Objective To observe a recombinant mutant of HBV core protein for dominant negative gene therapy against HBV encapsidation in vitro. Methods C gene and S gene of HBV were acquired through PCR and subcloned into pGEM T to construct pGEM T C and pGEM T S respectively. After digestion and ligation of these two plasmids, pGEM T CS was constructed. The cloned gene was inserted into pcDNA3.1 + to construct pcDNA3.1 + CS, which was identified by DNA sequencing. The recombinant plasmids were transformed into HepG2 cells, and screened with G418. The resistant HepG2 cell clones were chosen to test the expression of core surface protein by RT PCR, and the expressing HepG2 clones were cultured with 10% HBV DNA positive human serum for 72 hours. The intracellular HBV particles were extracted and the DNA was subjected to dot hybridization. Results The analysis showed that the HepG2 cells expressing mutant C protein had capabilities to resist HBV invasion in varied degrees. The mutant C protein had a dominant negative role in the encapsidation of HBV compared with the naive part of core protein. Conclusions The production of recombinant mutant core protein has a potential value for gene therapy against HBV infection.

Objective To investigate the relative analysis of prognostic factors in primary anorectal malignant melanoma (AMM). Method Twenty-seven patients with AMM were treated, Log-rank and COX analysis were used to find out the relationship of ten factors to survival. Results Log-rank analysis found five of ten factors affecting survival, they were age of patient,diameter of tumor, infiltrated depth,pathological staging and metastasis (P<0.05 or<0.01), while COX analysis only found the diameter of tumor and infiltrated depth had close relationship to survival (P<0.05 or<0.01). Conclusion The diameter of tumor and infiltrated depth are the main factors affecting the survival of AMM, and this indicates the importance of early diagnosis and treatment.

Objective To measure the macular function of the fellow eye in patients with unilateral retinal vein occlusion (RVO).Methods A total of 24 cases of unilateral RVO were diagnosed by fundus fluorescein angiography (FFA),and multifocal ERG (mfERG) was recorded by RETI scan.The mfERG data of 24 fellow eyes of those RVO patients,and 18 normal control eyes were analyzed and compared.The parameters included the amplitude density,latency of the P1 and N1 wave in 6 concentric circles and 4quadrants of the mfERG graphics.Results The amplitude densities of P1 and N1 wave in first and second concentric circles of RVO fellow eyes were significantly lower than normal eyes (t=4.520,2.147;P＜0.05).There was no significant difference (P＞0.05) of P1/N1 latency in any concentric circles or quadrants between RVO fellow eyes and normal eyes.Conclusion The central fovea of the RVO fellow eyes was functionally impaired.

ObjectiveTo evaluate the application of quantitative sensory testing(QST) in hemodialysis (HD) patients with peripheral neuropathy.MethodsThe values of the quantitative sensory testing and sensory conduction velocity at different locations of extremities in 40 cases of the HD patients and 40 normal persons were recorded.ResultsAccording to the control group,without symptoms group and with symptoms group,the thermal threshold(TT) and vibratory threshold (VT) at the four different locations of extremities were increased,thresholds of the later two groups were significantly higher than those of normal controls (P ＜ 0.05 ).Especially finger warm and vibratory thresholds,hypothenar heat pain thresholds,instep heat pain and vibratory thresholds,calf cold pain and vibratory thresholds were significantly difference between with symptoms group and normal control group (P＜0.01 ).Finger cool and warm thresholds,instep heat pain thresholds,thresholds of calf warm,cold pain and heat pain,and vibratory thresholds at the four different locations of extremities were significantly difference between wthout symptoms group and with symptoms group (P＜ 0.05).The SCV values of the median nerve,tibial nerve and lateral popliteal nerve were significantly difference between HD patients group and normal control(P ＜0.05 ).The total abnormal rate of QST was 65％,with symptoms group was 77.27％,total abnormal rate of SCV was 55.0％,with symptoms group was 68.18％.The total abnormal rate of QST were significantly higher than those of SCV (P＜0.01 ).ConclusionQST is a sensitive method for diagnosis of uremic neuropathy.

