AIM:ToexplorewhetherMCP-1-JAK2/STAT3signaltransductioninthespinaldorsalhornin-volves the formation and development of rat type 2 diabetic neuropathic pain (DNP).METHODS: The male Sprague-Dawley rats were fed with a high-fat and fructose diet for 8 weeks,and then received a single intraperitoneal streptozocin in-jection to prepare the type 2 DNP model.The type 2 DNP rats were randomly divided into 4 groups (n=16):DNP group, MCP-1 neutralizing (DM) group, DNP+AG490 (DA) group and solvent control (SC) group.A catheter of PE-10 was placed into the subarachnoid space of the rats in groups DM , DA and SC.After 3 d, the rats in DM,DA and SC groups were injected with MCP-1 inhibitor 10μL at 0.1 mg/L, AG490 10μL at 1 mmol/L and DMSO 10μL at 3.5%once a day for 14 days, respectively.Another 16 normal rats were selected as control (C) group and were fed with common forage. The mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were measured at 1, 3, 7 and 14 d after subarachnoid injection .The lumbar segments 4-6 of the spinal cord were removed at the same time for determination of the expressions of p-JAK2 and p-STAT3 by Western blot .RESULTS:Compared with C group , MWT was significantly de-creased, TWL was shortened and the expression of p-JAK2 and p-STAT3 in the spinal dorsal horn was up-regulated at 1, 3, 7 and 14 d in DNP and SC groups (P<0.05).Compared with DNP group, MWT was significantly increased, TWL was prolonged and the expression of p-JAK2 and p-STAT3 in spinal dorsal horn was down-regulated at 1, 3, 7 and 14 d in DM and DA groups (P<0.05).No significant difference in the MWT, TWL and expression of p-JAK2 and p-STAT3 between DNP group and SC group was observed (P>0.05).CONCLUSION:The MCP-1-JAK2/STAT3 signal transduction in the spinal dorsal horn involves the formation and development of DNP in rats .

10.3969/j.issn.2095-4344.2013.26.015

Objective To investigate the morphology of proximal contact areas (PCA) and gingival embrasures of the maxillary anterior dentition in Shanghai adolescents, hoping to provide evidence for clinical treatment of maxillary anterior tooth esthetic restoration. Methods Totally 62 dental models of Shanghai adolescents with normal occlusion were collected with silicon rubber, and each case was scanned by Sirona inlab inEos Blue and measured by CAD/CAM bundled software. The collected data were analyzed by SPSS18.0 software. Results The mean proximal contact area proportion (PCAP) between central incisors (CI-CI), central and lateral incisors (CI-LI), and lateral incisors and canines (LI-CA) were (47. 7 ± 5. 1) %, (34.6 ± 5.0)%, and (24. 2 ± 4. 2)% in central incisor of square shape, (43. 1 ± 3. 7)%, (31. 3 ± 4. 1)% and (22. 3 ± 3. 7)% in ovoid shape, and (38. 8 ± 5. 3)%, (33. 1 ± 5. 7)% and (23. 2 ± 3. 7)% in the tapered shape. Linear relation was found between PCA and clinic crown length of ipsilateral central incisor in the position of CI-CI and LI-CA in square and ovoid shapes. There were significant differences in the clinic crown length of central incisors (P<0. 01), midline papilla height (PH) (P<0. 01) and gingival embrasure angle (GEA) (P<0. 01) between different shapes of central incisor of square, ovoid, and tapering. Conclusion The shapes of central incisors are closely related to the PCAP, PH and GEA, which provides reference for clinical anterior and soft tissue restoration.

[Abstract ]Objective:To estimate dental maturation norms of permanent dentition in a group of Nanjing children using Nolla's tech-nique.Methods:549 cases of panoramic radiographs (279 males and 270 females)of Nanjing children at the age range of 3 to 16 years were collected for the study.All permanent mandibular teeth on the left side (except for the third molars)were scored according to Nolla's method,and data were presented in tables and used to plot dental maturation curves for the Nanjing boys and girls.Results:Females were found to be more advanced in the degrees of dental development.Dental maturation is significantly associated with chron-ological age (for males and females:r =0.959,P <0.001 and r =0.953,P <0.001,respectively).The mathematical model of them was established,and then the fitting equation was obtained.Conclusion:The dental maturation curve and chronology of permanent dentition can provide the growth norms of permanent teeth for Nanjing children and facilitate the way for dentists to assess the growing children during diagnosis and treatment planning.

OBJECTIVEEndothelial apoptosis plays an important role in the initiation of atherosclerosis. It would be useful to clarify whether activation of non-neuronal muscarinic receptor (NNMR) could prevent endothelial apoptosis and atherosclerosis. We investigated the effects of NNMR activation on regulating rat aortic endothelial cells (RAECs) apoptosis induced by homocysteine, an independent risk factor of atherosclerosis, and further studied its molecular mechanism.

