6.The study of biological characteristics of myoblasts co-cultured with Schwann cells
Hua LIAO ; Lei YU ; Shao-Hu XIONG ; Al ET
Chinese Journal of Orthopaedics 2000;0(11):-
Objective In order to understand the relationship between myoblast and Schwann cell,our purpose was to investigate the effects of biological characters of myoblasts co-cul tured with Schwann cells and pro vide the basic theory for constructing artificial muscle involving artificial nerve.Methods After sterilizing with iodine tincture and alcohol,the brachial plexus,sciatic nerve and triceps surae muscle of neonatal SD rat were harvested and peeled off their membranes,blood vessels and fat tissues under operating microscope thor oughly.The nerve and muscle tissues were cut in pieces by microscis sors,and then digested and isolated by collagenase and pancreatin in20and15minutes respectively.DMEM medium was employed to culture my oblasts and Schwann cells.After co-culturing myoblasts of rats with Schwann cells in vitro,the morphological characteristics and the growth condition of two cells were observed under inverted phase contrast mi croscope,the effects of Schwann cells secretion for proliferation of myoblasts were detected by 3 HTdR iso topic tracing and expressed by disintegration per minute(DPM),formation rate of myotubes was counted under micro scope and statistic data was analyzed,the functional differentiation degree of my oblasts affected by Schwann cells was analysed by?-sarcomeric actin immunohistochemistry(SABC)and imaging analysis tech nique.Results Co-cultured myoblasts proliferated,and myotubes ap peared earlier.Comparing with sole-cultured my oblasts,the shape of myobutes from co-cultured myoblasts tended to be elongating and robust.The value of DPM far-exceeded the control group,and reached its peak of 2500(just800for con-trol group).The positive cells of ?-sarcomeric actin appeared in brown red color.However,syn thesis and excre tion of a-sarcome ric actin in co-cultured myoblasts were much greater than control group,and the gray ash value between two groups was of a significant difference.Conclusion Primary rat myoblasts co-cultured with Schwann cells in vitro is beneficial in regulating its the growth,proliferation and the differentiation.
7.Correlation between invasion and metastasis of gastric cancer and serum angiopoietin-2
Shao-ping LIU ; Chun-hua FANG ; Li-qun YU ; Xian-feng ZHANG ; Ya-hua HU
Chinese Journal of Postgraduates of Medicine 2011;34(28):31-34
Objective To study the clinical significance of the serum angiopoietin-2(Ang-2) in the diagnosis,recurrence,invasion and metastasis of gastric cancer.Methods The serum Ang-2 and carcino-embryonic antigen (CEA) levels in 158 patients with gastric cancer (gastric cancer group) and 30 normal controls(control group) were measured by enzyme linked immunosorbent assay(ELISA) technique respectively.The serum Ang-2 and CEA levels were also measured 2 weeks after operation in gastric cancer group and reexamined in the recurred gastric cancer patients in 2 years after operation (recurred and metastasis group).The correlation between the serum Ang-2 level and pathologic c haracterization of gastric cancer was evaluated.Results The serum Ang-2 and CEA levels in gastric cancer group [ (331.8 ±64.3),(42.6 ±37.3)μg/L] and recurred and metastasis group [(318.7 ±72.9),(40.5 ±36.7)μg/L] were significantly higher than those in control group [ (187.4 ± 32.7),(4.2 ± 3.1 )μ g/L] (P < 0.01 ),and the serum Ang-2 level 2 weeks after operation [ (211.6 ± 75.1 ) μ g/L ] was significantly decreased to the control group (P > 0.05 ),while the serum CEA level [ (33.4 ± 30.6) μ g/L ] was still significantly higher than the control group (P < 0.01 ).The sensitivity of the serum Ang-2 for diagnosis of gastric cancer was markedly higher than that of the serum CEA (P < 0.01 ).There was correlation between serum Ang-2 and degree of tumor differentiation,TNM pathological staging,lymphatic metastasis,invasion depth and tumor size (p <0.01 ),but there was no correlation between serum Ang-2 and tissue classification and location of gastric cancer (P> 0.05).Conclusion The serum Ang-2 level is suggested to be a valuable gastric cancer marker and conduce to the diagnosis of gastric cancer,the monitoring of recurrence after operation and evaluation of prognosis.
