosterone concentrations rose as high as tenfold (P < 0.01), the arterial blood pH fell from 7.434 to 7.258 (P < 0.01), pCO2 was raised from 29.62 to 45.33 mmHg (P < 0.01). In the 24 h urine test, when rats were treated with TMT (21.5 mg/kgbw, i.p.), urine volume, urinary potassium, sodium and chloride increased significantly in comparison with those in the controls (P < 0.01).
CONCLUSIONTMT could induce hypokalemia in SD rats. The available evidence suggests that TMT can induce acute renal leakage of potassium. At the same time, a significant rise of plasma aldosterone may play an important role in promoting potassium leakage from kidney to result in severe hypokalemia with inhaling acid-base abnormalities produced, which aggravate the poisoning symptoms. In the end the rats would die of respiratory failure.
Animals ; Female ; Hypokalemia ; chemically induced ; veterinary ; Injections, Intraperitoneal ; Kidney Diseases ; chemically induced ; veterinary ; Lethal Dose 50 ; Male ; Rats ; Rats, Sprague-Dawley ; Severity of Illness Index ; Trimethyltin Compounds ; pharmacology ; poisoning
5.Comparison of different diameter hollow screw for the treatment of femoral neck fractures.
Yu-liang LOU ; Jian-jun HONG ; Ke-he YU ; Xiao-lang LU ; Yi-jing ZHENG ; Hong-yan LAI ; Xi-wen SHAO
China Journal of Orthopaedics and Traumatology 2015;28(9):792-795
OBJECTIVETo evaluate and compare the outcome of two kinds of diameter hollow screws for the treatment of femoral neck fractures.
METHODSFrom June 2008 to June 2013, 117 patients with femoral neck fractures were treated by closed reduction and hollow screws fixation. Among them,48 patients were fixed by 6.5 mm screw including 30 males and 18 females with an average age of (45.61 ± 11.99) years old ranging from 19 to 60 years old, involving 17 cases in Garden I/II and 31 cases in Garden III/IV; 69 patients were fixed by 8.0 mm screw including 40 males and 29 females with an average age of (45.17 ± 9.95) years old ranging from 18 to 60 years old, involving 31 cases in Garden I/II and 38 cases in Garden III/IV. The general information, operative time, hospital stay time, reduction quality, diameter of femoral head and neck, fracture healing time, the rate of fracture healing, postoperative complications were recorded and evaluated. Harris scoring was used to evaluate the hip joint function.
RESULTSAll patients were followed up for 19.6 months (18 to 24 months). The difference of operative time, duration of hospitalization, quality of reduction were not statistically significant (P > 0.05). There was no difference between two groups about the average diameter of the femoral head and neck, the fracture healing time, the rate of healing and the postoperative complications (P > 0.05). There were no difference between two groups about Harris scale. There were significant difference between Garden III/IV and I /II (P > 0.05).
CONCLUSIONClosed reduction and internal fixation with hollow screw in treating the young adult patients with femoral neck fracture is the first choice, both different diameters hollow screws could meet the requirements of fixation of femoral neck fracture, and not affect on fracture healing time and postoperative complications.
Adult ; Bone Screws ; Female ; Femoral Neck Fractures ; surgery ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged
6.Guinea pig maximization test for trichloroethylene and its metabolites.
Xiao-Jiang TANG ; Lai-Yu LI ; Jian-Xun HUANG ; Ying-Yu DENG
Biomedical and Environmental Sciences 2002;15(2):113-118
OBJECTIVESTo study the contact allergenic activities of trichloroethylene (TCE) and its three metabolites trichloroacetic acid, trichloroethanol and chloral hydrate.
METHODSA modified guinea pig maximization test (GPMT) was adopted. The skin sensitization (edema and erythema) was observed in trichloroethylene, trichloroacetic acid, trichloroethanol, chloral hydrate and 2,4-dinitrochlorobenzene.