METHODSRAECs were incubated using homocysteine at the concentration of 2.7 mmol/L for 36 h. RAECs were also pre-treated with carbachol or arecoline to examine their effects. RT-PCR was used to assess changes in the gene expression related to cell apoptosis.

RESULTSIncubation of RAECs with homocysteine at the concentration of 2.7 mmol/L resulted in morphologic changes, such as cellular shrinkage, membrane blebbing, chromatin condensation and margination. These could be attenuated by pretreatment with carbachol and arecoline at the concentration of 10 micromol/L for 12 h. Homocysteine induced apoptosis in RAECs and the molecular mechanisms were associated with the regulation of fas, fas-L and caspase-8 in the death receptor pathway, bcl-2, bcl-xL and bax in the mitochondrial pathway, caspase-12 in the endoplasmic reticulum pathway and caspase-3, caspase-6 and p53 as downstream effectors. Carbachol and arecoline attenuated the effects of homocysteine on genes in the death receptor pathway, in the mitochondrial pathway and in the downstream pathway. Atropine could reverse all of the effects of arecoline.

CONCLUSIONActivation of NNMR by carbacol and arecoline inhibits homocysteine-induced endothelial cell apoptosis mainly through regulation of death receptor pathway, mitochondrial pathway and downstream effectors.

Animals ; Aorta ; cytology ; Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Arecoline ; Carbachol ; Cell Cycle ; Endoplasmic Reticulum ; metabolism ; Endothelial Cells ; cytology ; drug effects ; Homocysteine ; adverse effects ; Mitochondria ; metabolism ; Rats ; Receptors, Muscarinic ; metabolism

Objective To investigate the effect of curcumin on tumor necrosis factor-alpha (TNF-α)-in-duced expression and release of monocyte chemoattractant protein-1 (MCP-1) in rat astrocytes.Methods The primary astrocytes were prepared from the cerebral cortex of 5 neonatal Sprague-Dawley rats and cultured.The cultured cells were identified by immunofluorescence staining with glial fibrillary acid protein.The cells were then divided into 6 groups (n =15 each):control group (group C),TNF-α group (group T),TNF-α+ different concentrations of curcumin groups (Cur5,Cur10 and Cur20 groups),and TNF-α+ solvent control group (D group).TNF-α with the final concentration of 20 ng/ml was added and the cells were incubated for 2h in T group.In Cur5,Cur10 and Cur20 groups,curcumin with the final concentrations of 5,10 and 20μmol/L was added,respectively,the cells were incubated for 24h and then the culture medium was abandoned,TNF-α with the final concentration of 20 ng/ml was added and the cells were then incubated for another 2h.In group D,dimethyl sulfoxide with the final concentration of 1 μl/ml was added,the cells were then incubated for 24h,then TNF-α with the final concentration of 20 ng/ml was added and the cells were incubated for another 2h.After treatment in each group,the expression of MCP-1 was determined by immunohistochemistry and the release of MCP-1 was determined by ELISA.Results Compared with group C,the expression and release of MCP-1 was significantly increased in the other five groups (P ＜ 0.05).Compared with group T,the expression and release of MCP-1 was significantly decreased in group Cur20 (P ＜ 0.05),and no significant changes in the expression and release of MCP-1 were found in Cur5,Cur10 and D groups (P ＞ 0.05).Compared with group Cur5,the expression and release of MCP1 was significantly decreased in group Cur20 (P ＜ 0.05),and no significant change in the expression and release of MCP-1 was found in group Cur10 (P ＞ 0.05).Conclusion Curcumin can inhibit TNF-α-induced expression and release of MCP-1 in rat astrocytes and the effect is dose-related and may be one of the mechanisms of curcumin-induced reduction of neurophathic pain.

According to the TCM theory, human growth, development and aging are closely related to kidney function and the main cause of human senility is deficiency of “kidney essence”. Recent studies showed that the “kidney” in TOM include the regulatory function of the neuroendocrine and immune systems.
In this paper, the effect of Guzhen Recipe (_??__??__??_), an aging retarding prescription composed of kidney tonifying herbs such as Radix Polygoni Multiflori (_??__??_), Herba Cistanchis (_??__??_) Rubus Chiagii (_??__??__??_) etc. on the hippocampus and HPAT axis in senile rats were observed. The results showed : (1) With age the number of glucocorticoid receptors (GCR) in the hippocampus, the concentrations of NE, DA, 5-HT in the hypothalamus were markedly decreased, while the plasma corticosterone concentration in senile rats was obviously increased. (2) The thymocyte GCR nuclear transition rate was increased significantly ; (3) The contents of cytoplasmic protein, nuclear RNA and DNA, the thymus weight/body weight ratio, the serum thymic factors as well asthe rate of lymphocyte transformation, the activity of IL-2 and NK cells in senile rats were obviously decreased compared to those of young control rats. Guzhen recipe however, was capable of improving these changes in the neuroendocrine-thymic net work in senile rats to varying degrees, suggesting that the kidney with Chinese Medicine might retard the functional degeneration of thymic endocrine partly by improving the feedback regulation of the hippocampal GCR and increasing the concentrations of NE and 5-HT of hypothalamus, thus decreasing the plasma corticosterone ; Simultaneously weakening the nuclear transition rate of thymocyte GCR, which in turn improving the immune function of the senile animal.
We concluded that Guzhen Recipe may retard the functional degeneration of both the hypo-thalamuspituitary-gornadal-thymic and hypothalamus-pituitary-thyroidal-thymic axis. All the above results revealed that retarding the aging effect using TCM in volves multiple-leves, multiple-systems and links. Using holistic regulation, imbalance links in aging process are modulated and the equilibrium of Yin and Yang are maintained. This is an important characteristic of the TCM to retard aging.
This research was supported by funds from National Natural Science Foundation of China No. 38970908.