8.AO elastic intramedullary nailing for the treatment of clavicle fractures.
Hong-wei XU ; Jun-yu HU ; Shao-hua JIA ; Zhong-wei ZHANG ; Sui-liang GONG
China Journal of Orthopaedics and Traumatology 2015;28(2):106-111
OBJECTIVETo evaluate the outcome of Titanium elastic intramedullary nailing (TEN) for the treatment of displaced midclavicular fractures.
METHODSBetween February 2010 and February 2013, 62 patients with displaced midclavicular fractures were treated by TEN, including 27 males and 35 females with an average age of 37.6 years old ranging from 15 to 67 years. The course of disease was from 1 to 9 days (means 2.7 days). Thirty-nine cases were treated by closed reduction and 23 cases by assistant small incision. Based on OTA (the Orthopaedic Trauma Association classification) classification, 31 cases of simple fractures involved 5 cases of 06-A1,15 cases of 06-A2,11 cases of 06-A3; 31 cases of wedge fractures involved 4 cases of 06-B1,12 cases of 06-B2,15 cases of 06-B3. Postoperative pain relief were evaluated by VAS score,and operation time,fracture healing time were recorded. After 6 weeks and after removing internal fixation shoulder joint function was evaluated by Constant score, and shoulder joint function were assessed by Herscovici score after 6 weeks.
RESULTSExcept 2 cases were lost to follow-up, 2 cases did not remove internal fixation, 3 cases' fractures were nonunion, the remaining 55 patients received follow-up for an average time of 11.4 months. The average preoperative VAS score was 5.20±0.71, and it turned to be 1.550.59 at 3 days after operation. The average operative duration was 40 min (15 to 65 min). The average bone healing time was (2.71±0.54) months (2 to 5 months). No difference about bone healing time was found between simple fractures and wedge fractures,and between the patients treated by closed reduction and assistant small incision. According to Herscovici standard, the shoulder function was excellent in 49 cases, good in 4, fair in 1, and poor in 1. Simple fractures achieved better Constant score than wedge fractures at 6 weeks postoperatively, whereas no difference was found at 4 weeks after fixation removal.
CONCLUSIONTitanium elastic nails (TEN) for treatment of displaced midclavicular fractures has advantages of good clinical effect,rapid pain relief,fast recovery of shoulder joint function. For OTA classification type B patients with comminuted fracture can replace steel to achieve very good effect, but to grasp the appropriate.
Adolescent ; Adult ; Clavicle ; injuries ; surgery ; Female ; Fracture Fixation, Intramedullary ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged
9.Establishment and optimization of real-time PCR with SYBR Green Ⅰ for quantification of survivin
Li-Hua HU ; Zheng-Jiang CHENG ; Yi-Rong LI ; Shao-Jun HUANG ;
Chinese Journal of Laboratory Medicine 2003;0(12):-
Objective To establish real-time PCR with SYBR Green Ⅰ for quantification the gene of survivin.Methods The components and conditions of PCR system were optimally determined by fluorescence intensity,cycle threshold(Ct),melting curve,coefficiency and slope of the standard curve.The means to eliminate the contaminating fluorescence of primer-dimers and the mode for Ct value determination were also optimized.Using the developed PCR system,we quantificated the survivin gene in 43 patients with gastric carcinoma.Results The optimized condition for PCR amplification of survivin were 2 mmol/L of MgCl_2,2.5 U/100 ?l of Taq DNA polymerase,0.2 ?mol/L of primers,and the optimized annealing temperatures for PCR were 58℃.The influence of primer dimmer can be eliminated by setting the fluorescence collecting temperature below the Tm of the specific amplicon by 2℃.The second derivative maximum mode,instead of fit point mode,was a feasible method to determine the Ct value for quantification.The sensitivity of this method was 10 copies/?l,and a good linearity was found from 10~1 to 10~4 copies/?l(r = 0.999 7).The inter-experimental coefficient of variation was 1.13%-1.91%,whereas the coefficient of variation between runs was 3.31%-4.50%.Using the optimized PCR system,we quantificated the gene of survivin,the result indicated that survivin gene was amplified in 13.9% of gastric carcinomas.Conclusions The optimal real-time PCR with SYBR Green Ⅰ,as a cost-effective and feasible DNA quantitative method,is fit for quantification of the survivin with satisfactory repeatability and high sensitivity.