273 spine apparatus were observed under electron microscope. The results were as follows: 1.The flat-type sac of 5 types of the sac which constituted the spine apparatus covered the majority. Most of them were arranged parallelly and a few dispersively. The horseshoe or braid shaped arrangement was also seen in some cases. 2.The number of the sac varied from 3 to 6 in most of the cases, and a few over 10. Sometimes 2 or 3 spine apparatus can be seen within a single dendritic spine. 3.The spine apparatus often locates in the dendritic spine. In some cases the spine apparatus can be seen within the dendrites or in the juncture of dendritic trunk and dendritic spine 4.The morphological structure of the dense bands is comparatively complex. Most of the bends are narrow, some are broad, fine granular or mist-like in shape. The dense bends are located in between the adjacent sacs mostly and in few cases on one side of the spine apparatus. In some cases, one end of the dense band is fused with the sac.

Objective: To evaluate the effect of parallel scalp acupuncture therapy on hemodynamic function of cerebral circulation in post-stroke patients. Methods: Twenty post-stroke patients were taken as our objects. The cerebral circulation indices before and after parallel acupuncture therapy, were detected with KF-3000 Brain Circulation Analyze, to compare the change of hemodynamics indexes before and after parallel scalp acupuncture. Results: After the therapy, minimum and mean blood velocity of carotid artery (Vmin and Vmean) increased significantly (P<0.001); peripheral resistance (Rc) decreased significantly (P<0.001). And the other indices had no significant differences. Conclusion: Parallel Scalp Acupuncture can improve the state of cerebrovascular autoregulation, raise the velocity of blood, decrease the peripheral resistance, and increase the steady energy, total energy and the ratio of kinetic energy to total energy, decrease the ratio of oscillatory kinetic energy to total kinetic energy.

Objective To explore the expression characteristics of vascular endothelial growth factor (VEGF) receptor (VEGFR).Methods The 123I-VEGF165 and 123I-VEGF121 were marked to human umbilical vein endothelial cell (HUVEC),several human tumor cell lines (HMC-1,A431,KU812,U937,HEP-1,HEP-G2,HEP-3B and Raji),a variety of human tumors and adjacent non-neoplastic tissues as well as peripheral blood cells.Then,the specific binding site maximal binding capacity (Bmax),dissociation constant (Kd) and concentration of 50％ required specific binding (IC5o) were analyzed.The affinity,quantity and specificity of different cells combined with 123I-VEGF165 and 123I-VEGF121 were judged.Results Two kinds of analogous 123I-VEGF165 binding sites on the surface of HUVEC were found.While,there was only one kind of 123I-VEGF121binding site.123I-VEGF121 was found on the special cell lines (HUVEC,HEP-1 and HMC-1) and special early tumors (early melanoma,ductal breast cancer,ovarian cancer and meningioma).Compared with peripheral blood cells and adjacent non-neoplastic tissues,the number of VEGFR of tumor cells was bigger.Among the 123I-VEGF165 marked VEGFR,the Bmax value of early melanoma,ductal breast cancer,hepatocellular carcinoma,papillary thyroid carcinoma,ovarian carcinoma,renal cell carcinoma were 45 ± 13,13 ± 3,25 ±8,5 ±2,42 ± 12,20 ±6,respectively.While among the 123I-VEGF121 marked VEGFR,the Bmax value of early melanoma,ductal breast cancer,ovarian carcinoma were 30 ± 8,8 ± 3,20 ± 6.123I-VEGF165 and 123I-VEGF121 had specific binding capacity with a variety of human tumor cells and tissues.Compared with 123I-VEGF121,there were more different kinds of tumor cells could be bound to 123I-VEGF165 with higher capacity.Conclusion 123I-VEGF165 may be a potential target of tumor imaging in vivo,and it is expected to be used to diagnose and treat tumors.