Objective To explore differences in phototest and photopatch test results, and in skin color?related parameters between healthy subjects and patients with chronic actinic dermatitis (CAD), and to examine their relationship with the melanocortin?1 receptor gene(MC1R)Arg163Gln variant. Methods Phototests were performed by using a sun simulator SUN1000, and skin color was analyzed by using Hexameter MX18 in 25 patients with CAD and 25 healthy subjects. The MC1R genotype at position?163 was determined by PCR. Photopatch tests were performed on 25 patients with CAD and 5 healthy subjects using a standard series of photoallergens(RuiMin)and an ultraviolet (UV)phototherapy equipment, SS?03A. Results Regarding phototest results, both UVA?minimal persistent pigment darkening dose(MPPD)and UVB?minimal erythema dose(MED)were significantly lower in CAD patients compared with healthy controls (both P < 0.05), with the reduction in UVB?MED being particularly notable. Sixteen patients (64%)in the CAD group had positive photopatch reactions, including 13(52%)cases of photoallergy. Skin color?related parameters were measured at four sites. Skin hemoglobin levels on the cheek, forehead, back of hands, inner upper arms were all significantly higher in CAD patients than in healthy controls(all P<0.05). However, skin melanin levels on the cheek, forehead and inner upper arms were similar between the two groups, and only those on the back of hands were significantly higher in CAD patients than in controls(P<0.01). Skin melanin and hemoglobin levels were significantly higher in exposed than in unexposed (inner upper arms) areas in CAD patients (all P < 0.05). The frequency of the CGA genotype at position?163 in the MC1R gene was similar between CAD patients and controls(P>0.05), but that of the CAA genotype differed significantly between the two groups(P<0.01). UVA?MPPD and UVB?MED were both significantly lower in CAD patients with the CAA genotype at position?163 in the MC1R gene than in those without the genotype(P=0.055, 0.325, respectively). Conclusions Skin photobiological testing plays a critical role in the diagnosis of CAD. Further studies are needed to clarify the role of the CAA genotype at position?163 in the MC1R gene in the diagnosis, prevention and treatment of CAD.

Objective The micro-vascular element plays a key role in the delivery of nutrients and the regulation of hemodynamic behavior, however, research is often hindered by ethical , economic and technological issues .Therefore, construction of a micro-vascular network in vitro will help to study the related pathological and physiological behavior in microvessels.Methods In this study, a micro-vascular element model with features of a micro-vascular network in vivo was designed based on the network structure of retinal arterioles .A micro-vascular network model in vitro, characterized by network asymmetry and the presence of both bifurcation-and side-branches , was developed by soft lithography technology . The developed microdevice allowed for the quantification of the cell -depletion layer ( CDL) thickness and hematocrit ( Ht) distribution within the microchannel networks .Results and Conclusion The study showed the potential of the developed in vitro model in revealing key hemodynamic features which have been detected for microvascular elements in vivo, including the relationships between CDL thickness , Ht and red blood cell distribution .The present study provides a new strategy and a technology for studying hemodynamics and microvascular system diseases in vitro.

Objective To investigate the role of ceramide pathway in cell proliferation and early apoptosis induction in U251 glioma cell after cannabinoid receptora agent anandmide(AEA)treatment.Methods U251 gliom cells were treated with AEA(1-10 μmol/L),Ceramide(5-20 μmol/L) and fumonisin B1 (FB1) (10 μmol/L) pretreatment.The growth inhibition rate of U251 was investigated by MTT assay.The early events of the apoptosis were measured by flow cytometry using annexin-V/propium iodide(PI) double staining method.Results Different concentrations of AEA inhibited the proliferation of human glioma U251 cells,and had synergistic effect with CM by FB1(10 μmol/L)pretreatment for 24 h.After exposure to AEA(10 μmol/L)for 24 h,U251 gliomacells could undergo the early cell apoptosis which was affected by FB1(10 μmol/L).Conclusion AEA through the CM de novo synthesis pathway,and CM concentration was lazy in collaboration,thus inhibiting human glioma U251 cell proliferation and induce early apoptosis.