AIM: To investigate the effect of mfn2 on mitochondrial function in steatosis hepatocytes. METHODS: Plasmid pEGFP-mfn2 was transfected into hepatocyte strain L02 by Lipofectamine 2000 in vitro, then the steatosis model of hepatocytes was establish by oleic acid induction. RT-PCR was used to evaluate mRNA expression and Western blotting was use to detect the protein expression. ATP level was determined by firefly luciferase bioluminescent. ROS production was measured by fluorescence probe DCFH-DA. Chondrosome transmembrane potential of L02 was observed by labeling of JC-1 and FCM. RESULTS: The stable expression of ectogenesis mitofusin2 in L02 cells was confirmed by RT-PCR and Western blotting. In the model of oleic acids induced lipid formation, Mfn2 obviously inhibited the descent of chondrosome transmembrane potential and ATP level, and increased ROS production in L02 cells. CONCLUSION: Up-regulated expression of mfn2 attenuates mitochondria dysfunction caused by oleic acids induced lipid formation.

Objective To evaluate modified retroperitoneal laparoscopic resection of renal cyst. Method Thirty-six patients with renal cyst were treated by modified retroperitoneal laparoscopic resection of renal cyst,summarized the clinic data and follow-up the effect. Results All 36 cases were operated suc-cessfuUy without changing to opening operation,average operation time (50 ± 35)min,no complications oc-curred and no recurrence was found. Conclusions The modified retroperitoneal laparoscopic resection of renal cyst with two 5 mm-trocars and one 10 mm-trocar has less trauma than classic laparoscopic operation. It is one of mini-trauma operation method which is worth to be popularized in clinic.

Objective To explore the application ways of IHE XDS technology in electronic medical record (EMR) system and develop application system of clinical EMR integration. Methods The clinical EMR integration plan was worked out based on IHE XDS technical framework. Results A EMR solution based on IHE XDS was presented. It was consisted of the components of the clinical Data Entry, XDS Registry, clinical document Repository, EMR application server and the i-node accessing server. The system was tested and the result indicated that the system was effective and scalable. Conclusion IHE XDS profile can be not only used to realize the sharing of EMR documents across multiple hospital, but also support the collection clinical data within a hospital clinical system integration.

ObjectiveTo examine the effects of proprioceptive neuromuscular facilitation(PNF) techniques on lower limb motor function of strokes and its mechanism.MethodsWe used simple random sampling and cross-section survey design. PNF Contract Relax Agonist Contract(PNF-CRAC) techniques were applied to 44 stroke patients. Surface electromyography values(sEMG) was recorded from rectus femoris and hamstring of stroke patients with both low limbs. ResultsPNF-CRAC techniques not only caused the irradiation of muscles activities in the contralateral extremity during unilateral exercise, but also increased agonist EMG activities and the motor control of knee joint in strokes.ConclusionPNF-CRAC techniques can improve the knee stability and enhance the recovery of motor function in paretic lower limb.

This review summarized the research status on DNA molecular identification of plants in the genus Chrysanthemum. Some case studies on representative DNA molecular identification techniques, which included ge-nomic in situ hybridization (GISH), DNA molecular markers and DNA barcoding, were described. Simultaneously, the merits, demerits and development of the techniques were discussed. The above work provides evidence for the identi-fication and resource utilization of plants in the genus Chrysanthemum.

Aim Toinvestigatewhetherexposureto Sanguinarine (SAN ) can inhibit cell proliferation in human mammary adenocarcinoma cells (MCF-7 ) and thepossiblemechanism.Methods WeexposedMCF-7 to anticancer compound SAN,cell viability was as-sessed by using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT ) reduction assay. ROS was measured using confocal microscopy,expres-sion of caspase-3 ,caspase-8 and caspase-9 were calcu-latedusingchemiluminescencemethod.Results SAN remarkably inhibited growth of human mammary adeno-carcinoma MCF-7 cells by decreasing cell proliferation. ROS release and caspase-3,caspase-8,caspase-9 ex-pression were stimulated by SAN in MCF-7 ,and these changes were abolished by the antioxidant,N-acetyl-cysteine(NAC).Conclusion Regulationofcaspases expression and release from MCF-7 cells are possibly e-voked by SAN through reactive oxygen species.

