Objective To analyze the chara cteristic finding of metachromatic leukodystrophy (MLD) on MR images. Methods Eleven patients with biochemicall y proved MLD were studied retrospectively for the extent of white matter abnormalities and the in volvement of specific structures (such as corpus callosum, white matter tract, a nd thalami). Result All 11 cases in the periventricular white m atter and 10 cases in centrum semiovale showed a symmetric confluent high intens ity on T_2-weighted images. Involvement of subcortical U-fibers was present i n 4 late-stage cases. Other sites of involvement were the genu (n=8) and s plenium (n=9) of the cor pus callosum (both n=8), the posterior limbs of internal capsule (n=7), external capsule (n=4), the descending pyramidal tracts (pons in 1, midbrain in 3), and the cerebellar white matter (n=1). A tigroid pattern was found in the centru m se miovale in 8 cases. Nine cases appeared low signal intensity on T_2-weighted i mag es in the thalami; There were 5 cases with mild ventriculomegaly and 1 case with diffuse brain atrophy. Conclusion The typical MR feature of M LD is a bilatera l and symmetrical high signal intensity on T_2-weighted images in periventricu lar and centrum semiovale white matter with initial sparing of the subcortical U -fi ber, and both the genu and splenium of corpus callosum involvement. Besides, the tigroid pattern in the centrum semiovale and involvement of the internal capsul e, external capsule, and corticospinal tract are frequent additional features.

In order to assess whether gene transfection could be mediated by ultrasound in association with P85 and find the appropriate parameters of ultrasound irradiation, the effects of ultrasound with or without P85 on gene transfection of HepG2 cells were examined. The HepG2 cells were irradiated by ultrasound at 1 MHz, 0.4-2.0 W/cm(2) and 50% duty cycle with plasmid encoding enhanced green fluorescent protein (EGFP) as a report gene. Forty-eight h later, the expression of EGFP was detected under the fluorescence microscopy. Transfection efficacy was quantitatively assessed by flow cytometry, and cell viability was evaluated by trypan blue exclusion. The results showed that the transfection efficacy was increased with the increases in ultrasound output power and the ideal transfection efficacy was achieved in HepG2 cells irradiated by ultrasound at 0.8 W/cm(2) for 30 s. The transfection efficacy in ulstrasound+P85 group was three times higher than in single ultrasound group [(17.63+/-1.07)% vs (5.57+/-0.56)%, P<0.05]. The cell viability was about 81% and 62% in ultrasound group and ultrasound+P85 group respectively. It was concluded that ultrasound in combination with P85 could mediate the gene transfection of HepG2 cells, ideal transfection efficacy was achieved by ultrasound irradiation at 0.8 W/cm(2) for 30 s, and P85 could somewhat increase the damage to cells caused by ultrasound.
Cell Survival/genetics ; Green Fluorescent Proteins/genetics ; Hep G2 Cells ; Poloxalene/*pharmacology ; Transfection ; Ultrasonics

Objective To explore the role of inositol requiring enzyme 1 α (IRE1α) mediated endoplasmic reticulum stress associated apoptotic molecules in hippocampal neuronal injury in rats with status epilepsy following lithium-pilocarpine.Methods All 96 Wistar rats were randomly divided into control group and status epilepsy (SE) group.The SE group was further divided into 5 subgroups (3,6,12,24,48 h) according to different time points.pmmunofluorescence was used to observe the expressions of endoplasmic reticulum stress (ERS) markers glucose-regulating protein 78 kd (GRP78) and phosphoIRE1α (active form of endoplasmic reticulum resident protein IRE1α) at the CA3 area of rats in each group.Then,the expressions of IRElα mediated downstream apoptotie markers phospho-c-JunN-terminalkinase (JNK) and caspase12 were detected.Finally,TUNEL assay was used to observe neuronal apoptosis of hippocampal CA3 area at different time points after SE in rats.Results Immunofluorescence showed that GRP78 and phospho-IRE1α positive neurons were significantly increased in the SE subgroups compared with control group (6.90％ ± 0.96％,4.60％ ± 1.12％,respectively) and 12 h subgroup reached the peak (GRP78:87.45％ ±3.63％,F =356.82,P ＜0.05; phospho-IRE1α:86.90％ ±3.82％,F =300.80,P ＜ 0.05).Immunohistochemistry and Western blot demonstrated that the levels of phospho-JNK and caspase12 in the SE subgroups were significantly higher than that in the control group which reached the peak at 12 h after SE.The changes were in accord with phospho-IRE1α.Simultaneously,hippocampal neuronal apoptosis was detected in each SE subgroup and was most severe at 12 h after SE,which showed similar changes to the expressions of phospho-IRE1α,phospho-JNK and caspase12.Conclusions ERS was induced in rats following SE evidenced by increasing the expression of GRP78.IRE1α may promote hippocampal neuron apoptosis in rats following SE through activating JNK and caspase12.

