Method of selecting points in foot-hand same-name meridians was used to treat 24 cases of Osteoarthritis of knee joint, and the therapeutic results showed 15 cases were cured, 4 cases got significant effect and 4 cases got improvement.

Objective To simulate the hemodynamic feature in cerebral bridging veins (BVs), in order to provide a morphologic basis for the pathogenesis explanation and imaging diagnosis of cerebral venous thrombosis (CVT). MethodsTotally 6 human cadavers (12 sides) were examined in this study. Each head of the cadavers was injected with blue-coloured latex via the superior sagittal sinus (SSS) and internal jugular veins. The diamter and the angle of BVs entering SSS were measured. Based on the data of cadavers and computational fluid dynamics software pack, the hemodynamic models were established. The wall shear stress (WSS) was carefully studied and compared between different models. Results The total of 137 BVs formed two clusters along the SSS: anterior group and posterior group. Compared with anterior group BVs, the diameter of posterior group BVs was large, and the angle was smaller. In 137 models,when the diameter of a BV was more than 1.2mm, and the angle was between 65 and 105 degree, the local WSS decreased in the downstream wall of SSS. When the diameter of a BV was more than 1.2mm, and the angle was less than 65 degree, the local WSS decreased in the downstream wall of SSS and the upstream wall of BVs. The minimum WSS in BVs was 63% of the minimum WSS in SSS. Compared with the anterior group BVs, the minimum WSS in the wall of posterior group BVs was samller, and the distance from the minimum WSS to the dural entrance was longer. Conclusion CVT occurs easily when the diamter of a BV is more than 1.2mm and the angle is less than 65 degree. The embolus forms early in the upstream wall of BVs entering the posterior part of SSS.

The flushing pump which is applied to clean operative wound has no temperature controlling function up to now, and doctors have to prepare the flushing fluid that has previously been warmed. The flushing pump system with medical constant temperature designed in our laboratory can absorb flushing fluid at the room temperature, and then eject flushing fluid with the temperature in accordance with the requirements of operations at a controlled constant flow rate. The system combines flow rate control with temperature control functions. The flushing pump system includes flushing part, temperature controlling part, key inputting part, liquid crystal displaying part and exceptional situation monitoring part. The present paper introduces the design method and principle of each part of the system at first, and then gives the debug method of all the system parameters. Finally the paper discusses the performance of the system according to the result of the experiment.
Equipment Design ; Equipment and Supplies ; Temperature

In surgery operations, wound should be cleaned with warm sterilized saline solution. In order to reach rapidly warming the washing solution from the room temperature during the surgery, we designed a thermostatic medical infusion pump. The present paper mainly presents researches on the two temperature control methods in the standby mode and in the flushing mode of the system. In the standby mode, the traditional proportional-integral-derivative (PID) control algorithm was adopted. In the flushing mode, dynamic characteristics of the system was changed in real time, which made the thermostatic control process more complex, and the fitted control function combined with the PID control algorithm was adopted in this mode. The temperature control parameters were adjusted in real time according to the initial temperature and the flow rate of the washing solution to obtain a constant temperature of the washing solution, no matter how the initial temperature and the flow rate are changed. The experiment results showed that this kind of control system performed well with a high accuracy.
Algorithms ; Equipment Design ; Infusion Pumps ; Sodium Chloride ; Solutions ; Temperature

Objective To further improve the morphological materials of AGs by micro-dissection, histology and CT, we observed the arachnoid granulations (AGs) in middle cranial fossa. Methods Thirty-three adult cadaveric heads were used for microsurgical dissection;Histological sections of AG specimens from 3 cadaver heads were examined. Forty patients who had both normal conventional brain CT and computed tomographic venography (CTV) were retrospectively reviewed. Results In middle cranial fossa the AGs occur in the following situations in order of frequency: the middle meningeal sinus, sphenoparietal sinus, lateral foramen rotundum and cavernous sinus. AGs usually show round, oval in shape and irregular in shape. AGs can be divided into individual type and leaflet type under light microscope. The numbers of AGs were observed by microanatomy and CTV were 8.72 and 3.52 respectively. The AGs of cavernous sinus was not localized precisely on CTV. Conclusion Study of the AGs in the middle cranial fossa systematically and comprehensively enriches anatomy and image knowledge. It is helpful in neurosurgical planning and choosing operalion procedure to avoid postoperative complications.