RESULTSThe allergenic rate of TCE, trichloroacetic acid and 2,4-dinitrochlorobenzene was 71.4%, 58.3% and 100.0% respectively, and that of trichloroethanol and chloral hydrate was 0%. The mean response score of TCE, trichloroacetic acid and 2,4-dinitrochlorobenzene was 2.3, 1.1, 6.0 respectively. The histopathological analysis also showed an induction of allergenic transformation in guinea pig skin by both TCE and trichloroacetic acid.
CONCLUSIONTCE appears to be a strong allergen while trichloroacetic acid a moderate one. On the other hand, both trichloroethanol and chloral hydrate are weak sensitization potentials. Immunologic reaction induced by TCE might be postulated as the pathological process of this illness. Consequently, it is suggested that in the mechanism of Occupational Dermatitis Medicamentose-Like (ODML) induced by TCE, the chemical itself might be the main cause of allergy. As one of its metabolic products, trichloroacetic acid might be a subordinate factor.
Allergens ; toxicity ; Animals ; Chloral Hydrate ; toxicity ; Dermatitis, Allergic Contact ; etiology ; immunology ; Dermatitis, Irritant ; etiology ; immunology ; Ethylene Chlorohydrin ; analogs & derivatives ; toxicity ; Guinea Pigs ; Skin ; drug effects ; immunology ; Toxicity Tests ; Trichloroacetic Acid ; toxicity ; Trichloroethylene ; metabolism ; toxicity
7.Risk factors of recurrence after transurethral resection of the prostate due to benign prostate hyperplasia.
Jian-yu LAI ; Ling-min SONG ; Yu-chun ZHU ; Qiang WEI
Chinese Journal of Surgery 2010;48(2):103-107
OBJECTIVESTo review the clinic data of benign prostate hyperplasia (BPH) cases, and to find the risk factors of recurrence in post transurethral resection of the prostate (TURP) patients.
METHODSFrom November 2002 to November 2007, 1471 cases were reviewed, including 41 patients of recurrence after TURP. Record the data include onset age, course of disease, age of surgery, LUTS, PSA, blood serum creatinine, size of the prostate in transabdomen ultrasonography, data of urodynamic examination, weight of resected tissue, persistence time of the resection, length of stay, score of inflammation of the pathologic sample, experience of the operator, routine blood examination, routine urine examination and so on. Statistical analysis was performed using univariate and multivariate unconditional Logistic regression analysis for evaluation of the data.
RESULTSIn the univariate analysis, onset age (t = 2.292, P = 0.086), PVR (t = 2.181, P = 0.03), size of the prostate in transabdomen ultrasonography (t = -1.987, P = 0.047), experience of the operator (Z = 10.13, P = 0.0015) and the symptom that bladder does not feel completely empty right after urinating (chi(2) = 9.240, P = 0.002) had statistical significance. In the multivariate unconditional Logistic regression analysis, Odds ratio (OR) of the factors were investigated, the symptom that bladder does not feel completely empty right after urinating (OR = 0.557), the score for inflammation (OR = 0.905) and experience of the operator (OR = 0.393) had statistical significance.
CONCLUSIONSThe risk factors for elevating the incidence of post-TURP recurrence are the lower score for inflammation, younger onset age, having the symptom that bladder does not feel completely empty right after urinating, bigger size of prostate in transabdomen ultrasonography, lower post void residual urine volume and poor experience of the operator.
Humans ; Logistic Models ; Male ; Postoperative Period ; Prostatic Hyperplasia ; surgery ; Recurrence ; Reoperation ; Retrospective Studies ; Risk Factors ; Transurethral Resection of Prostate
8.Studied of dry matter accumulation and echinacoside content of Cistanche tubulosa in Huabei plain.
Tai-Xin YANG ; Yue-Xia LU ; Yu-Hai GUO ; Zhi-Xi ZHAI ; Shu-An WANG ; Lai-Qing LU ; Guo-Jian YU
China Journal of Chinese Materia Medica 2006;31(16):1317-1320
OBJECTIVETo give some theory support of Cistanche tubulosa cultivation by searching dry matter accumulation and echinacoside content of C. tubulosa.
