1.Effect of different wave lengths of low power laser on fracture healing
Ye MENG ; Minghao JIN ; Ruifeng GE
Chinese Journal of Tissue Engineering Research 2009;13(28):5421-5424
BACKGROUND: Low power laser irradiation has positive effects on fracture healing, including shortening the time of bone union and enhancement of bone formation. OBJECTIVE: To evaluate the effects of different wave lengths in low power laser on eedy fracture healing. DESIGN, TIME AND SETTING: Randomized, controlled, animal experiment. The study was performed at the Laser Center of Dankook University Medical Center between May 2005 and May 2006. MATERIALS: A total of 60 female English Hartley guinea pigs, 9 weeks old, weighing (300~30) g, were used for this experiment. They were randomly divided into three groups control, 632, and 830 nm groups (n=20). METHODS: The right femoral middle shaft fractures were made with a bone cutter and fixed with intramedullary nails (1.4 mm diameter K-wire). The frequencies of 632 nm and 830 nm semi-conductor laser machines were used. The laser irradiations were started 48 hours after operation vertically at the fracture sites and were applied every two days, at 100 J/cm<'2> of the same irradiation doses and 2.52 cm<'2> of the same irradiation areas in different irradiation groups. Finally, 46 guinea pigs survived, including 14 in control group, 16 in 632 nm group and 16 in 830 nm group. MAIN OUTCOME MEASURES: Guinea pigs were sacdficed at 3 and 6 weeks separately. The bone healing was assessed by the Modified Zorlu Scoring System, including gross, radiologic and histologic examinations. RESULTS: In the gross and radiologic findings, both 632 nm and 830 nm irradiation groups shown a significantly greater rate of callus formation at postoperatively 3 weeks compared to the control group (P< 0.01). Quantity of callus formation in 830 nm group was more than 632 nm group, but there was no significant difference (P > 0.05). The result of histological findings showed a significant increase of osteoblastic proliferation in two irradiation groups than the control group at 3 weeks postoperatively (P < 0.01). Enhancement of osteoblastic proliferation was more obvious in 830 nm group than in 632 nm group at 3 and 6 weeks, but there was no significant difference (P > 0.05).CONCLUSION: Both 632 nm and 830 nm wave lengths are considered to be optimal irradiation wave lengths for eady fracture healing. Moreover, 830 nm irradiation is better than 632 nm irradiation for eady fracture healing.
2.Intervention of Ciliary Neurotrophic Factor on Weightlessness-Induced Muscle Atrophy and Myofiber Phenotype Transition
Peng ZHANG ; Hongju LIU ; Minghao YANG ; Li LI ; Xin GE ; Xiang LI ; Xu WANG ; Ming FAN ; Xiaoping CHEN
Space Medicine & Medical Engineering 2008;21(3):240-244
Objective To investigate the intervention of ciliary enurotrophic factors(CNTF)on weightlessness-induced muscle atrophy and its myofiber phenotype transition.Methods Tail-suspended (hindlimb unloading)of adult male Wistar rats were used to create the mode J of weightlessness-induced muscle atrophy.The effect of CNTF treatment on weightlessness-induced muscle atrophy and its myofiber phenotype transition was determined by analyzing the expression changes of MHCI/IIb and p130 or Myf5 with RT-PCR and Western blot.Results CNTF treatment in vivo markedly reversed weightlessness-induced muscle weight IOSS selectively in the slow-twitch muscle soleus.Moreover,during tail suspension,soleus weight loss and myofiber phenotype transition indicated that CNTF treatment could sigificantly attenuate the weight loss and slow to fast myofiber phenotrype transition in soleus compared with control(CNTF untreated and tail suspended).Furthermore,it was showed that the CNTF-induced intervention effect was associated with the protein level upregulation of muscle satellite cell-specific markers,P130 and Myf5.The satellite cell pool in CNTFtreated soleus was increased.Conclusion It iS firstly demonstrated that CNTF can attenuate weightlessness-induced muscle atrophy and its myofiber phenotype transition to be through the increase of satellite cell pool in soleus.