BACKGROUND:Human mesenchymal stem cells (hMSCs) become aging and even die after several passages. Some investigations have shown that telomere has a close correlation with life span of the cells. Whether the ectopic expression of human telomerase reverse transcriptase (hTERT) could induce the activity of the telomerase, maintain the length of telomere, and finally prolong the life cycle of MSCs without losing their multipotent differentiation capacity is still uncertain.OBJECTIVE: To observe the influence of the ectopic expression of hTERT on the telomerase activity and cell life cycles of hMSCs.DESIGN: Repetitive measurement trails.SETTING: Research Center of Stem Cell, Zhengzhou University Medical College.MATERIALS: The experiment was conducted in the Research Center of Stem Cell, Zhengzhou University Medical College from October 2003 to December 2005. hMSCs were obtained from 20 healthy donators from the Department of Pediatric Surgery and Outpatient, the Third and First Affiliated Hospitals of Zhengzhou University. Enhanced green fluorescent protein plasmid (pEGFP-C1) and pEGFP-hTERT were provided by Dr. Chantal Autexier of Canada. DH5α strain provided by Dr. Hou Wei-hong, the Key Molecular Medical Laboratory of Zhengzhou University Medical College.METHODS.: Under sterile condition, 2 mL bone marrow of sternum of healthy donors were harvested, and prepared after centrifugalization,dilution and passage.① Transfection of pEGFP-hTERT into hMSCs and the screening and amplification of resistance cloning:The 5th passage cells were seeded in a 24-well plate,and transfected by pEGFP-hTERT with lipofectamine method.The cells were divided into four groups including untransfected group,lipofectamine group,pEGFP-C1 group and pEGFP-hTERT group. Resistance cloning screen and amplification was performed by G418. ②hTERT mRNA expression and detection of telomerase activity:RT-PCR and PCR-ELISA were used to detect the hTERT mRNA expressions of the fifth passage hMSCs transfected with pEGFP-hTERT, and pEGFP-C1, the untransfected tenth passage hMSCs and K562 cells (positive control), and the telomerase activity of the fifth and thirtieth passage hMSCs transfected with pEGFP-hTERT,the fifth pEGFP-C1-transfected cells and the tenth passage untransfected cells. ③Karyotype analysis of hTERT-transfected MSCs: Chromosome analysis was performed by conventional Giemsa staining.④Inducement of differentiation from telomerase-positive MSCs into neuron-like cells and RE-PCR identification:The transfected MSCs were cultured in a medium containing epidermal growth factor and basic fibroblast growth factor, which could induce the cells differentiate into neuron-like cells. The culture solution was changed every 3 days, and the changes in cell growth condition and morphologic characteristics were observed under an inverted microscope. The microtubule associate protein (MAP2) and neurofliament subunit M (NF-M) were identified by RT-PCR.MAIN OUTCOME MEASURES:①hMSCs transfection with different kathion liposomes and the screening and amplification of resistance cloning; ②hTERT mRNA expressions of each group and detection of telomerase activity; ③Karyotype analysis of pEGFP-hTERT-transfected MSCs; ④ Induction of differentiation from telomerase-positive MSCs into neuron-like cells and RE-PCR identification.RESULTS: ①With the decrease of G418 concentration, the cells in the untransfected and lipofectamine groups died, and stably EGFP expressed MSCs were obtained; after G418 screening, there was a cell clone undergone 35 passages and continued to proliferate, whose appearance and growth characteristics were similar to the untransfected MSCs observed under inverted microscope. ②The fifth passage pEGFP-C1-transfected hMSCs and tenth passage untransfected hMSCs remained telomerase-negative, but the K562 and fifth passage hTERT-transfected cells showed positive telomerase activity. ③The telomerase activity of the fifth and thirtieth passage hTERT-transfected cells was positive. ④The hTERT-MSCs at passage 10, 20 and 30 had 23 pairs of chromosomes, and two X chromosomes. So they were still normal diploid with normal chromosome appearance and number. ⑤Many hTERT-transfected MSCs had the typical appearance of neuron-like cells. RT-PCR analysis showed that th expressions of MAP2 and NF-M were increased.CONCLUSION:Ectopic expression of the hTERT gene is found in hMSCs,and can induce the telomerase activity of hMSCs.The ectopic expression of the hTERT gene in hMSCs could extend the life spans of cells and maintain their multipotent differentiation capacity.

