Objective To study the effect of selective COX-2 inhibitor NS-398 on IL-1α and TNF a expression in HaCaT cells induced by UVA/UVB,and further to explore the mechanism on anti human skin photo damage. Methods The subcuhured HaCaT cells were divided into three groups:simple illuminated group was exposed to UVA/UVB directly,NS-398 interfered group was exposed to UVA/UVB after being treated with NS-398 in different concentrations,and the control group was cultured in normal without any treatment.The expression of IL -1α and TNF-α in supernarant was detected by ELISA kit.Results The level of IL-1α and TNF-α expression in supernatant from simple illuminated group was remarkably higher than that in the control group,NS-398 interfered group showed much lower level than the simple illuminated one,and the expression of IL-1α and TNF-α was dependent on the concentration of NS-398.Conclusions NS-398 can reduce IL-1α and TNF-α expression in HaCaT cells induced by ultraviolet rays,suggesting the possibility of anti human skin photo- damage effect.

Objective To study the effect of selective COX-2 inhibitor NS398 on IL-10 and IFN-γexpression in human keratinocytes induced by UVA,and further to explore the effect on anti human skin photoaging.Methods HaCaT cells were divided into three groups:simple UV exposure group,NS398-intervene group and blank group.For UV radiation,simple UV exposure group and NS398-intervene group were irradiated with UVA at a dose of 30 J/cm2.In order to assess the effect of NS398,HaCat cells were treated with NS398 at the dose of 0,20,40,80μmol/L respectively and then incubated without irradiation for 2h,substrate changed and cultured for 24h,and then the expression of IL-10 and IFN-γ mRNA was detected via RT-PCR.Results Simple UV exposure group had obviously higher expression of IL-10 mRNA and lower expression of IFN-γ mRNA than that of control group,while NS398-intervened group had significantly lower expression of IL-10 mRNA and higher expression of IFN-γ mRNA than that of simple UV exposure group,and dose-dependency existed.Conclusions NS398 may delay photo-damage of human skin via inhibiting the expression of IL-10 mRNA and up-regulating the expression of IFN-γ mRNA in human keractinocytes.

With the development of functional genomics, high throughput analysis of genes’ function has been the mainstream of research, and exogenous gene's over expression via Agrobacterium-mediated transformation is the most commonly used method in gene functional analysis.The versatile plant expression vector cassette named pBHT-5 was constructed by the method of site-specific mutagenesis based on pBI121. First of all, the restriction enzyme SfiI recognition site in trfA gene (X00713) which was relevant to plasmid replication and stability was replaced without changing its amino acid composition. And then the SfiIA,SfiIB sites were added between promoter CaMV35s and terminator NOS. The versatile plant expression vector cassette can be directly used to construct plant expression vector containing the full-length genes cloned by Clontech SMARTTM technology, which will raise the efficiency of vector construction. The result will provide basis of new genes’ high throughput screening and functional analysis, then get the new genes functioning in plant osmotic stress resistance.

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjcl4FABP and Sjc26GST genes and identify their expression in vitro, Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human placental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sj14 or pVAC-Sj26 only to get two gene fragments including Sj14 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES, resulting in another new recombinant plasmid pIRES-Sj26-Sj14. The expression of Sj14 and Sj26 genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and plRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.

Objective To investigate the effect of different duration skeletal muscle denervation on acetylcholine receptor activity in rats.Methods Fourteen Balb/c mice weighing 18-22 g were used in this study.The denervation model was established by excising sciatic nerve.Two rats were chosen before(T0 ) and at days 1,4,7,14,21 and 28 after excising sciatic nerve (T1～6),and flexor digitorum brevis of the hindfoot was acutely isolated Skeletal muscle cells were isolated ( five cells in each rat),the acetylcholine currents were recorded using whole-cell patch-clamo technique.Extracellular fluid containing 30 μmol/L acetylcholine was first applied to skeletal muscle cells for 10 s,acetylcholine currents (11)were recorded,then the ceils were washed out using extracellularfluid.Skeletal muscle cells were balanced using extracellular fluid containing 0,0.1,1,10,30,100,1000,3000,or 10 000 nmol/L atracurium for 3 min respectively,then perfused using extracellular fluid containing 30 μmol/L acetylcholine and differents concentrations of atracurium mentioned above for 10 s respectively,and acetylcholine currents were recorded,then the cells were washed out,and 30 μmol/L acetylcholine was perfused again and currents(I2 ) were recorded.The mean value of I1 and I2 was taken as control current,and inhibitory percentage of control current was calculated,and the inhibition concentrations for the half-maximal response (IC50) of atracurium were determined by nonlinear regression analysis.Results Compared with T0,IC50 significantly increased at T1～6 ( P ＜ 0.05).IC50 was increased gradually at T1～3 ( P ＜ 0.05).Compared with T3,IC50 was decreased at T4～6 ( P ＜ 0.05).IC50 was decreased gradually at T4～6 ( P ＜ 0.05 ).Conclusion Skeletal muscle denervation can inhibite acetylcholine receptor activity,which is relate to the denervation time.

