Objective Getting the robust exogenous gene expression vector under the control of porcine insulin promoter, and to lay the foundation for pancreaticβ-cells specific transgene expressing pigs.Method Using porcine insu-lin promoter ( PIP, 1500 bp of the 5′UTR from the porcine INS gene including the first exon and the first intron) to con-struct expression vector, the HindIII restriction site which connected the sequences of PIP and EGFP was designed before ATG, named PIP-HindIII-EGFP.Considering that the different location of restriction site may affect the expression efficien-cy of the transgene, we optimized the expression vector.Firstly the HindIII restriction site was deleted to realize the seam-less connection of PIP and EGFP,the vector was named PIP-EGFP.Also we mutated the 3′intron splicing acceptor site( SA) of the first intron into HindIII restriction site, named as PIP-SA( M)-EGFP.Three different EGFP expression vectors were respectively transfected MIN-6 mouse pancreatic β-cells, pig ear fibroblasts and kidney cells.The transfected cells were cultured for 48 h and harvested for RT-PCR, flow cytometry and Western blot analysis, to analyze and compare the expres-sion efficiency of vectors.Results After transfection,green fluorescence was observed only in MIN-6 mouse pancreaticβ-cells.RT-PCR analysis and product sequencing showed that the three expression vectors did have different stability with in-tron splicing.The PIP-HindIII-EGFP construct and PIP-EGFP vector produced two kinds of mRNA with the first intron spliced and no spliced, indicating the instability of intron splicing.Mutation of the PIP splice site would cause the first in-tron not spliced, while flow cytometry and Western blot displayed that the mutation induced a most efficient expression of the downstream gene.Conclusions A robust and specific β-cells expression vector has been successfully generated by mutating the intron splicing acceptor site of the porcine insulin promoter.It provides the foundation for preparation of pigs with pancreaticβ-cells specifically expressing the transgene.

Objective To explore mechanisms of sustained and transient effects in encoding processes of emotional memory by examining activation of amygdala via functional MRI and to provide evidence for understanding the underlying neural mechanism related to emotional memory disorders further. Methods Twenty two subjects (aged from 20 to 24 years old) participated in the study and mixed blocked/event-related design was adopted. Sixty negatively emotional pictures and sixty neutral scene pictures were used. Functional MRI scanning was performed while subjects were doing encoding tasks. Behavioral data were acquired during retrieval. Correlation analyses of functional MRI data and simples paired t -test of behavioral performance were performed with SPM2 and SPSS13.0 statistical software,respectively. Results Significant differences of behavioral performance ( t= 2.791,P= 0.01 ) was found between emotional (3.15 ± 0.14) and neutral (2.25 ± 0.08 ) pictures. A whole-brain voxelwise correlation analysis between functional MRI and emotional enhancement effect indicated that the transient effect of emotional enhancement of memory involved the left amygdala, left hippocampus and left lateral orbitofrontal cortex, while the sustained effect involved the right amygdala, right hippocampus, right inferior frontal gyrus, right medial and lateral orbitofrontal cortex. Region of interest analysis demonstrated that the sustained effect was related to the right amygdala (r= 0.50, P = 0.019 ), which was different from transient effect ( Z = 1.655, P = 0.049 ),while the transient effect was correlated with the left amygdala (r=0.65, P=0.001 ) ,which was different from sustained effect( Z= 2.512, P=0.006). Conclusion Different neural mechanisms are involved in sustained and transient effects of emotional memory encoding. The right amygdala is responsible for sustained effect and the left amygdala is responsible for transient effect ,respectively. The results confirm and extend the model of the left-transient/right-sustained effect.