Objective:To investigate the relationship between the level of Bcl 2 protein in EB virus infected cells and the sensitivity of this cells to NK activity and apoptosis inducing factors.Methods:Antisense oligodexynucleotides(ODNs) were used to modulate the expression of Bcl 2 gene in Epstein Barr virus transformed B lymphoblastoid cell lines(EBV LCLs);then the change of cytotoxicity of natural killer cells and the sensitivity of apoptosis inducing elements(taking out growth factor ?dexamethasone)targeting EBV LCLs,were investigated.Results:The level of Bcl 2 expression in EBV LCLs has negative relativity with the cytotoxicity of NK cells to EBV LCLs(P

With the improvement in new drug clinical trials, our hospital has participated in more and more international multi-center clinical trials. Meanwhile, problems arise in carrying out these trials.We here analyzed these problems and put forward some measures for improvement.

Objective To explore the effects of P85,microbubbles and ultrasound on plasmid DNA skeletal muscle gene transduction of mice in vivo. Methods Plasmid encoding green fluorescent protein (GFP) ,which conjugated with 0.05% P85 and/or microbubbles, 10% Optison,was injected into the tibialis anterior(TA) muscle of mice with or without ultrasound irradiation (1 MHz, 1 W/cm2 2 min,20% duty cycle). Mice were killed 1 week after injection. The TA muscles were removed and snap-frozen immediately in isopentane cooled by liquid nitrogen and sections 7 μm thick were cut at intervals. One set of sections mounted with DAPI were used to assess the transfection efficiency by counting the number of GFP-positive fibers under fluorescence microscopy,and the other set of sections were stained with haematoxylin and eosin to assess the tissue damage area. Results The P85 and Optison significantly enhanced the plasmid DNA skeletal muscle gene delivery in vivo separately (P<0.01, P<0.05).Ultrasound exposure could significantly enhance the efficiency of P85 induced gene delivery(P<0.01) but not of Option(P>0.05).The gene delivery efficiency induced by P85 was higher than that by Optison no matter with or without ultrasound irradiation(P<0.01). When the P85 conjugated with Optison, they could further significantly enhance gene delivery efficiency with ultrasound exposure (P<0.01). Meanwhile, ultrasound exposure could increase the muscle damage areas in the groups with microbubbles (P<0.01). Conclusions The P85,microbubbles and ultrasound exposure display synergistic effect to enhance plasmid DNA transduction in skeletal muscle of mice in vivo.

To examine the role of ultrasound in gene delivery in vitro, three cells lines were exposed to the low-frequency ultrasound of varying intensities and for different durations to evaluate their effect on gene transfection and cell viability of the cells. Microbubble (MB), Optison (10%), was also used to observe the role of the microbubbles in gene transfection. The results demonstrated that as the ultrasound intensity and the exposure time increased, the gene transfer rate increased and the cell viability decreased, but at high energy intensities, the cell viability decreased dramatically, which caused the transfer rate to decrease. The most efficient ultrasound intensity for inducing gene transfer was 1 W/cm(2) with duration being 20 s. At the same energy intensity, higher ultrasound intensity could achieve maximal gene transfer rate earlier. Microbubbles could increase ultrasound-induced cell gene transfer rate by about 2 to 3 times mainly at lower energy intensities. Moreover, microbubbles could raise the maximum gene transfer rate mediated by ultrasound. It is concluded that the low-frequency ultrasound can induce cell gene transfer and the cell gene transfer rate and viability are correlated with not only the ultrasound energy intensity but also the ultrasound intensity, the higher ultrasound intensity achieves its maximal transfer rate more quickly and the ultrasound intensity that can induce optimal gene transfer is 1 W/cm(2) with duration being 20 s, and microbubbles can significantly increase the maximal gene transfer rate in vitro.
3T3 Cells ; CHO Cells ; Cell Line ; Cell Survival/*genetics ; Contrast Media/metabolism ; Cricetinae ; Cricetulus ; Microbubbles ; Transfection/*methods ; Ultrasonics

