Objective To explore the inductive effect of striatal tissue on mouse embryonic stem cells and further analyse the cell source and inductive pattern of this inductive effect. Methods We employed striatal extracts、conditioned medium of striatal astrocytes and conditioned medium of striatal neurons to induce embryonic stem cell to differentiate directly. The differentiated cells were evaluated by morphological observation and TH immunocytochemical staining method. We also performed a quantitative analysis of the results. Results Striatal extracts and conditioned medium of striatal astrocytes had obvious inductive effect on embryonic stem cell.Percentages of three groups were 15%,15.2% and 3% respectively. Conclusions The astrocytes in striatum might have an inductive effect on dopaminergic neuronal differentiation of embryonic stem cells.The determination of inductive factor will be our next research aim.;

Objective To explore the optimal condition of direct differentiation into dopaminergic neurons of embryonic stem cells in serum-free and feeder layer cell free medium. Methods We used the method of phase induction to culture embryonic stem cells. At first, embryonic stem cells were cultured in the serum-free medium with bFGF and LIF so as to realize the direct differentiation from embryonic stem cells to neural precursors. Differentiated cells were determined by nestin immunocytochemical staining. On this basis, we transferred embryonic stem cells to the B27 serum-free medium with IL-1 so as to realize the direct differentiation from neural precursors to dopaminergic neurons. Differentiated cells were determined by TH immunocytochemical staining. Results Approximately 85 percent of cell masses were nestin immuno-positive. The differentiation ratio of dopaminergic neurons was 13%, which increased significantly in comparison with natural differentiation ratio of dopaminergic neurons.Conclusion Without serum and feeder layer cell, we can induce embryonic stem cells to differentiate into dopaminergic neurons effectively by adding different growth factors at different phases, which makes the inductive processes more easily.

Objective To carry out construction of chimeras from embryonic stem cells of outbred KM mice. Methods We isolated embryonic stem cells from inner cell mass of KM mice blastocysts. Then we transferred embryonic stem cells into blastocoele of C57BL/6J inbred mice by microinjection in order to construct chimeric mice. Results The cells isolated from inner cell mass have typical characteristics,i e positive alkaline phophatase staining,normal karyotype,forming embryoid body. Now,we have constructed one chimeric mice successfully. Conclusion Embryonic stem cells isolated from inner cell mass can be used for the chimeras production successfully,which forms the substantial base of transgenic animal model by the way of using embryonic stem cells.

Objective To explore the survival,migration and differentiation of embryonic stem cells after transplantation into striatum and provide advantageous data for feasibility and safety of therapeutic transplantation. Methods We transplanted embryonic stem cells(undifferentiated ESCs and ESCs that had already developed into the stage of neural progenitor cells respectively) into striatum of the rat.Then differentiated cells were determined by morphological observation,Nissl's staining,TH and BrdU immunocytochemistry. Results We found that after transplanted 4 weeks,partially differentiated ESCs could survive and migrate into the surrounding host tissue.Some of them differentiated into TH-positive cells which had Nissl's bodies in cytoplasm.Whereas undifferentiated ESCs couldn't differentiate into TH-positive cells effectively and have the tendency to form tumor.Conclusion When conducting transplantation experiments of ESCs,it's better for ESCs to be induced into the stage of neural progenitor cells first and then transplanted.;

Objective To summarize our experience in the prevention and treatment of accessory hepatic duct injury during operation on biliary tract.Methods The clinical data of 26 cases with accessory hepatic duct were retrospectively reviewed.Results Of 26 cases,the accessory hepatic duct were type I in 38.5%(10/26),and no complications including bile leakage,biliary infection and obstructive jaundice developed after division and ligation of the accessory hepatic duct;26.9%(7/26) were type II,among which,the accessory hepatic duct were injured in 3 cases,but no case developecl complications after relevant treatment;23.0%(6/26) were type III,among which,injury of accessory bile duct occurred in 2 cases.Of them,1 case developed bile leakage and was cured by reoperation.7.7%(2/26) were type IV and 3.9%(1/26) was type V.The cases of type IV and V were not damaged.Conclusions To prevent injury of accessory hepatic duct,pre-and intra-operation identification of the condition is very important,and especially by intraoperative cholangiography.Different types of accessory hepatic duct injury should be treated by different approaches. Accessory hepatic duct of type I might be cut and ligated.Type II accessory bile duct which(enters) the cystic duct and should be protected,but,if damaged,different methods of treatment are used,(depending) on the caliber of accessory hepatic duct.Type III and IV also should be protected,but,when damaged,the accessory hepatic duct should be repaired or performed an internal draining.