METHODDry matter accumulation content of C. tubulosa culturing in Huabei plain was analysed in different growth season of C. tubulosa. Echinacoside content was determined by HPLC.
RESULTDry matter accumulation of C. tubulosa showed "S" variation. Dry matter accumulation increased fastest in September among growing seasons. Dry matter amount was 138.58 g after C. tubulosa grew a year. Dry matter amount decreased significantly along with inoculation time retarded. Echinacoside content was 30.59% when C. tubulosa grew in 5 months, decreased guadully after that, and 9.76% in annual.
CONCLUSIONVariation rule of dry matter accumulation and echinacoside content was found in C. tubulosa that grew one year in Huabei plain.
Biomass ; China ; Cistanche ; chemistry ; growth & development ; Glycosides ; metabolism ; Plants, Medicinal ; chemistry ; growth & development ; Seasons
9.The SARS-CoV 3a and 7a Protein May Enhance the Induction of IFN-?
Chun-E XU ; Ling FU ; Lihua HOU ; ShaoJie WENG ; DaZhi LAI ; JianMin LI ; Ting YU ; ChangMing YU ; Wei CHEN
China Biotechnology 2006;0(12):-
3a and 7a are nonstructural proteins of SARSCoV, which are encoded separately by ORF 3a and ORF 7a in SARSCoV genome. The expression of 3a has been founded in cells infected by virus in vivo or in vitro. Firstly, the pGL3Control vector was reconstructed , the pGL3Enhancer vector deletious of SV40 promoter gene was obtained . Then the IFN? promoter gene was cloned into the pGL3Enhancer vector and pGLIP21, the Luciferase reporter plasmid with IFN? promoter was established. The availability of pGLIP21 was verified by NDV ,the inductor of IFN?, the Luciferase activity was assayed in cells transfected with pGLIP21 by Luminometer. In order to see the function of 3a and 7a protein of SARSCoV,CHO cells expressing 3a or 7a protein were transfected with pGLIP21, the intensity of luciferase activity was analyzed . By analysis, in vitro, 3a and 7a protein of SARSCoV had the similar ability in triggering the expression of Luceferase gene, i.e 3a and 7a protein of SARSCoV could effectively activate the promoter fragment of IFN? gene. This result will help studying the function of 3a and 7a protein and provide a method to study the nosogenesis mechanism of SARSCoV.
10.Surgical therapy of midline skull defect accompanied with frontal sinus injury.
Bing YU ; Feng ZUO ; Jian-Feng LIU ; Lai GUI
Chinese Journal of Plastic Surgery 2012;28(2):105-109
OBJECTIVETo investigate the surgical therapy of midline skull defect accompanied with frontal sinus injury.
METHODS11 cases with midline skull defect accompanied with frontal sinus injury were treated. Free temporal fascia was transplanted to close the top of frontal sinus after curettage of the frontal sinus wall. Then titanium prostheses were used to repair the skull defects at the same stage in 10 patients. 1 patient received skull defect repair at the second stage operation.
RESULTSGood results were achieved in 10 cases. The titanium prosthesis had to be taken out in one case due to frontal sinusitis and the anastomosis of frontal sinus and nasal cavity was performed.
CONCLUSIONSIn patients with midline skull defect accompanied with frontal sinus injury, free temporal fascia could be used to close the top of frontal sinus after curettage of frontal sinus wall. If there is no infection or mild infection in frontal sinus, the skull defect repair could be performed in the same stage. If there is severe frontal sinusitis, the defect repair should be done at the second stage.
Facial Injuries ; surgery ; Frontal Sinus ; injuries ; Humans ; Prosthesis Implantation ; methods ; Skull ; injuries ; Skull Fractures ; surgery ; Titanium