Objective To approach the protective effect of dexmedetomidine on mechanically ventilated patients with pulmonary contusion. Methods A prospective randomly controlled trial was conducted. 70 mechanically ventilated patients with pulmonary contusion from January 2010 to December 2012 in the Critical Care Medicine of Foshan Hospital of Traditional Chinese Medicine were divided into a control group and a therapy group by the difference in number odd or even,with 35 patients in each group. Based on the same principles of comprehensive treatment,the control group used midazolam,and the therapy group used dexmedetomidine for sedation. The measured parameters included oxygenation index(PaO2/FiO2),tumor necrosis factor-α(TNF-α),interleukin-6(IL-6), and extra-vacular lung water index(EVLWI)for both groups on the1st and 5th day. The incidence of delirium,the time of mechanical ventilation,and the incidence of hypotension were observed in both groups. Results Compared with those on the 1st day,TNF-α,IL-6 and EVLWI on the 5th day were decreased significantly in both groups〔the control group TNF-α(ng/L):1.29±0.38 vs. 2.21±0.37,IL-6(ng/L):97.97±28.77 vs. 131.03±41.52,EVLWI (mL/kg):8.25±2.03 vs. 11.96±3.36;the therapy group TNF-α:1.06±0.33 vs. 2.32±0.37,IL-6:82.07±23.35 vs. 134.98±64.25, EVLWI(mL/kg):6.74±1.33 vs. 11.23±2.78, all P<0.05〕, PaO2/FiO2 was increased obviously〔mmHg(1 mmHg=0.133 kPa),the control group:285.80±30.65 vs. 213.00±33.70,the therapy group:315.00±34.50 vs. 229.50±37.00,both P<0.05〕,TNF-αand IL-6 had no significant difference compared between the therapy group and control group(TNF-α:1.06±0.33 vs. 1.29±0.38,IL-6:82.07±23.35 vs. 97.97±28.77), while EVLWI and PaO2/FiO2 in the therapy group had remarkable differences compared with those of the control group〔EVLWI(mL/kg):6.74±1.33 vs. 8.25±2.03,PaO2/FiO2(mmHg):315.00±34.50 vs. 285.80±30.65,both P<0.05〕. The incidence of delirium(8.57% vs. 22.86%)and time of mechanical ventilation(day:4.10±1.09 vs. 6.88±1.66)in the therapy group were decreased markedly compared with those of the control group,and the incidence of hypotension had no significant difference between treatment or control groups(17.14% vs. 14.29%,P>0.05). Conclusion Dexmedetomidine has protective effect on mechanically ventilated patients with pulmonary contusion, and it is an relatively ideal sedative drug for these patients.

Objective:To investigate the expression of IGF-IR and PKC and their clinical significance in laryngeal squamous cell carcinoma(LSCC).Method:The expression of IGF-IR and PKC protein were detected by immunohistochemistry and Western blot in 60 cases of LSCC, 25 cases of adjacent non-tumorous laryngeal epithelium and 10 cases of normal laryngeal epithelium.Result:The positive rate of IGF-IR and PKC expression in LSCC were significantly higher than that in normal laryngeal epithelium and adjacent non-tumorous laryngeal epithelium(P<0.05). IGF-IR positive expression were increased more in cases with stage Ⅲ-Ⅳ, cervical lymph node metastasis and well differentiation than that in cases with stageⅠ-Ⅱ, non-lymph node metastasis and poor/mediate differentiation, respectively(P<0.05).The positive rate of PKC in LSCC were correlated with clinical stage and cervical lymph node metastasis.There was a correlation between the expression of IGF-IR and PKC in LSCC.Conclusion:The overexpression of IGF-IR and PKC in the laryngeal carcinoma may play an important role in the carcinogenesis and development of LSCC. It is suggested that detecting the expression of IGF-IR and PKC can be used for early diagnosis and treatment of LSCC.

Objective To evaluate the clinical value of fiberoptic ductoscopy in diagnosis and treatment of patients with galactophoritis or mammary duct ectasia. Methods From November 2005 to March 2008, fiberoptic ductoscopy were performed in 120 women with nipple discharge. The duct of 95 cases as non-malignant lesion were insufflated and perfusioned with entamycin and dexamethasone. Results Ninty-five of 120 cases were non-malignant disease,which contained one side 81 and two sides 14; the discharge was bloody,ivory, stramineous in 21, 17, 57 patinents, respectively; and the dignosis were 17 mammary duct ectasia, 53 galactophoritis, and 25 mammary duct ectasia with galactophoritis. Of the 95 cases, hich were intradutal insufflated and perfusioned with gentamycin and dexamethasone, the nipple discharge were decreased or disappeared in 81 cases, the effective rate was 85.3%. Conclusion Fiberoptic ductoscopy is a convenient,safe, accurate method in diagnosis and treatment of patients with galactophoritis or mammary duct ectasia.

Objective To establish a cosmetic medicine bio-psycho-social model and its theory frame and a new clinical work system. Methods The clinical observation and clustering analysis were used in this study, which based on the modern medicine patterns to carry on the logical proof, theory derivation and systematic design. Results The bio-psycho-social model of cosmetic me dicine with combination of the theory and practice was established, including its concept, theory constitution and clinical work system. Conclusion The bio-psycho-social model of cosmetic medicine can most scientifically and comprehensively explain this discipline rule and clinical work, and it will bring the profound influence on the development of the theory and practice in cosmetic medicine.

0.05).Conclusion: Enteral nutrition may improve the nutrition status.The naso-jejunum tube feeding has no advantage to nasogastric tube feeding in our patients.The route of enteral nutrition should be selected according to the individual condition and local nosocomial experience.