The choroid plexus of the cat, rabbit and rat have been investigated by means of scanning elecron microscopy. The surface appearance of choroid plexus of these animals exhibited pleomorphism. The free surface of epithelial cells are covered with dense microvilli, single long cilium, clusters of cilia, bulbous protrusions and flower-like structure. They are scattered throughout the ventricular surface of the choroid plexus. High magnification scanning electron microscopy reveals that the population of microvilli consist of slender finger-like microvilli and microvilli with the bleb-like protrusion. Flower-like structure is the clusters of microvilli. On the surface of the choroid plexus of these animals, the Kolmer cells were observed. According to the number of processes, the Kolmer cells of cat, rabbit and rat may be divided into four main types. The ultra-architectural organization of choroid plexus of the cat, rabbit and rat is quite similar, but the number of bulbous protrusions in the cat and clusters of cilia in the rat occurs more.

Objective To explore the isolation, culture of nestin positive cells of the pancreas in newborn rats. Methods The whole pancreas of neonatal rats were digested with collagenase, followed by incubation under pH 7. 6 serum RPMI 1640 for 24 - 36 h and then under pH 7. 4 serum free RPMI 1640 (bFGF.EGF 20 ng/ml.1% N2) for 18-24 d. The expression of insulin.glucagon and nestin were detected by immunofluorescence technique. Nestin.CK19 were detected by RT-PCR. Resides Cells attached to the surface of plates after 24 h incubation under pH 7. 6 condition, and islet-like masses were obtained after 18 -24 d incubation. A monolayer of cells grew out after 24 h of passage of islet-like masses. Nestin positive cells was detected after 24 - 36 h incubation, with no expression of insulin and glucagon. Positive cells of insulin and glucagon were detected in islet-like masses after 24 h passage. Nestin positive cells were detected by RT-PCR in islet-like masses after 24 h passage, but no CK19. Conclusion Insulin and glucagon were expressed in islet-like masses after passage. Nestin positive cells in the pancreas of neonatal rats possessed character of islet stem cells.

Objective To explore the method of isolation, identification and in vitro differentiation of islet stem cells in neonatal rat. Methods The whole pancreata of neonatal rats were digested with collagenase, then,under the (condition) of pH 7.4～7.6,the digested tissue fragments were cultured with serum and serum-free RPMI 1640 (( bFGF), EGF, N2).The whole formation process of new islet cell-like cell masses was examined.The insulin (release) test was used to detect islet function. The expression of nestin was tested by immunocytochemistry. Results The nestin positive cells can be found within 36 h of culture of the digested pancreatic fragments.After addition of bFGF,ECF,N2,nestin positive cells proliferated fast and formed new islets-like cell masses after 18～24 d, and (insulin)ssion could be confirmed. Conclusions The nestin positive cells of pancreatic cells possess the character of islet stem cells, and can form islet like cell masses through culture in vitro.

To clone and sequence the human tissue kallikrein gene of Chinese, and to construct eukaryotic expression recombinant of KK, total RNA was extracted from human pancreas and human tissue kallikrein gene cDNA was amplified by PCR after reverse-transcription by using Oligo(dT) primer. The original kallikrein cDNA was recovered and filled with Klenow enzyme and inserted into KS plasmid. After restriction endonuclease digestion, KK cDNA was sequenced by ABI377 analyzer. Then the KK gene was amplified from pBluescript KSKK and inserted into pcDNA3. A sequence comparison showed that the cloned kallikrein gene was only one nucleotide different from that reported in the Genbank. The coding amino acid was Asp in the Genbank gene, while the coding amino acid of Chinese kallikrein gene was Asn. The KK cDNA fragment was inserted into the eukaryotic expression vector pcDNA3. The cloned kallikrein gene and the pcDNA3KK can be used for further study in gene therapy.. .