Objective To study the relationship between mental effort and personal time perspective during the process of individual changing bad habits.Methods 230 graduate students were collected to attend this longitudinal study of 21 days by experience sampling method.The data on mental effort and performance were obtained from the process of daily change of the bad behavior.Zimbardo Time Perspective Inventory (ZTPI) was applied to graduate students.The survey results were analyzed by hierarchical linear modeling (HLM).Results In 21 days,the mental effort of subjects showed very clear downward trend,the trend of the change was statistical significant (total score(5.63±2.62),t=-5.590,P＜0.01).Mental effort didn't have significant difference in five kinds of behavioral objectives (keeping early hours,exercising daily,dieting,reading daily,limiting screen time) (P＞0.05).The male obviously put more effort than the female during the study(t=2.743,P＜0.01).Both Present Hedonism and Futurism were inversely related to mental effort,which was statistical significant (t=-2.504,P＜0.05;t=2.39,P＜0.05).Both Present Fatalism and Futurism had the effects on the changing trend of the mental effort regulatory,which was statistical significant (t=-3.099,P＜0.01;t=-2.400,P＜0.05).Conclusion The mental effort of changing the behavior will gradually decline during the process of habit establishment.The mental effort of subjects who had present fatalism or futurism show a faster decline.

BACKGROUND: Previous studies have confirmed that atrial natriuretic peptide (ANP) exists in different regions of rat gastric mucosa in different densities, and ANP can inhibit the spontaneous contraction of gastric smooth muscles in rat, guinea pig and human.OBJECTIVE: To investigate the ultrastructural localization of ANP-synthesizing cells and the correlation between ANP-synthesizing cells and microvessel density in rat gastric mucosa.DESIGN: Single sampling study.SETTING: Affiliated Hospital of Chengde Medical College. MATERIALS: Eighteen adult male Wistar rats of clean grade, weighing 300-350 g, were used in this study.METHODS: This study was conducted in the Laboratory of Chinese Materia Medica Immunology, Chengde Medical College from October 2004 to July 2007. The ultrastructural localization of ANP-synthesizing cells in rat gastric mucosa was performed by postembedding immunoelectron microscopy technique. The area density of ANP-synthesizing cell distribution was calculated by histochemical technique (saffron solution and toluidine solution staining included). Microvessels of rat gastric mucosa were revealed distinctively by tannic acid-ferric chloride staining method and scanning electron microscope technique.MAIN OUTCOME MEASURES:①The ultrastructural localization of ANP-synthesizing cells in rat gastric mucosa.②The distribution characteristics of gastric microvessels in different regions of rat gastric mucosa.③The distribution characteristics of ANP-synthesizing cells in rat gastric mucosa.④Correlation between ANP-synthesizing cell distribution and microvessel density.RESULTS: ① Ultrastructural localization of ANP-synthesizing cells: It was confirmed that ANP-cells were the enterochrochromaffin (EC) cells. ② Observation of microvessels: The microvessels were stained successfully by tannic acid-ferric chloride and scanning electron microscope technology. Microvessels of gastric mucous were in wriggled way and cut in different cross-sections. They could be clearly observed in distinct three-dimensions. The microvessel density was the largest in the basal glands of rat gastric mucous. ③ Distribution of ANP-synthesizing cells: It was identified that EC cell was only a chromaffin cell in the rat gastric mucosa. The chromaffin granules (brown granules) were localized in the cytoplasm of EC cells. Negative staining for chromaffin granules was detected in the submucosa and smooth muscle. EC cells had different distribution densities in different regions. The density order of EC cells was gastric cardiac region ＞ gastric pyloric region ＞ gastric fundic region in mucosal layer. ④ Correlation between ANP-synthesizing cells and microvessel density: Correlation analysis showed that there was a positive correlation between ANP-synthesizing cells and microvessel density in the gastric cardiac region(r = 0.53, P ＜ 0.05, n=18), and there was a negative correlation between above-mentioned two indices in the gastric pyloric region(r = 0.38, P ＞ 0.05 , n=18)and in the gastric fundic region(r = -0.29, P ＞ 0.05, n=18).CONCLUSION: ANP-synthesizing cells are the EC cells of gastric mucosa. There is a positive correlation between ANP-synthesizing cells and microvessels in the rat gastric cardiac region, while there is a negative correlation between ANP-synthesizing cells and microvessels in the rat gastric pyloric region and in the gastric fundic region.

