Objective To investigate the prevalence of mtDNA A3243G mutation in the early onset diabetics in Tianjin for exploring the relationship between mtDNA mutation and diabetes. Methods 348 kinship-free diabetics whose ages at onset were less than 45y were randomly recruited, with 207 control subjects. The PCR-RFLP and cloning techniques were applied to screen the A to G substitution at nucleotide 3243 of mtDNA tRNALEU（UUR）. Meanwhile, the clinical and genetic analysis was done from one pedigree. Results We detected two diabetics harboring the well-known a3243g mutation with the mutation frequency of 0.6%. While the mutation frequency in diabetics with （positive） family history was 1.2%, and zero in the control subject. The proband showed a typical （picture） of mitochondrial diabetes mellitus, while the other family members harboring the same （mutation） had heterogeneous presentation. Conclusion The prevalence of mtDNA A3243G mutation in early onset diabetes of Tianjin is low, but relatively higher in diabetes complicated with other （symptoms） of mitochondrial disease. Furthermore, its heterogeneity would decrease with aging in （mitotic） tissue

Objective To investigate clinical traits of 2 families members habouring mtDNA 12026A→G mutation based on our previous studies.Methods 25 members in 2 families with probands with mtDNA 12026 mutation were examined.All their clinical and biochemical data were collected.Total genome was extracted conventionally from peripheral leucocytes of all participants,and PCR-RFLP techniques were applied to screen A to G substitution at nucleotide 12026 of mtDNA in ND4 region.Results We found 13 individuals habouring the 12026 A→G mutation in 2 pedigrees,all without deafness.Among them,5 with diabetes were found.Interestingly,we found 3 individuals with hyperthyroidism in one family(one also combined with diabetes).Conclusions Our findings suggest that diabetic families with mtDNA 12026 A→G mutation in ND4 region can have different clinical pictures,and may involve in autoimmune diseases

Objective To investigate clinical traits of 2 families members habouring mtDNA 12026A→G mutation based on our previous studies.Methods 25 members in 2 families with probands with mtDNA 12026 mutation were examined.All their clinical and biochemical data were collected.Total genome was extracted conventionally from peripheral leucocytes of all participants,and PCR-RFLP techniques were applied to screen A to G substitution at nucleotide 12026 of mtDNA in ND4 region.Results We found 13 individuals habouring the 12026 A→G mutation in 2 pedigrees,all without deafness.Among them,5 with diabetes were found.Interestingly,we found 3 individuals with hyperthyroidism in one family(one also combined with diabetes).Conclusions Our findings suggest that diabetic families with mtDNA 12026 A→G mutation in ND4 region can have different clinical pictures,and may involve in autoimmune diseases.

Objective To investigate the relationship between K+ concentration in rabbit vitreous humor and postmortem interval (PMI) under different ambient temperature. Methods Rabbit corpses were stored at 5℃ , 15℃ , 25℃ , and 35℃ after execution, and 80~100μL vitreous humor was extracted from each eye of the rabbit in turn every 12 hours. The concentrations of K+ were examined by Modular DPPI automatic biochemistry analyzer. The Interpolation Functions were used to analyze the statistical relationship between PMI and K+ concentration under different temperature. Results In each animal group, K+ concentration increased with PMI. Equation was obtained after interpolation analysis on range of temperature 5℃ ~30℃ . The three-variable quintic surface equation was f(x,y)=-1.998e14+1.345e12x+5.902e13y+0.005585x2-4.509e11xy-3.876e12y2-0.0002868x3+0.003545x2y+4.406e10xy2-1.746e10y3+2.669e-6x4-1.568e-5x3y-0.0001771x2y2-1.64e9xy3+6.669e9y4-8.672e-9x5+4.467e-8x4y+2.354e-7x3y2+2.459e-6x2y3+2.05e7xy4-1.214e8y5(R2=0.9956), x stands for temperature, y stands for K+ concentration, f(x,y) stands for PMI. Conclusion The rule of K+ concentration changes at ambient temperature complied with three-variable quintic surface equation distribution. Measurement of interpolation function may be used for PMI estimation at different ambient temperature.

Background and purpose:Low intensity ultrasound (LIUS) can kill cancer cells and promote their apoptosis. However, it is still unknown how it affects the migration and invasion of tumor cells. This study aimed to explore the effect of LIUS on human hepatocellular line MHCC97H in migration and metastasis and the possible mechanismin vitro.Methods:According to the intensity of ultrasonic irradiation, 4 experimental groups were established: control group (0 W/cm2), 0.5 W/cm2, 1.0 W/cm2 and 1.5 W/cm2group. The migration and invasion ability of hepatocellular cells was detected by scratch assay and Transwell migration and invasion assay after the irradiation of LIUS. The changes of cytoskeleton after irradiation were observed by microscope and F-actin green lfuorescence staining. The expressions of MMP-2 and MMP-9 were examined by real-time lfuorescent quantitative polymerase chain reaction (RTFQ-PCR) and Western blot.Results:Low intensity ultrasound (≤1.5 W/cm2) promoted the migration and invasion of hepatocellular line MHCC97H. Scratch assay and Transwell assay showed much more cells under irradiation migrated through membrane than untreated. It was found that morphology of liver cancer cells changed after LIUS irradiation using optical microscope and lfuorescence microscope. The results of RTFQ-PCR and Western blot showed upregulation of MMP-2 expression by LIUS in MHCC97H and high expression of MMP-9 mRNA. Conclusion:Low intensity ultrasound may promote the migration and invasion of MHCC97H through changing cytoskeleton and upregulating protein expression of MMP-2.

