1.Gene expression of growth associated protein 43 in activated Schwann cells
Jie LAO ; Liangfu JIANG ; Yudong GU
Chinese Journal of Orthopaedics 2001;0(06):-
Objective To compare activated and normal Schwann cells in GAP43 gene expression. Methods 10 male SD rats, weighed from 100 g to 120 g. The right median nerve of SD rats was transected at the axillary level and was buried in muscle for predegeneration. 1 week later, the distal segment of the transected right median nerve with 1 cm long was harvested. The untreated left median nerve was harvested as control with same length. The epineurium of nerve was stripped, then the nerve tract was cut to small pieces. Schwann cells were obtained by way of double kinases digestion with 0.25% trypsin and 0.03% collegenase. The right median nerve was activated with additional liquid during digestion so as to obtain the activated Schwann cells. The normal Schwann cells were harvested from left median nerve. rt-PCR was used for GAP43 gene enlargement. mRNA was distilled from activated Schwann cells and untreated Schwann cells respectively. Then the mRNA was reversely transcripted to cDNA with SuperScriptTM, and cDNA worked as template for PCR enlargement. The product of PCR was separated with 1% agarose gel electrophoresis for 40 -50 min and stained with SYBR Green Ⅰnucleic acid gel. Fluorescence intensity of GAP43 PCR products was measured and then compared between the experiment group and control group. Results The Fluorescence intensity of GAP43 PCR product of activated Schwann cells was higher than that of normal Schwann cell. There was significant difference (P=0.003, Paired t test). It indicated that GAP43 mRNA of activated Schwann cells was much more than that of the normal Schwann cells. Conclusion GAP43 gene expression is up regulated in activated Schwann cells in contrast to normal Schwann cells. Activated Schwann cells secreting more GAP43, which may be one of the important mechanisms in promoting nerve regeneration.
2.Growth rule of activated Schwann cells cultured on Chitosan-collagen film
Jiyin HE ; Jie LAO ; Yudong GU ; Liangfu JIANG ; Jifeng LI
Chinese Journal of Tissue Engineering Research 2006;10(33):-
BACKGROUND: New-type tissue engineering materials and post-proliferation Schwann cells are implanted into biosynthesis tube for repairing peripheral nerve defect, which are two great developments in the field of artificial biomaterial tube.OBJECTIVE: Taking chitosan-collagen as scaffold, activated Schwann cells as seed cells, we are in attempt to observe the affinity between them as well as growth rule of activated Schwann cells on Chitosan-collagen, so as to provide basis for pre-construction of artificial nerve.DESIGN: Open experiment.SETTING: Department of Hand Surgery, Huashan Hospital Affiliated to Fudan University.MATERIALS: This experiment was conducted at the Key Laboratory of Hand Function Reconstruction, Ministry of Public Health from July 2003 to December 2003. Four male SD rats, of clean degree, were used in this experiment. Chitosan-collagen film was made in Qisheng Biomaterial Technique Institute, Shanghai, Schwann cells activator solution was made in our laboratory (self-made).METHODS: After rats were anaesthenia, the sciatic nerve was cut off to perform predegeneration for 7 days. Another anaesthenia later, the rats were euthanized. Both sides of sorciatic nerves were cut off quickly and put in the D-HANK's solution containing penicillin and streptomycin.Epineurium was eliminated and chipped into 1 mm pieces, then put in the centrifuge tube containing 5 g/L trypsinase and 0.6 g/L collagenase. 0.5 mL activator solution every 2 mL liquid was added and the activated Schwann cells were harvested with the way of two-step enzymolysis. 