Objective To investigate the effect and mechanism of HPV16-E7 gene specific CRISPR/Cas (clustered regularly interspaced short palindromic repeat/CRISPR associated system) on the cell apoptosis and proliferation of human cervical cancer SiHa cell line.Methods HPV16 positive cervical cancer SiHa cells and HPV16 negative human embryonic kidney HEK293 cells were cultured and each of the cells were divided into 3 groups respectively in the experiment:the control group was untreated,the C9 group was transfected only with Cas9 plasmid,and the Cri group was cotransfected with guide RNA (gRNA) plasmid and Cas9 plasmid (1∶3).T-7endonuclease Ⅰ assay was used to detect double strand break (DSB) formation in SiHa cells.The apoptosis rates of SiHa and HEK293 cells were detected by flow cytometry (FCM).CCK-8 was used to evaluate the proliferation of SiHa and HEK293 cells.Western blotting was applied to detect E7 and pRb protein expression.Results The SiHa cells in Cri group showed DSB formation at 48 h after transfection.The apoptosis rate of Cri group at 48 h was 26.6％,higher than 2.6％ in control group (x2 =5.455,P =0.020) and 3.1％ in C9 group (x2 =6.279,P =0.012).The apoptosis rates of control,C9 and Cri group were 7.4％,7.6％,7.9％ for HEK293 cells respectively.Compared with the control and C9 group,the apoptosis rate of the Crigroup showed no statistical significance (x2 =0.032,P =0.858;x2 =0.034,P =0.853).After 72 h transfection,the inhibition rate of SiHa cells in Cri group was 29.4％,higher than 15.0％ in control group (x2 =22.481,P =0.000) and 18.0％ in C9 group (x2 =24.879,P =0.000).The inhibition rate of the HEK293 cells in Cri group was 3.2％,and it showed no significant difference compared with the inhibition rate of C9 group (2.2％,x2 =2.857,P =0.091) and control group (2.3％,x2 =3.438,P =0.064) respectively.Down-regulated expression of E7 protein and up-regulated expression of pRb protein were detected of the SiHa cells in Cri group compared with the control group,while the E7 protein and pRb protein level did not show difference in the C9 group.Conclusion CRISPR specifically targeting HPV16-E7 oncogene can promote apoptosis of SiHa cells,and inhibit the cell proliferation.It is speculated that CRISPR system may induce DSB event of E7 gene,and result into increased expression of tumor suppressor protein pRb.

Based on the competition reaction of target protein, aptamer probe, padlock probe and complementary sequence, a highly sensitive fluorescent aptasensor was developed in this study in combination with rolling circle amplification. In the absence of target protein, the ligation-rolling circle amplification reaction was repressed because the complementary sequence hybridized with aptamer probe to form double-stranded duplex. While in the presence of target protein, the target molecules bound specifically with aptamer probe, inducing displacement of the complementary sequence and hybridization with padlock probe. The padlock probe was circularized with the assistance of E. coli DNA ligase, and the rolling circle amplification process could be accomplished by Phi 29 DNA polymerase. The amplification product contained thousands of repeated sequences which could hybridize with the loop of molecular beacon ( the detection probes) , resulting in a significant fluorescence signal. The effects of length of complementary DNA ( CDNA ) sequence and concentration of padlock probe were investigated. Under the optimized experimental conditions, the model target protein thrombin could be highly sensitively detected by the proposed aptasensing system in a linear range of 0 . 067-32 . 4 nmol/L with a detection limit of 0 . 03 nmol/L ( approximately 90 amol target molecules). Moreover, the presented sensing method was universal for other target analysis by skillfully design of the sequence of aptamer probe and related oligonucleotides.

This study on determination of leukemia-specific chromosomal abnormalities and their relationship with prognosis of childhood acute leukemia (AL) had an important significance for childhood acute leukemia. In recent years, the efficacy of treatment of childhood AL has been greatly improved, but relapse is still a main factor affecting prognosis. Treatment based on the risk stratification by cytogenetic abnormalities can improve the prognosis and survival rate. In the past 3 decades, the genetic techniques have developed rapidly and many new genetic abnormalities have been found. This review highlights the main chromosomal and genomic abnormalities of 3 common childhood AL, including B-cell precursor acute lymphoblastic leukemia (BCP-ALL), T-cell acute lymphoblastic leukemia (T-ALL) and acute myeloid leukemia (AML).
Acute Disease ; Child ; Humans ; Leukemia ; genetics ; Leukemia, Myeloid, Acute ; genetics ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; genetics

Objective:To investigate the expression and clinical significance of wild-type p53-induced phosphatase 1 (Wip1) in thyroid carcinoma and biological effect of siRNA-targeting Wip1 on the thyroid carcinoma cell line. Methods:Immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) were performed to detect the expression of Wip1 in 73 specimens of thy-roid carcinoma tissues and normal thyroid tissues (5 cm away from the margin of thyroid carcinoma), respectively. Wip1 siRNA was transiently transfected into the papillary thyroid carcinoma cell by using a liposome-mediated method and then detected by RT-PCR and Western blot. Methyl thiazolyl tetrazolium (MTT) assay and flow cytometry (FCM) were also conducted to observe cell proliferation, cell apoptosis, and cell cycle. Results:The positive rates of Wip1 protein were 80.8%in thyroid carcinoma tissues and 9.6%in the nor-mal tissues (χ2=47.036, P<0.05). The relative mRNA contents of Wip1 were 0.665 ± 0.046 and 0.225 ± 0.039 in carcinoma and normal tissues, respectively;these results significantly differed between the two types (t=12.637, P<0.05). Significant correlation was not ob-served between Wip1 expression and other factors, such as patient's gender, age, and tumor size (P>0.05). However, significant correla-tions among Wip1 expression, lymph node metastasis, clinical stages and tumor differentiation (P<0.05) were observed. RT-PCR and Western blot results showed that K1 cell-transfected Wip1 siRNA exhibited a relatively lower expression than normal cells (t=17.039, t=14.637, P<0.05). MTT assay results showed that the K1 cells transfected with Wip1 siRNA showed a lower survival fraction, higher cell apoptosis, higher percentage of G0/G1 phases, and lower cell concentration in G2/M and S phases (P<0.05). Conclusion:Wip1 pro-tein and mRNA were increased in thyroid carcinoma and are correlated with lymph node metastasis, clinical stages and tumor differenti-ation. Wip1 may be involved in proliferation, apoptosis, and cycle of thyroid cancer cells.