Objective To explore whether the low-intensity pulsed ultrasound (LIPU) could induce apoptosis on SMMC-7721 cells and to explore the underlying mechanism.Methods The SMMC-7721 cells were randomly divided into 4 groups:a blank control group,which was subject to sham exposure to ultrasound,and 3 ultrasound intervention groups exposed to ultrasound at intensities of 0.5,1.3 and 2.0 W/cm2,respectively.Then they were incubated for 6 h.The cell apoptosis,necrosis and changes of cell cycles were measured using the flow cytometry.The transmission electron microscope (TEM) was used to observe microstructural changes in the cells.The agarose gel electrophoresis (AGE) was used to examine the DNA fragmentation,and Western-blotting was employed to assess the protein expression of caspase-3.Results The average cell apoptosis rate of the 3 intervention groups were 4.66％,8.99％ and 32.41％,respectively.The percentage of cells in G2 phase increased significantly and those in G1 phase decreased significantly in the 3 intervention groups compared to the blank control group at the same time points.In the intervention groups,significant cell apoptosis was observed under TEM,and DNA ladders was seen in AGE,with DNA fragments appearing obviously when cells were incubated for 6 h and 9 h after ultrasound exposure.In intervention groups subject to 1.3 and 2.0 W/cm2 ultrasound exposure,the protein expression of caspase-3 was significantly higher than that of the control group.Conclusion LIPU can inhibit the proliferation and induce apoptosis of SMMC-7721 cells with a dose-dependent feature.The possible mechanism underlying the LIPU-induced cell apoptosis might be related to the activation of the mitochondria pathway,and especially the caspase-3 protein.

BACKGROUND: Iliac bones are common habitant parts in metastatic tumor of bone, and the affiliate cortex is easy to be penetrated into soft tissue blots. The irregular iliac shape and its complex surrounding tissues create the difficulties in puncture operations and cement perfusion.OBJECTIVE: To investigate the clinical outcomes of CT-guided versus DSA-guided puncture combined with bone cement perfusion for metastatic tumor to the ilium.METHODS: Sixty-four patients with metastatic tumor to the ilium udergoing bone cement perfusion were enrolled, and then divided into control and experimental groups (n=32 per group), followed by treated with DSA-guided percutaneous injection of bone cement, and CT-guided puncture, respectively. Subsequently, the diffuse distribution of bone cement was observed using CT. The success rate, leakage rate, Visual Analogue Scale scores and Oswestry Disability Index were compared between two groups.RESULTS AND CONCLUSION: (1) All patients received the surgery successfully, and bone cement leakage happened in two cases of control group and one case of experimental group. (1) The Visual Analog Scale and Oswestry Disability Index scores in the two groups were significantly better than those before treatment (P < 0.05), and the scores showed no significant differences between two groups (P > 0.05). (3) No difference was found in the pain relief and functional recovery between two groups. (4) To conclude, CT-guided treatment can achieve satisfactory outcomes for metastatic tumor to the ilium by bone cement infusion in appropriate time.

The active copper-containing carbon nanodots were prepared by hydrothermal method, and then characterized by fluorescence spectroscopy and UV-visible absorption spectroscopy.Subsequently, a highly sensitive and selective electrochemical biosensor was fabricated on the basis of this synthesized carbon nanodots with electro-deposition technique.The electrode behavior was investigated by cyclic voltammetry, electrochemical impedance spectroscopy, and differential pulse voltammetry.Furthermore, the catalysis mechanism was studied.The experimental results indicated that the biosensor exhibited a strong electrocatalytic activity toward the oxidation of uric acid (UA).What′s more, the interference from ascorbic acid and dopamine was eliminated effectively.Under the optimum conditions, there were linear relationships between the anodic peak current and the concentration of UA (1.00-300.0 μmol/L), and the limit detection was 0.30 μmol/L (S/N=3).The prepared biosensor had advantages such as easy fabrication, strong anti-interference ability, high sensitivity, and wide detection range, and could be used for real sample detection.