2×107 L-1 activated Schwann cells in 200 μL were inoculated to Chitosan-collagen film and Petri dish . Two weeks later, cellular growth was observed under phase contrast microscope and scanning electron microscope. Cellular purity was identified with S-100 staining.MAIN OUTCOME MEASURES: ① Drawing cell growth curve and confirming in vitro doubling time. ②Observation of activated Schwanri cells under an inverted phase contrast microscope. ③ Observation of activated Schwann cells inoculated on Chitosan-collagen film under scanning electron microscope.RESULTS: ① Confirmation of in vitro doubling time: Concentration of activated Schwann cells inoculated on both Chitosan-collagen and Petri dish was 2×107 L-1, the final concentration was up to 3.0×108 L-1 and 2.0×108 L-1 respectively 2 weeks later. Doubling time of activated Schwann cells cultured on Chitosan-collagen film was 4 days calculated according to DT=(t-t0) lg2/(lgn-lgn0). ②Observation of activated Schwanncells under an inverted microscope: 24 hours later, the activated Schwann cells inoculated to Petri dish mostly changed from spherical to long shuttle-shape,mutation appeared and most were two-pole shape, fewer were three-pole shape; Morphologically, there was no significant difference between activated Schwann cells inoculated on Chitosan-collagen film and on Petri dish. Activated Schwann cells inoculated to Chitosan-collagen film were like "words cayed on the sand" under phase contrast microscope and the purity was over 95%. ③ Observation of activated Schwann cells inoculated to Chitosan-collagen under scanning electron microscope: Most of activated Schwann cells grew in the introcession of Chitosan-collagen or closely to surface of Chitosan-collagen, presenting regular head-to-end connection and adhesion to Chitosan-collagen film. The cell body was fusiform,with diameter of 4-6 μm, 60-80 μm in length. Cells were shuttle-shape with some small branches. Morphology of Chitosan-collagen film was still complete at week 1.CONCLUSION: There exists great affinity between Chitosan-collagen film and high-purity activated Schwann cell; so tissue-engineering scaffold made of the two components probably promote peripheral nerve regeneration.
3.121 drug-resistance analysis and prevention of MRSA in nenrosurgery intensive care unit
Yong LIU ; Haijun YAO ; Jin HU ; Yanqun JIANG ; Liangfu ZHOU
Chinese Journal of Primary Medicine and Pharmacy 2008;15(4):-
Objective To understand the drug-resistance of MRSA patients in neurosurgery intensive care unit,raise the prevention of MRSA and provide doctors the basis for controlling it. Methods The 5 year(20012005) MRSA patients were tested by Kirby-Bauer in neurosurgery intensive care unit of a third-tier general hospital in Shanghai. Statistic and analysis the drug-resistance of the patients. Results The rates of 121 MRSA drug-resistance to penicillin G,erythromycin, ciprofloxacin, amikacin and the cephalosporins are 92.3 % to 100 %, totally senaitire to teicoplanin and vancomycin and lower drug-resistance to rifampin,netilmicin and fosfomycin, but it rapidly raised from 10.0 % (2001 ) to 95.2 % (2005) to sulfamethoxazole. Conclusion It is time to take care of the drug-resistance of MRSA. Prevention and use antibiotics properly are the important ways to decrease the hospital infection and to improve the quality of recovered.
4.Comparative Study on Acupoint Application Versus Acupoint Injection for Chronic Bronchitis
Feng WANG ; Luomin XIA ; Chen QIAN ; Xinping JIANG ; Liangfu QIN
Shanghai Journal of Acupuncture and Moxibustion 2016;35(5):530-533
Objective To observe the clinical efficacies of acupoint injection versus acupoint application in treating patients with chronic bronchitis due to lung-qi deficiency in remission stage.Method Totally 163 patients with chronic bronchitis were randomized into group A of 78 cases and group B of 85 cases. Group A was intervened by acupoint application, while the control group was by acupoint injection. The syndrome score of traditional Chinese medicine (TCM) was evaluated before and after treatment, the attack frequency of cold, quality of life, and pulmonary function were observed in the 6-month follow-up.Result The TCM syndrome scores were significantly changed after treatment in both groups (P<0.05). There was no significant inter-group difference in comparing the attack frequency of cold in the 6-month follow-up (P>0.05). The scores of quality of life (FS1, FS2, FS3, FS4 and TMS) in group A were significantly changed after treatment (P<0.05). The FS1 score in group B was remarkably changed after treatment (P<0.05). The parameters of pulmonary function (FEV1.0%, V50, V25, FVC, FEV1.0) were significantly changed after treatment in both groups (P<0.05).Conclusion Acupoint application and acupoint injection both can significantly improve the symptoms, quality of life, and pulmonary function in chronic bronchitis.