BACKGROUND: Traditional scaphotrapeziotrapezoid limited intercarpal arthrodesis contains Kirschner wire, U-shaped nail, AO/ASIF steel plate and so on. Long-term plaster external fixation was needed following surgery. USA-designed arthrodesis apparatus is mainly suitable for European and people from USA, but not fit for Asian.OBJECTIVE: To simulate scaphotrapeziotrapezoid limited intercarpal arthrodesis, and to test the stability of the novel arthrodesis apparatus developed by the group according to anatomic form of dorsal joint fovea of Chinese scaphotrapeziotrapezoid.DESIGN, TIME AND SETTING: The observational study was performed at the Laboratory of Biomechanics of the Department of Orthopaedics, Nantong University from April 2006 to March 2007.MATERIALS: A total of 40 fresh forearm samples of corpses that were not subjected to antiseptic treatment were used for this study. Radiograph verified that these samples did not develop wrist joint affection or abnormal alinement.METHODS: The cadaver models were imitated limited intercarpal instability before scaphotrapeziotrapezoid arthrodesis used circular plate. Then, simulated flexion 50°, extenion 35°, ulnar deviation 30°, radial deviation 10° ultimate activities (50000).CT scan and 3-D reconstruction should be taken before and after motion.MAIN OUTCOME MEASURES: The index of the radial-scapho angle (RSA), radial-scapho distance (RSD), scapho length(SL), trapezium-trapezoid inclination (TTI), trapezium-trapezoid width (TTW) were measured.RESULTS: Prior to motion RSA, RSD, SL, TTW and TTI were (38.725±2.230) °, (18.988±1.216) mm, (1.686±0.191) cm,(27.360±1.571) mm and (114.975±2.293) °. Following motion RSA, RSD, SL, TTW and TTI were (38.800±2.388) °,(19.215±1.443) mm, (1.683±0.209) cm, (27.718±1.910) mm, (115.300±3.023) °. No significant difference in above-described indexes was determined before and after motion (P > 0.05).CONCLUSION: The novel STT limited intercarpal arthrodesis apparatus shows confidential stability. Thus, wrist joint can do exercises in an early stage at the range of 35 dorsal extension, 50 palmar flexion, 10 radial deviation and 30 ulnar deviation.

Objective: To investigate the prevalence of current smoking and smoking cessation, and identify the influencing factors among hypertensive patients in Nanshan District, Shenzhen City, so as to provide insights into smoking control among hypertensive patients. Methods: The demographic features, life style, status of smoking and smoking cessation in hypertensive patients were collected from 69 community health centers in Nanshan District from 2017 to 2019. The gender- and age-specific prevalence of smoking and smoking cessation was estimated among hypertensive patients, and the factors affecting hypertensive patients' smoking and smoking cessation were identified using a multivariable logistic regression model. Results: Totally 4 385 patients with hypertension were enrolled, with a mean age of (58.41±11.19) years, and the participants included 2 265 men (51.65%) and 2 120 women (48.35%). There were 724 current smokers, and the prevalence of current smoking was 16.51%, with 29.71% prevalence in males and 2.41% in females. There were 424 hypertensive patients quitting smoking, and the prevalence of smoking cessation was 36.93%, with 38.20% prevalence in males. Multivariable logistic regression analysis identified male ( OR=12.885, 95%CI: 9.567-17.354 ) and drinking ( OR=2.567, 95%CI: 2.118-3.111 ) as facilitating factors for current smoking among hypertensive patients, and increasing age (OR=0.723, 95%CI: 0.642-0.815) and high exercise frequency (OR=0.881, 95%CI: 0.817-0.950) as barrier factors, while male ( OR=7.309, 95%CI: 3.304-16.165), increasing age ( OR=1.381, 95%CI: 1.120-1.703 ), unmarried ( OR=1.819, 95%CI: 1.329-2.490 ), divorced ( OR=7.837, 95%CI: 1.254-48.975 ), retired ( OR=1.545, 95%CI: 1.095-2.180 ), unemployed (OR=1.801, 95%CI: 1.057-3.066), and high exercise frequency ( OR=1.221, 95%CI: 1.096-1.360 ) were identified as facilitating factors for smoking cessation among hypertensive patients and widowed ( OR=0.285, 95%CI: 0.089-0.906 ), high educational level ( OR=0.766, 95%CI: 0.619-0.949 ), and drinking ( OR=0.488, 95%CI: 0.368-0.647 ) as barrier factors. Conclusions The prevalence of smoking is lower, and the prevalence of smoking cessation is higher among hypertensive patients than among general populations in Nanshan District. Young and middle-aged, employed, widowed men with a high educational level are key populations for tobacco control, and alcohol consumption control and intensified exercises are important measures to reduce the prevalence of smoking and improve the prevalence of smoking cessation.