Objective To investigate the effect of granulocyte colony-stimulating factor(G-CSF)and its effect on the cognation in the PDGF hAPPV717I transgenic mice of Alzheimer's disease model.Methods Totally 36 PDGF hAPPV717I transgenic mice were randomly divided into two groups:G-CSF group and control group.The G CSF group was subcutaneously injected with 50 μg · kg-1 · d-1 of G-CSF for 7 days.The control group was injected subcutaneously with an equal volume of PBS in parallel.The animals were tested in Morris water maze on the 7th,14th,and 28th days after the last day of the injection,and the escape latency was recorded.Once the test was completed,the peripheral blood was taken to evaluate the effect of G-CSF to induce hematopoietic stem cells (HSCs) via flow cytometry.Then the mice were killed,their brains were quickly removed and frozen on dry ice.With the immunohistochemical staining and double immunofluorescence staining,the neurogenesis could be observed in the model mice.Results We found that G-CSF significantly shortened the escape latencies in PDGF-hAPPV717I transgenic mice compared to controls on the 7th,14th,and 28th day after G-CSF treatment [7 d:(27.19±4.07) s and (46.07±7.21) s,P＜0.000; 14 d:(26.48±5.29) sand (42.99±11.70) s,P＜0.010; 28 d:(24.97±3.61) s and (45.54±9.55) s,P＜0.002)].At the same time,we found that the percentage of CD34+/CD45+ cells in the peripheral blood was 0.358±0.161,0.223±0.038,0.168±0.049 on the 7th,14th,and 28th day after G-CSF treatment,respectively.Compared with the control group (0.073±0.026,0.067±0.034,0.072± 0.037),the percentage of CD34′ /CD45+ cells in the peripheral blood were increased (P＜0.001).BrdU+ cells were found in dentate gyrus (DG) of hippocampus and the cortex of the G-CSF group,where the BrdU+ /Ncstin- and BrdU-/MAP-2+ cells were also detected positively.Conclusions Subcutaneous administration of G- CSF may improve the cognition in APP transgenic mouse model of AD.G-CSF may mediate the proliferation,differentiation of hcmatopoietic stem cells (HSCs).and may induce the neural stem cells into the brain.

Objective To evaluate the diagnostic value of US, ECT, CT and MRI in primary hyperparathyroidism (PHPT). Methods Data of 34 PHPT patients with diagnosis confirmed by postoperative pathology were retrospectively reviewed from January 1, 1990, through March 31, 2004. Results The preoperative diagnosis in 22 out of 25 cases (88%) undergoing preoperative ultrasonography of the neck with a positive result was verified by intraoperative findings. For 99m Tc-MIBI, CT and MRI, the positive results were 95%, 69% and 100% respectively. Conclusion Ultrasonography and 99m Tc-MIBI should be considered as the first choice for preoperative loci localization in patients with PHPT. Ultrasonography and 99m Tc-MIBI in combination is more sensitive and accurate for the localization of PHPT.

Objective To explore the diagnosis and treatment of primary hyperparathyroidism (PHPT). Methods Clinical data of 35 cases of PHPT were retrospectively analyzed. Results 23 out of 26 cases (88.5%) undergoing preoperative ultrasonography with a positive result were verified by intraoperative (findings). For ECT, the positive rate was 21 out of 23 cases (95.5%). Unilateral neck explorations (UNE) was performed in 27 cases of parathyroidoma. Two cases of parathyroid hyperplasia were treated by UNE and the other two cases by bilateral neck exploration (BNE). The procedure for 3 cases of parathyroid carcinoma was the same as that for papillary thyroid carcinoma. Unilateral resection of thyroid and parathyroid was done in a case of parathyroidoma with malignant change. Emergency excision of parathyroidoma, after (emergency) medical management, was performed in a patient with parathyroid storm, and the symptoms (subsided) postoperatively. All cases developed hypocalcemia in various degrees after surgery, but the symptoms were relieved with the use of calcium gluconate. Conclusions PHPT could be diagnosed according to (co-elevated) calcemia and PTH. Ultrasonography and ECT should be considered as the methods of first choice for preoperative localization. UNE of parathyroidoma could be feasible if accurate image localizations are (available). Radionuclear guided parathyroid resection could be performed in the patients with ectopic parathyroid disease or lesions without accurate localization. Aggressive surgical exploration after medical control of (symptoms) is the first choice of treatment when parathyroid storm is diagnosed.

OBJECTIVESTo increase the diagnosis and treatment of ectopic ACTH syndrome.

METHODSThe data of 12 ectopic ACTH syndrome patients treated from 1985 to 1999 were retrospectively analyzed.

RESULTSTwelve patients were diagnosed as having ACTH syndrome by endocrinary test and primary tumors were ascertained by imaging examination. Follow-up from 7 months to 8 years showed 3 out of 5 patients with radical resection of primary tumor died. One patient with bilateral adrenorectomy was still alive. All patients received only chemotherapy except one died.

CONCLUSIONSPatients with Cushing's syndrome should be evaluated by endocrine test and followed up by imaging screen examination. The key points to increase treatment effect include early detection, localization and resection of primary tumors.

In order to enhance the efficiency of sheep somatic cell nuclear transfer, we used a chemically assisted enucleation with colchicine to study the effects of the concentration of colchicine, the incubation time of oocytes in colchicine and the maturation time of oocytes on the enucleation rates and the development of reconstructed embryos. The results showed that 1) there were no significant differences in the rates of cytoplast protrusion and enucleation between oocytes that were incubated in colchicine (0.4 microg/mL) for 0.5 h and oocytes that were incubated in colchicine (0.4 microg/mL) for 1 h, and the rate of cytoplast protrusion can be 85.4% while the rate of cytoplast enucleation is 100%. 2) There was no significant difference in oocyte enucleation between oocytes treated with medium containing 0.2 microg/mL colchicine for 0.5 h and oocytes treated with medium containing 0.4 microg/mL colchicine for 0.5 h. 3) A maturation time of 18-23 h did not affect the rates of cytoplast protrusion and enucleation by chemically assisted enucleation, whereas the rate of enucleation of oocytes by blind enucleation was found to decrease with a prolonged incubation time. 4) The development rates of reconstructed embryos could not be influenced by these two enucleation methods, increased from oocytes matured for 21-23 h. These results demonstrate that sheep oocytes can be enucleated fast and effectively by optimized colcholine chemically assisted enucleation, which can enhance the enucleation rate of sheep oocytes and the early development of reconstructed embryos in vitro.
Animals ; Cloning, Organism ; methods ; Colchicine ; pharmacology ; Embryo, Mammalian ; embryology ; Female ; Nuclear Transfer Techniques ; veterinary ; Oocytes ; cytology ; drug effects ; Sheep