Objective To investigate the risk factors of severe hand,foot and mouth disease (HFMD).Methods 175 severe cases of HFMD and 183 mild cases of HFMD in the same period were randomly selected.Single factor analysis was first performed between severe and mild cases on age,gender,residence,symptoms,signs and laboratory examinations,etc,to screen out the related risk factors which P value ＜ 0.05.Then,binary logistic regression analysis was carried out to determine risk factors most related to severe HFMD.Finally,receiver operating characteristic curve (ROC) analysis was performed on severe HFMD related risk factors.Results Single factor analysis showed that there were obvious differences between children with mild HFMD and those with severe HFMD in the factors like difficulty in breathing,walking instability,vomiting,limb shaking,disturbance of consciousness,convulsions,cold sweat and weakness,thermal process,the degree of fever,pulmonary rales,heart rate,serum EV71 antibody,circulatory failure,leukocyte count,platelet count,neutrophil ratio,CRP,blood glucose,etc (x2 =15.236,19.819,33.823,52.670,12.984,10.180,29.318,52.932,34.544,14.615,46.633,31.407 and 5.303,t =3.184,3.144,2.256,2.244 and 2.828,,all P ＜0.05).Binary logistic regression analysis showed that the thermal process,startle tremor or limb jitter,serum EV71 antibody,vomiting,fever,neutrophil ratio were the related risk factors of severe HFMD (B value =2.605,2.129,1.409,1.185,0.841 and 0.103,all P ＜ 0.05).ROC analysis showed that the areas under the curve of the predicted probability and thermal process were larger than any other risk factors [(95％ CI (0.888 ～ 0.961) and (0.818 ～ 0.920)],and thus had better diagnostic values.Conclusion Children under 3 years old were the high risk population of HFMD.Such clinical symptoms as persistent high fever,vomiting,startle tremor orlimb jitter,EV71 antibody in serum and increasing neutrophil ratio were risk factors for severe HFMD.The predicted probability had more diagnostic value than any other risk factors.

Objective: To explore the repair methods and effects of the complex wounds on hands after burns or trauma. Methods: From January 2008 to December 2017, 45 patients (28 males and 17 females, aging 8 to 58 years) with severe hand injuries after burns or trauma combined with deep tissue exposure were admitted to our hospital. Two patients had whole-fingers degloving injuries, 27 patients had dorsal hand injuries, and 16 patients had palmar injuries. After debridement, the area of soft tissue defects was 7 cm×6 cm to 19 cm×12 cm combined with 0.5 cm×0.4 cm to 10.0 cm×4.0 cm of single deep tissue exposure. Different repairing methods were adopted according to the area and location of deep tissue exposure. Five patients with small area exposure were treated with artificial dermis+ vacuum sealing drainage (VSD)+ autogenous skin grafting. Thirty-eight patients with unilateral large area exposure on palm or dorsum were treated with segmented ligation of abdominal thin flaps (with area of 8 cm×7 cm to 15 cm×9 cm). Two patients with bilateral large area exposure in dorsal and palmar hands were treated with modified abdominal bag-shaped delayed thin flaps (with area of 12 cm×5 cm to 12 cm×9 cm and 12 cm×6 cm to 14 cm×9 cm). The donor sites were directly sutured or repaired with intermediate split-thickness skin or adjacent flap. The survival of grafts and flaps was observed, number of operations, wound healing time, and follow-up were recorded. Results: (1) Among the patients receiving artificial dermis+ VSD+ autogenous skin grafting, the wounds of 3 patients were healed after 2 operations, and 2 patients had artificial dermis infection and lysis, and tendon necrosis, which were healed after 3 operations. The wound healing time of 5 patients was 14 to 33 days post injury. During the follow-up of 3 months, the affected hands were in good shape with soft texture and fewer scars, and functional evaluation of hand was good in 3 cases and modest in 2 cases. (2) The patients receiving segmented ligation of abdominal thin flaps all underwent 2 operations without flap necrosis. The wound healing time was 2 to 3 weeks post injury. Thirty-five patients underwent one to six-years′ follow-up, which showed that the flaps were in good shape and color with soft texture, and the functional evaluation of hand was excellent in 25 cases, good in 7 cases, and modest in 3 cases. Three patients were lost to follow-up. (3) In the 2 patients receiving modified abdominal bag-shaped delayed thin flaps, all flaps survived after 5 operations, the wounds were healed on post injury day 22 and 24 respectively, the shape and texture of the affected hands was good with no bloated appearance after separating fingers and revision. During the follow-up of 2 years, the functional evaluation of hand was good in 2 cases. Conclusions For the complex wounds on hands after burns or trauma, if the area of deep tissue exposure is small, artificial dermis+ VSD+ autogenous skin grafting should be adopted, which has good effects. If the area of unilateral deep tissue exposure is large, segmented ligation of abdominal thin flap should be adopted; if the area of bilateral deep tissue exposure is large, modified abdominal bag-shaped delayed thin flap should be adopted. These methods can reduce the number of operations, shorten wound healing time, and obtain good shape and function of hands.