Objective The incidence rate of sacrococcygeal tumor is low , however , due to its special position , clinical symptoms are inclined to be more severe .The aim of the article was to explore the methods and therapeutic effects of operative treat-ment for sacrococcygeal tumors . Methods A retrospective analysis was made on 44 cases of sacrococcygeal tumors treated in our hospital from January 2008 to December2012 .Of all the cases , there were 11 chordomas , 9 neurinoma , 4 malignant fibrous histioto-ma, 4 giant cell tumor, 7 metastases tumors and 9 others.All the patients had definite pathological diagnosis after operation .Operative treatments involved simple resection of rumors , simple resection of lesions and resection of lesions plus screw-rod system internal fixa-tion. Res ults No patients died in perioperative period .Follow-up was made in all patients for the average time of 3.3 years(4 months to 59 months).Except for 4 patients'delayed healing of incision operation and 1 patient′s healing of operation incision after de-bridement , all the other patients healed after enhancing the wound dressing , among which there are 3 cases of chordoma recurrence , 1 case of giant cell tumor recurrence , 3 death cases of malignant fibrous histiotoma in 2 years and 1 case alive with tumor .All patients with metastases tumor died in 3 years.Except 1 patients with osteosarcoma alive with tumor , all patients′symptoms had been alleviated after operation. Conclusion Chordoma and giant cell tumor still have a high recurrence rate after operation . The extent of rumor resection and nerve preservation are determined by the range of tumors.Preoperative embolization of the internal iliac artery can clearly reduce bleeding and improve the operation safety .

To clone and sequence the human tissue kallikrein gene of Chinese, and to construct eukaryotic expression recombinant of KK, total RNA was extracted from human pancreas and human tissue kallikrein gene cDNA was amplified by PCR after reverse-transcription by using Oligo(dT)primer. The original kallikrein cDNA was recovered and filled with Klenow enzyme and inserted into KS plasmid. After restriction endonuclease digestion, KK cDNA was sequenced by ABI 377 analyzer.Then the KKgene was amplified from pBluescript KSKK and inserted into pcDNA3. A sequence comparison showed that the cloned kallikrein gene was only one nucleotide different from that reported in the Genbank. The coding amino acid was Asp in the Genbank gene, while the coding amino acid of Chinese kallikrein gene was Asn. The KK cDNA fragment was inserted into the eukaryotic expression vector pcDNA3. The cloned kallikrein gene and the pcDNA3KK can be used for further study in gene therapy...

Antibody-drug conjugates(ADCs)are commonly heterogeneous and require extensive assessment of exposure-efficacy and exposure-safety relationships in preclinical and clinical studies.In this study,we report the generation of a monoclonal antibody against monomethyl auristatin E(MMAE)and the development,validation,and application of sensitive and high-throughput enzyme-linked immunosor-bent assays(ELISA)to measure the concentrations of MMAE-conjugated ADCs and total antibodies(tAb,antibodies in ADC plus unconjugated antibodies)in cynomolgus monkey sera.These assays were suc-cessfully applied to in vitro plasma stability and pharmacokinetic(PK)studies of SMADC001,an MMAE-conjugated ADC against trophoblast cell surface antigen 2(TROP-2).The plasma stability of SMADC001 was better than that of similar ADCs coupled with PEG4-Val-Cit,Lys(m-dPEG24)-Cit,and Val-Cit linkers.The developed ELISA methods for the calibration standards of ADC and tAb revealed a correlation be-tween serum concentrations and the OD450 values,with R2 at 1.000,and the dynamic range was 0.3-35.0 ng/mL and 0.2-22.0 ng/mL,respectively;the intra-and inter-assay accuracy bias％ranged from-12.2％to-5.2％,precision ranged from-12.4％to-1.4％,and the relative standard deviation(RSD)was less than 6.6％and 8.7％,respectively.The total error was less than 20.4％.The development and validation steps of these two assays met the acceptance criteria for all addressed validation parameters,which suggested that these can be applied to quantify MMAE-conjugated ADCs,as well as in PK studies.Furthermore,these assays can be easily adopted for development of other similar immunoassays.