objective To observe the distribution of vitamin D receptor(VDR)gene polymorphisms in children of Han nationality and investigate the relationship between VDR gene polymorphisms and the susceptibility to children's dental fluorosis of Han nationality.Methods From October of 2008 to March of 2009,a case-control study was conducted among children between 8 and 12 years old with(n=101)and without(n=102)dental fluorosis using Dean method in Guandian countyside of Fengtai county in Anhui province.DNA was extracted from blood samples ofthese children.The Apa I,Bsm I,Fok I and raq I polymorphisms in the VDR gene were genotyped using polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP).The distribution of the genotypes in patients and the control group were analyzed. Results Different genotypes of the VDR gene existed in children of Han nationality with the highest distribution frequency of Aa, bb, Ff, TT, which respectively was 51.7% ( 105/203 ), 89.7% (182/203), 52.7% (107/203), 93.1% (189/203), followed by genotype distribution frequency of aa, Bb, FF, Tt, being 39.9% (81/203),7.9% (16/203),31.5% (64/203) ,6.9% (14/203), respectively. AA,BB, ff, tt distribution frequency was the lowest as follows, 8.4% ( 17/203 ), 2.4% (5/203), 15.8% (32/203),0 (0/203). The frequency distribution of VDR Apa I genotype was AA 7.9% (8/101), Aa 55.4% (56/101), aa 36.7%(37/101) in children with fluorosis, and AA 8.8% (9/102), Aa 48.0% (49/102), aa 43.3% (44/102) in children without fluorosis, respectively. There were no significant differences in the two groups(χ2= 1.13, P > 0.05).The frequency distribution of VDR Bsm I genotype was BB 3.0%(3/101), Bb 5.9%(6/101 ), bb 91.1% (92/101) in children with fluorosis, and BB 2.0% (2/102), Bb 9.8% (10/102), bb 88.2% (90/102) in children without fluorosis, respectively. There were no significant differences in the two groups(χ2 = 0.55, P > 0.05). The frequency distribution of VDR Fok I genotype was FF 28.7%(29/101), Ff 56.4% (57/101), ff 14.9%(15/101) in children with fluorosis, and FF 34.3% (35/102), Ff 49.0% (50/102), ff 16.7% (17/102) in children without fluorosis,respectively. There were no significant differences in the two groups(χ2 = 1.14, P > 0.05). The frequency distribution of VDR Taq I genotype was TT 93.1%(94/101), Tt 6.9%(7/101) in children with fluorosis, and TT 93.1% (95/102), Tt 6.9%(7/102) in children without fluorosis, respectively. The tt genotype was not found. There were no significant differences in the two groups (χ2 = 0.00, P > 0.05 ). Conclusions Different genotypes of the VDR gene existed in children of Han nationality. There were no correlation between VDR Apa I , Bsm I , Fok I , Taq I gene polymorphisms and children's dental fluorosis of Han nationality in this area.

Objective:It is demostrated that the porous protein-mineral mechanics model could provide more accurate prediction for biomaterial properties of dentine compared with the other established models.This paper would use the model to reevaluate the mechanical properties and its interacting mechanism of human dentine.Method:By using a porous proteinmineral mechanics model,the effect from the interactions between tubules,pedtubular and intertubular matrix on dentine microstructure was discussed.Results:The dentinal micromechanical properties were dependent on the tubular direction,and the absolute values of the stresses derived from the hydraulic and gas tubular pressures increased parabolically with theincreasing diameter of the tubules.It was also found that the effective elastic constants of the dentine microstructure would vary with the aging and the distribution of mineral and collagen within peritubular and intertubular matrix of detine.Conclusions:The theoretical analyses provided in this paper demonstrated that the microstructural characteristics of tubules,peritubular and intertubular dentinal matdx could have different influences on the micromechanical properties of human dentine,which showed the validity of porous protein-mineral mechanics model,and the limitation of some models that neglected the interacting mechanism.

Objective The brain volume and behavioral performance of emotional memory were measured by combining voxel-hased morphometry (VBM) and behavior to explore the morphological characters of brain regions related to the emotional enhancement effect of memory.Methods A total of 32 healthy young subjects participated in the study.The experimental processes included MR scan and behavioral performance of emotional memory the latter of which involved two phases-encoding and retrieval.Behavioral performance was recorded and the whole-brain 3D MRI data were acquired via 3.0T MR.3D MRI data were segmented by VBM5,and the wholebrain volumes of gray matter,white matter and CSF were produced.SPM5 and SPSS13.0 were used for statistical analysis of the 3D MRI data and behavioral data,respectively.Results Retrieval accuracy of the emotional pictures was higher than that of the neutral pictures ( t =5.08,P＜0.001 ),the emotional enhancement effect was significant ( Δ Pr =0.12 ± 0.01 ).Brain regions related to emotional enhancement effect included bilateral amygdala,dorsolateral prefrontal cortex,middle frontal gyrus,dorsomedial prefrontal cortex,fusiform gyms,right parahippocampus,left orbitofrontal cortex and middle cingulate gyms.Volume ratio of bilateral amygdala were related to emotion enhancement effect evidently (left:r=0.564,P＜0.01 ; right:r=0.541,P＜0.01 ).Conclusion This study morphologically confirms that brain regions,such as amygdale and prefrontal cortex,are critical structures for the enhancement of emotional memory,and further extends the findings of previous functional imaging studies.