Background The severity of difficulty in falling asleep and the insufficiency of the sleeping time in the primary insomniacs always were overestimated, even after medication and improving the objective sleep quality. Moreover, remarkable psychosomatic symptoms were found in patients and the symptoms were significantly linked to the self-reported sleep quality. So in this study, it was designed to explore the differences between objective and subjective sleep quality and its correlative factors in chronic insomniacs.Methods A total of 55 chronic primary insomniac cases diagnosed by DSM-Ⅳ diagnostic criteria for primary insomnia and 15 normal controls were rated with Pittsburgh sleep quality index (PSQI),State-Trait Anxiety Inventory (STAI), and the Subjective General Condition Form. All cases and the controls were then examined by the whole-night polysomnography (PSG) records.Results Compared with the objective parameters of PSG, the subjective parametersiu chronic insomniacs had significantly longer sleep latency and lower total sleep time and sleep efficiency [ (80. 3± 73.7)min v.s. (23.2 ± 25.4) min, (157.8±141.7)min v.s. (332.2±154.7)min , (0.52 ±0. 27) min v.s. (0. 67 ± 0. 28) min, respectively, P ＜0. 001 ]. The difference between subjective and objective sleep latency was related to the total scores of STAI, TAI and SAI ( r = 0. 402,0. 374 and 0. 397, respectively, P ＜ 0. 05).Conclusions There were significant differences between objective and subjective estimations of sleep quality in chronic insomniacs,and they might result in the overestimation of the severity of insomnia. The differences between subjective and objective sleep quality in the chronic insomniacs were significantly correlated with the levels of anxiety.

A new cool-tip radiofrequency (RF) ablation therapeutic instrument based on impedance control algorithm is introduced in this paper. The equipment is composed of hardware system and software system. The RF power output and real time data acquisition are completed by the hardware system, while the software is used mainly to finish the control of the ablation range, the core of which is impedance control algorithm, and it also used to complete the display of the real time data in the course of the experiment. The impedance algorithm has solved the problem of impedance increased rapidly during the RF ablation, which has also expanded the scope of ablation. The pig liver experiments showed that the impedance control algorithm had strong adaptability. It also obtained a result of ablation range up to 3.5-4.5 cm single needle. It has the high clinical practical value of one-time inactivation of 3-5 cm tumor.
Algorithms ; Animals ; Catheter Ablation ; Electric Impedance ; Humans ; Liver ; Needles ; Software ; Swine

Objective To investigate the changed brain areas of functional connectivity ( FC ) patterns of subthalamic nucleus ( STN) between different subtypes of Parkinson′s disease ( PD) and healthy controls and their clinical significance .Methods Twenty-three PD patients and 12 health controls were enrolled, and PD patients were divided into 11 tremor dominant PD patients and 12 postural instability and gait disorder (PIGD) dominant PD patients.The difference of whole-brain resting-state FC with STN among the three groups was investigated and the relationship between the changed areas and clinical symptoms was calculated.Results Compared with healthy controls , PIGD dominant group showed increased FC between STN and right calcarine and decreased FC between STN and right precuneus , and tremor dominant PD patients showed decreased FC between STN and precuneus , right angular gyrus and left middle frontal gyrus , while the FC strength between the STN and left middle frontal gyrus showed positive relationship with MMSE scores ( r=0.64, P=0.034 ) .Compared with tremor dominant PD patients , PIGD dominant patients showed increased FC in the area of left calcarine .Conclusions Decreased FC between STN and precuneus was gained in different subtypes of PD , and PIGD dominant patients showed increased FC between STN and calcarine.Changed areas based on the FC of STN in different subtypes of PD were found related with cognition and mood control .

Objectives To investigate MRI patterns of functional connectivity(FC)in different brain areas of the subthalamic nucleus (STN)in Parkinson's Disease (PD)and its correlation with cognition.Methods We used functional magnetic resonance imaging to investigate the difference in whole-brain resting-state FC of STN between 32 patients with PD during the medicatiom ON state and 25 healthy control group(HC)matched for age,gender,and cognition,and examine the correlation between functional connectivity strength and montreal cognitive assessment (MoCA)scores.Results Compared with HC,the PD group showed increased FC in the right lingual gyrus of the left STN and the right STN showed decreased FC in the left superior frontal gyrus and the supplementary motor area(t=4.29,-3.61,and-3.83,respectively,each P ＜ 0.05),while the right STN showed only decreased FC in the right bilateral cingulate gyrus and the precuneus(t=-4.44,4.29,and-4.30,respectively,each P＜ 0.05).In addition,PD patients' connectivity strength between RSTN and the bilateral precuneus was positively correlated with MoCA scores(t =0.58 and 0.57,respectively,each P＜0.05).Conclusions Compared with HC,PD patients exhibit decreased FC between RSTN and the precuneus,with FC strength positively correlated with MOCA scores.The cognitive decline caused by deep brain stimulation in STN may be related to injuries of the precuneus.