Objective:To study the analgesic effect of preprcelecoxib on total knee arthroplasty in patients with Kashin-Beck disease(KBD).Methods:Eighty-four patients with severe knee joint KBD who underwent total knee arthroplasty in our hospital from January 2017 to January 2019 were selected as the subject of study. The patients were randomly divided into control group ( n = 42) and observation group ( n = 42). The control group was not treated with any analgesia before operation. The observation group was given preemptive analgesia with preprcelecoxib, and patients were injected intravenously with 40 mg preprcelecoxib 30 min before anesthesia induction. The visual analogue score (VAS) score and number of pressing patient-controlled induce analgesia (PCIA) 30 min, 2 h, 4 h and 12 h after operation were compared between two groups. The Hospital for Special Surgery (HSS) score and stress response index [norepinephrine (NE), blood glucose (BG) and cortisol(Cor)] were compared between two groups before operation and 12 and 24 h after operation. And the incidence of adverse drug incidence was also compared between the two groups. Results:The VAS score of the two groups increased at first and then decreased. There was no significant difference in VAS scores between the two groups 30 min after operation ( P > 0.05). But 2, 4 and 12 h after operation, the VAS score in the observation group [(5.75 ± 0.27), (4.02 ± 0.85), (2.04 ± 0.23) points] were significantly lower than those in the control group [(7.34 ± 0.35), (5.96 ± 0.90), (3.89 ± 0.27) points, P < 0.05]. The HSS score increased over time between the two groups, but without significant difference before operation ( P > 0.05). And 12 and 24 h after operation, the HSS scores in the observation group [(53.19 ± 6.53), (75.18 ± 4.63) points] were significantly higher than those in the control group [(46.29 ± 6.37), (60.38 ± 5.29) points, P < 0.05]. The level of stress response index of the two groups increased with time. And 12 and 24 h after operation, the levels of NE [(325.94 ± 25.67), (397.63 ± 27.55) ng/L], BG [(5.38 ± 0.52), (5.41 ± 0.54) nmol/L] and Cor [(241.94 ± 14.18), (253.82 ± 14.65) mg/L] in the observation group were significantly lower than those in the control group [(387.28 ± 26.92), (437.84 ± 28.96) ng/L, (5.79 ± 0.43), (6.54 ± 0.52) nmol/L, (275.39 ± 13.72), (289.63 ± 13.95) mg/L, P < 0.05]. The number of PCIA pressing 24 h after operation in both groups was significantly higher than that 12 h after operation. The number of PCIA (4.62 ± 0.84, 8.38 ± 0.41) in the observation group was significantly lower than that in the control group (8.26 ± 0.65, 12.48 ± 0.73) 12 and 24 h after operation ( P < 0.05). The incidence of adverse reactions (19.05%, 8/42) in the observation group was significantly lower than that in the control group (52.38%, 22/42, P < 0.05). Conclusions:Preprcelecoxib can significantly reduce postoperative pain, reduce the expression of stress index, improve knee motion in a short period of time, reduce the number of pressing (PCIA) and the incidence of adverse drug reactions in patients with severe KBD of knee joint.

Objective To investigate the relationship of gene transfection efficiency with different ultrasound exposure time and different dose of microhuhble,and to find the appropriate ultrasound parameters for gene transfection. Methods Plasmid encoding enhanced green fluorescent protein(pEGFP) was chosen as a report gene and HepG2 cells were chosen as research object. The HepG2 cells plus pEGFP and different dose of microbubble were exposed to ultrasound(1 MHz,0.5 W/cm2) with varying time. Twenty-four hours later, the expression of EGFP in the cells was observed by fluorescence microscope,the transfection efficiency was assessed by FACS and the cell viability was observed by trypan blue exclusion. Results The expression of EGFP in all experimental groups was different,and the approving transfection efficiency was got by ultrasound exposed for 20 s when the dose of microbubble was 60 μl. Conclusions With fixed ultrasound frequency and power, different transfection efficiency was got when the exposure time and dose of microbubble were different. The appropriate parameter was 20 s,60 μl, which can supply information for further study.

Objective To investigate the application value of ELISA for detecting the serum anti desmoglein (Dsg) 1 and Dsg 3 in the diagnosis and treatment of pemphigus .Methods Forty‐seven patients with pemphigus in our hospital from January to De‐cember 2014 were selected as the observation group and contemporaneous 52 patients with excluding pemphigus were selected as the control group .The Dsg antibodies were detected by using indirect immunofluorescence method and Dsg 1 and Dsg3 were deter‐mined by ELISA ;their correlation with pemphigus characteristics was analyzed .Results The sensitivity and specificity of ELISA for detecting anti‐Dsg antibodies were 95 .74% and 92 .31% respectively ,while which of IIF were 93 .62% and 86 .54% respective‐ly ,showing no statistically significant difference between the two test methods (P>0 .05) .In 30 cases of pemphigus vulgaris ,16 ca‐ses (16/30) were positive Dsg1 and Dsg 3 ,8 cases of pemphigus erythematosus and 5 cases pemphigus foliaceus were positive Dsg1 only ,and 2 cases of pemphigus vegetans were both positive Dsgl and Dsg3 .The Dsgl and Dsg3 titers of pemphigus vulgaris and pemphigus vegetans were 130 .85 ± 86 and 112 .30 ± 85 .05 ,respectively ,and the disease activity score was (5 .10 ± 1 .86) points ,the correlation coefficient(r)=0 .476(P=0 .008) ,r=0 .816(P=0 .001) ,respectively .The Dsgl titer of pemphigus erythematosus and pemphigus foliaceus were 142 .59 ± 78 .52 ,and the disease activity score was (2 .77 ± 0 .92) points(r=0 .800 ,P=0 .001) .Conclu‐sion ELISA for detecting Dsg1 and Dsg3 has high sensitivity and specificity ,and is conducive to the diagnosis of pemphigus and e‐valuation of disease severity .