3.Expression of silent information regulator 1 and 3 and hypoxia-inducible factor 1α in cutaneous squamous cell carcinoma tissues and cells
Xinhong GE ; Yaning JIAO ; Minghao GE ; Yingdong MA ; Yue SHI ; Yu WANG ; Lingling LIU
Chinese Journal of Dermatology 2022;55(2):116-122
Objective:To determine the expression of silent information regulator 1 (Sirt1) , Sirt3 and hypoxia-inducible factor 1α (HIF-1α) in cutaneous squamous cell carcinoma (CSCC) tissues and cells, and to explore their role in the occurrence and development of CSCC.Methods:From January 2019 to December 2020, 30 lesional skin tissues were obtained from patients with histopathologically confirmed poorly-, moderately- or well-differentiated CSCC, and 30 normal skin tissues were obtained from patients with non-cancerous diseases in Department of Dermatology, General Hospital of Ningxia Medical University. A CSCC cell line A431 and a human keratinocyte cell line HaCaT were cultured. Immunohistochemical study, Western blot analysis and real-time quantitative PCR (RT-PCR) were performed to determine the protein and mRNA expression of Sirt1, Sirt3 and HIF-1α in CSCC tissues of different grades of differentiation and normal skin tissues, cytochemical and immunofluorescence staining and RT-PCR were conducted to determine the protein and mRNA expression of Sirt1, Sirt3 and HIF-1α in A431 and HaCaT cells, respectively. Comparisons of measurement data among multiple groups were performed by using one-way analysis of variance, and comparisons between two groups by using t test. Results:Immunohistochemical study showed that the expression level of Sirt3 (expressed as the average optical density) was 100 ± 12.12, 117.72 ± 26.23, 127.32 ± 24.45, 132.71 ± 31.61 in the normal skin tissues and well-, moderately- and poorly-differentiated CSCC tissues respectively, and there was a significant difference among these groups ( F = 20.14, P < 0.001) ; the expression of Sirt1 and HIF-1α increased in turn from the normal skin tissues to the well-, moderately- and poorly-differentiated CSCC tissues, and significantly differred in these groups ( F = 174.50, 225.00, respectively, both P < 0.001) . As Western blot analysis revealed, the expression level of Sirt3 significantly differed among the normal skin tissues, well-, moderately- and poorly-differentiated CSCC tissues (expressed as relative gray value: 1.000 ± 0.132, 1.403 ± 0.411, 1.387 ± 0.393, 1.677 ± 0.683, respectively; F = 34.97, P < 0.001) , and so did the expression levels of Sirt1 and HIF-1α ( F = 69.29, 199.90, respectively, both P < 0.00l) , with a gradually increasing trend in their expression levels from the the normal skin tissues to well-, moderately- and poorly-differentiated CSCC tissues. RT-PCR showed that the mRNA expression of Sirt3, Sirt1 and HIF-1α was sequentially increased from the normal skin tissues to well-, moderately- and poorly-differentiated CSCC tissues, and significant differences were observed among these groups ( F = 113.00, 174.50, 50.33, respectively, all P < 0.001) . The protein expression levels of Sirt3, Sirt1 and HIF-1α were significantly higher in the A431 cells than in the HaCaT cells ( t = 16.75, 18.34, 27.76, respectively, all P < 0.001) , and so were their mRNA expression levels ( t= 14.22, 9.62, 16.86, respectively, all P < 0.001) . Conclusion:Increased expression of Sirt3, Sirt1 and HIF-1α was observed in CSCC tissues and cells, which may promote the occurrence and development of CSCC.
4. Rapid detection of human adenovirus by recombinase polymerase amplification assay and lateral flow dipstick
Kangchen ZHAO ; Yiyue GE ; Lunbiao CUI ; Yin CHENG ; Zhiyang SHI ; Fengcai ZHU ; Minghao ZHOU
Chinese Journal of Experimental and Clinical Virology 2017;31(4):357-361
Objective:
To establish a rapid and sensitive isothermal amplification assay for the detection of human Adenovirus.
Methods:
Primers and probe used for recombinase polymerase amplification(RPA)were designed based on the conserved region of the adenoviruses hexon gene. After optimizing the reaction temperature and times, the products of RPA were detected by capillary electrophoresis and lateral flow dipstick(LFD). Sensitivity and specicity of the assay were evaluated. The diagnostic value of the RPA-LFD assay was verified using clinical samples which were simultaneously tested by real time PCR assay.
Results:
The analytical sensitivity of RPA-LFD assay was 2 copies DNA molecules per reaction and no cross reaction with other pathogens was observed. Compared with real-time PCR assay, the sensitivity, and specificity of the present assay were all 100%.
Conclusions
The RPA-LFD assay developed in this study has the characteristics of high specificity, sensitivity, rapid and no requirement of expensive equipment which provided a new tool for rapid detection of human adenovirus.