BACKGROUND:Currently,there is not a standard method for in vitro culture and large scale amplification of umbilical cord blood mesenchymal stem cells(UCB-MSCs).OBJECTIVE:To investigate the isolation,purification and culture of UCB-MSCs in vitro,and to detect its surface marker variation.METHODS:The monocytes were harvested from UCB using 1.077 g/cm3 lymphocytes separating solution and density gradient centrifugation,followed by incubation in an incubator containing 5%CO2 at 37℃.The cell morphological changes were observed at different time points and the expression of surface marker was detected using flow cytometry.RESULTS AND CONCLUSION:The monocytes isolated from the UCB grew initially into numerous hematopoietic cell clones,most of which were granulocyte/macrophage colony-forming units and burst forming unit-erithroid,increasing by(37.1±2.3)and (10.4±1.7),respectively.Switzerland staining showed most of them were granulocyte clones(80,1±85.2)%,next was erythroid clones(14.2±1.8)%.At 7 days after culture,some shuttle fibroblast-like cells and fiat osteogenic-like cell spread the whole plastic well.At 14 days after culture,flow cytometry showed CD38+ cells accounted for 1.64%,and CD34+/CD38+ cells accounted for 1,71%,and CD34+/CD38- were 0.55%.PI+ and Annexin-V+ cells accounted for 0.05% and 0.18% respectively.At 21 days after culture,CD38+,CD34+/CD38+ and CD34+/CD38- cells were 74.32%,1.61%,and 0.24%.The results reveled that UCB-MSCs can be isolated and cultured in vitro.

AIM: To observe the inhibitory effects of polypeptide from Buthus martensii venom (PBMV) on human tumor cell lines and the influence of PBMV on cell cycle migration, expression of tumor-suppressor gene p53 and the membrane potential of CNE-2Z cells. METHODS: MTT colorimetric method, the colony formation and mitosis index, flow cytometry assay and intracellular recording with glass microelectrodes were used. RESULTS: PBMV had obvious cytotoxicity on several tumor cell lines. The cells grow was inhibited by PBMV, colony formation rate and mitotic index of tumor cells were reduced and the number of polykaryocyte was decreased. CNE-2Z cells in S phase were reduced evidently after they were treated with PBMV (P

AIM: To construct a recombinant retrovirus vector carrying hTERT for establishing UCBMSCs with hTERT(hTERT-MSCs) to overcome their limited life span and detecting whether telomerized UCBMSCs line maintained long-term self-renewal and differentiation capacity.METHODS: The whole cDNA was generated by PCR amplifications from the plasmid pEGFP-hTERT-C1.The hTERT segments were subcloned into pLNCX2.The target cells were infected with these retroviral particles.The stably transfected cells were selected by neomycin and expanded life span which were designated hTERT-MSCs was observed.The expression of hTERT in mRNA level was detected by RT-PCR and the telomerase activity was measured by TRAP(PCR)-ELISA assay.The hTERT-MSCs were induced with 5-azacytidine to cardiac muscle cells and the specific marker of myocardiocyte was detected.RESULTS: The constructed plasmids were digested with restriction endonucleases(BglⅡand NotⅠ).Two characteristic segments including 6.1 kb and 3.6 kb were obtained.The hTERT-MSCs expressed hTERT in mRNA level.The telomerase activity of hTERT-MSCs was positive.The growth kinetics of hTERT-MSCs was higher than those in UCBMSCs.The hTERT-MSCs were induced to myocardiocyte.CONCLUSION: The hTERT recombinant retrovirus vector has been successfully constructed.The hTERT gene activates the telomerase and prolongs the life-span of cells.No effect of hTERT gene on some type of differentiation potential of MSCs is present.

The experiment was performed at Basic Medical College and First Affiliated Hospital, Zhengzhou University from September 2004 to December 2005. Totally 60 Kunming mice were divided into 5 groups randomly: ①blank control group (n =15) and simple radiation group (n =15). The mice were given 0.2 mL sterile saline by intraperitoneal injection. ②antineoplastic polypeptide from Buthus Martensii Venom (APBMV) group (n =10) and APBMV plus radiation group (n =10) received 0.2 mL APBMV according to prepared concentration by intraperitoneal injection. ③Katsutoxin extract Ⅲ plus radiation group (n =10) received 0.2 mL katsutoxin extract Ⅲ by intraperitoneal injection every other 5.5 hours for 7 days. After 24 hours from the last injection, the mice were endured 60Co g ray radiation (80 cm, 7.5 Gy irradiation dose, 0.27 Gy/min dose rate). Then katsutoxin extract Ⅲ was given same as above for 7 days. Then bone marrow was extracted to be cultured to colony-forming unit-granulocyte and monocyte (CFU-GM). The findings showed that colony amount of APBMV plus radiation group and katsutoxin extract Ⅲ plus radiation group was obviously more than that of simple radiation group [(32?5),(27?3),(2?1)pieces/well,P