OBJECTIVETo observe the expression and clinical implication of receptor for activated C kinase 1 (RACK1) in mononuclear cells and coronary atherosclerotic plaques from patients with coronary artery disease.

METHODSmRNA and protein expressions of RACK1 were detected in mononuclear cells from 29 patients with stable angina pectoris (SAP), 41 patients with acute coronary syndrome (ACS) and 30 healthy volunteers. RACK1 protein expression was also detected by immunohistochemistry in 17 coronary atherosclerotic plaques and 6 normal autopsy coronary samples.

RESULTS(1) mRNA expression of RACK1 was significantly upregulated in mononuclear cells from patients with ACS compared with those from patients with SAP (18.71 ± 5.45 vs. 12.18 ± 4.14, P < 0.05), and the latter was also significantly higher than in healthy controls (12.18 ± 4.14 vs. 3.65 ± 1.57, P < 0.05). (2) Similar changes were observed for protein expression of RACK1 for the three groups. (3) Increased expression of RACK1 was found in atherosclerotic plaques, especially in unstable plaques, positive RACK1 stain was evidenced in foam cells, inflammatory cells, smooth muscle cells and endothelial cells.

CONCLUSIONSThe expression of RACK1 is significantly upregulated in mononuclear cells from patients with coronary artery disease, especially in patients with ACS, and in coronary atherosclerotic plaques, especially in unstable plaques. Our results thus suggest that RACK1 might play an important role in the development and progression of coronary artery disease.

Adult ; Aged ; Case-Control Studies ; Coronary Artery Disease ; blood ; genetics ; pathology ; Female ; Gene Expression ; Humans ; Leukocytes ; metabolism ; Male ; Middle Aged ; Receptors for Activated C Kinase ; Receptors, Cell Surface ; genetics ; metabolism

To further uncover the scientific significance and molecular mechanism of the Chinese herbs with pungent hot or warm natures, endogenous and exogenous expression systems were established by isolation of dorsal root ganglion (DRG) neurons and transfection of HEK293 cells with TRPV1 channel gene separately. On this basis, the regulation action of capsaicin, one main ingredient from chili pepper, on TRPV1 channel was further explored by using confocal microscope. Besides, the three-sites one-unit technique and method were constructed based on the brown adipose tissue (BAT), anal and tail skin temperatures. Then the effect of capsaicin on mouse energy metabolism was evaluated. Both endogenous and exogenous TRPV1 channel could be activated and this action could be specifically blocked by the TRPV1 channel inhibitor capsazepine. Simultaneously, the mice's core body temperature and BAT temperature fall down and then go up, accompanied by the increase of temperature of the mice's tail skin. Promotion of the energy metabolism by activation of TRPV1 channel might be the common way for the pungent-hot (warm) herbs to demonstrate their natures.
Adipose Tissue, Brown ; drug effects ; physiology ; Animals ; Capsaicin ; analogs & derivatives ; pharmacology ; Energy Metabolism ; Ganglia, Spinal ; cytology ; HEK293 Cells ; Humans ; Mice ; Neurons ; drug effects ; physiology ; Plants, Medicinal ; chemistry ; TRPV Cation Channels ; physiology ; Temperature ; Thermogenesis

Objective To investigate the efficacy of micro botulinum toxin A (BTX) combined with intense pulsed light (IPL ) treatment in facial rejuvenation patients .Methods A total of thirty patients (1 male ,29 females ,aged 35 to 55 years with average age of 45 .56 years) with facial pho-toaging in the Department of Medical Cosmetology ,Renmin Hospital of Wuhan University from Jan . to Sept .2018 were enrolled as out-patients .Thirty patients were recruited in the study and randomly assigned averagely to the combination group and control group .All received standard IPL treatment every month for a total of 3 sessions .Patients in combined group received microinjections of diluted BTX on cheeks after the first IPL .VISIA CR images were analyzed before ,1 month and 3 months af-ter treatment .Dermatologists assessed the improvement of color spots ,wrinkles ,texture ,pore ,and erythema improvement .The patient satisfaction survey was analyzed .Results The effective rate of the combined group was better than that of the intense pulsed light group one month after treatment (χ2 = 6 .208 ,P = 0 .045) ,but there was no significant difference between the two groups three months after treatmen (χ2 = 1 .077 ,P = 0 .584t) .The patient satisfaction survey showed that there was no sta-tistical difference between the two groups .Slight erythema and bruising were observed but disappeared at the end of the treatment (P > 0 .05) .Conclusions The adjunctive use of MicroBTX can enhance treatment efficacy of IPL alone .It is a highly effective and safe treatment method for facial antiaging .