5.The outcome of anterior transposition of the ulnar nerve in treatment of cubital tunnel syndrome with endoscopically assisted
Yipeng ZHANG ; Weiyang GAO ; Anyuan WANG ; Xueguan XIE ; Liangfu JIANG ; Feiya ZHOU ; Jian DING
Chinese Journal of Microsurgery 2012;35(3):204-206,后插10
Objective To investigate the outcome between endoscopically assisted and routine anterior transposition of the ulnar nerve for treatment of cubital tunnel syndrome.Methods From Februray 2008 to June 2010, forty-four patients with cubital tunnel syndrome were treated with routine anterior subcutaneous transposition (routine group,28 cases) and endoscopically assisted anterior subcutaneous transposition (endoscope group,16 cases).The operate time,drug administration,scar and postoperative hospital stay were compared.The patients were followed 1-12 month postoperatively,postoperative time back to work and function of ulner nerve were recorded.Results The results of endoscope group were as follows: operative time was (67.20 ± 19.69)min; postoperative scar length was (1.5% ± 0.58) cm; rate of administration of anodyne was 6.3%; postoperative hospital stay was (2.4% ± 1.42) days; postoperative time back to work,(14.6 ± 4.69)days; the results of open surgery group were as follows:operative time (62.8% ± 11.06) min; postoperative scar length was (8.7% ± 1.42) cm; rate of administration of anodyne was 42.8%; postoperative hospital stay was (5.7% ± 2.53) days; postoperative time back to work was (29.40 ± 8.75) days; all differences of the results were significant between two groups (P < 0.05).According to function of ulner nerve scoring system,one year postoperatively, excellent or good results were 82.14% in routine group and 81.25% in endoscope group,no significant difference between two groups (P > 0.05). Conclusion Compared with routine anterior transposition of the ulnar nerve,endoscopically assisted anterior transposition has the following advantages: smaller incision and less tissue damage,less postoperative pain and sooner returning to work.And similar outcome was achieved from the two group.
6.Clinical application of finger soft tissue defect coverage with dosal proximal digit fatty fasciocutaneous flap
Feiya ZHOU ; Liangfu JIANG ; Yonghuan SONG ; Jinquan YANG ; Jian DING ; Tinggang CHU
Chinese Journal of Microsurgery 2016;39(4):324-327
Objective To discuss the coverage of finger soft tissue defect with dorsal proximal digit fasciocutaneous flap on the middle and distal digit.Methods From May,2013 to December,2014,8 cases with soft tissue defects at 8 fingers were treated with dorsal proximal digit fasciocutaneous flap.The flap sizes ranged from 2.5 cm × 2.0 cm to 3.5 cm × 3.0 cm.The donor site were closed straightly.Results Eight flaps of 8 fingers survived.All the wounds at the donor sites healed well.Eight fingers in 8 cases were followed up for 6-12 months.The color,texture and contour of the flaps were satisfied.The two-point discrimination distances were 8-10 mm.Conclusion The skin defect in the middle and distal digit can be satisfied covered with dorsal proximal digit fasciocutaneous flap.This flap is a simple,reliable and safe management for digit defect and can be performed in the primary hospital.To ensure the surviving of the flap,ensure the surviving of the flap,the awareness of the anatomy of the flap should be known well.The limits of its reconstruction of sensation and coverage size exit in its application.
7.Association between the Apolipoprotein E gene polymorphism and traumatic brain injury
Yi GU ; Xingjie GAO ; Tao XU ; Gan WANG ; Jin HU ; Bhattarai BINOD ; Dongxiang WANG ; Sanduo JIANG ; Liangfu ZHOU
Chinese Journal of Nervous and Mental Diseases 2007;33(7):385-388
Background To explore the relationship between polymorphism of APOE gene in traumatic brain injury(TBI)patients suffering from traffic accident and the outcome of TBI.Methods TBI patients were randomly selected in this study with caxe-wntrol trial. The genotype of APOE allele was tested by a polymerase chain reaction-restriction fragment length polymorphism ( PCR-RFLP), and the association between different genotypes of APOE alleles and outcome of TBI patients, were analyed.Results In TBI group frequency of APOE ε2 allele was 0. 1010, and frequency of APOE ε2/ε3 was 0. 1596.Both of these results were significantly higher than that in normal people (APOE epsilon 2 was 0. 0050, APOE ε2/ε3 was 0. 0100) (P<0.05). Frequency of APOE ε2 and APOE ε2/ε3 in TBI group who died was 0. 1970 and 0. 2727. These were significantly high compared to TBI patients who had good recovery.Conclusions APOE allele ε2 and APOE genotype ε2/ε3alleles indicate a poor prognosis of traumatic brain injury patients.
8.Repairation of composite distal soft tissue defect of thumb and finger with mini toenail flap
Feiya ZHOU ; Liangfu JIANG ; Xian ZHANG ; Weiyang GAO ; Yonghuan SONG ; Jian DING ; Tinggang CHU
Chinese Journal of Microsurgery 2019;42(4):322-325
To introduce the repairation procedure of composite distal soft tissue defect of thumb and finger with mini toenail flap. Methods From June, 2015 to June, 2018, 7 cases with composite tissue defect at 7 distal fingers, including 5 thumbs, 1 index finger and 1 middle finger, were reconstructed with mini toenail flap transfer.The flap sizes which were raised during the operation ranged from 4.5 cm×3.0 cm-3.0 cm×1.5 cm.The donor sites were covered by toe phalanx shortening, V-Y advancement flap and local pedicle flap. Microsurgical routine treatment was made after the operation, and followed-up regularly. Results Seven flaps of 7 cases completely sur-vived without any necrosis. All the wounds at the donor sites healed well. All patients were followed-up for 6-36 months. The motive, sensor and cosmetic result were satisfied. In sensory function, the two-point discrimination dis-tance could restore to be 4-6 mm. Conclusion The mini toenail flap transfer is a reliable and suggested method.It can anatomically restored the distal digit sensor function with cosmetic contour, and regain the motive, sensory func-tion and satisfied cosmetic appearance.
9. Treatment of the defect on digit and toe with modified Reading man flap
Tinggang CHU ; Feiya ZHOU ; Jian DING ; Anyuan WANG ; Zengming ZHENG ; Liangfu JIANG ; Hede YAN ; Xinglong CHEN
Chinese Journal of Plastic Surgery 2019;35(10):1000-1003
Objective:
To introduce the design of modified Reading man flap, and observe the clinical effect of modified flap for repairing digit and toe wounds.
Methods:
From July 2014 to September 2017, dorsal skin defects on 37 digits and 18 toes were repaired with our modified Reading man flaps. The revised design was characterized with enlargement the major flap and extending the minor flap proximally, and all donor site defect were sutured primarily through the major flap recover the defect and the minor flap repair the subsequent donor defect.
Results:
All the detects in 55 patients were repaired by the modified Reading man flaps with direct donor sites closure. With average of 11.5 months (9.5-25.7 months) follow-up, all flaps survived with satisfactory texture and appearance, the bulky deformity and scar contracture did not occur. Partial necrosis of tip in the minor flaps occurred in 2 toes and healed by wound dressing. The function of the toe joints was good and the walking gait was normal. Partially impaired PIP joint function with limited flexion occurred in 2 cases. Based on the TAM evaluation criteria, the results were excellent in 28 digits, good in 7 digits, and the overall satisfactory rate was 94.6%.
Conclusions
The modified Reading man flap can get good clinical effects for treatment of the digit and toe dorsal skin defect with the advantages of simple procedure, easy transfer and direct closure of donor sites. Flaps appearance and joints function can get good result postoperatively.