Objective:To investigate the expression and clinical significance of effector T cell (Teff), regulatory T cell (Treg) and their cytokines in patients with vitiligo.Methods:84 patients with vitiligo (38 cases in stable stage, 46 cases in advanced stage) who were admitted to the Dermatology Department of the Dezhou People's Hospital between June 2018 and June 2019 and 30 healthy people (control group) were enrolled in the study. The levels of Teff subsets and Treg cells in peripheral blood were detected by flow cytometry. The levels of Teff subsets and Treg cells-associated cytokines were detected by enzyme-linked immunosorbent assay (ELISA). The relationship between Teff subsets and Treg cells levels in peripheral blood, relationship between Teff subgroups, Treg cells-associated cytokines and course of vitiligo, skin lesion area were analyzed.Results:The levels of T helper cell (Th)17 and Th22 in peripheral blood of vitiligo group were significantly higher than those of control group, while levels of Treg cells were significantly lower than control group ( P<0.05). Compared with the stable vitiligo group, the levels of Th17 and Th22 in peripheral blood were higher and levels of Treg cells were lower in the advanced vitiligo group ( P<0.05). Treg cell level was negatively correlated with Th17 cell level ( r=-0.303, P=0.001), and negatively correlated with Th22 cell level ( r=-0.200, P=0.033). The levels of serum interleukin (IL)-17 and IL-22 in vitiligo group were significantly higher than those in control group, while the level of transforming growth factor-β (TGF-β) was significantly lower than that in control group ( P<0.05). Compared with the stable vitiligo group, levels of serum IL-17 and IL-22 were higher and level of TGF-β was lower in the advanced vitiligo group ( P<0.05). Correlation analysis showed that levels of IL-17 and IL-22 were positively correlated with skin lesion area ( r=0.361, 0.288, P<0.05), and level of TGF-β was negatively correlated with skin lesion area ( r=-0.312, P<0.05). Conclusions:Teff/Treg cells are unbalanced in patients with vitiligo, showing obvious shift to Teff subsets. The progression of vitiligo may be related to Teff subsets and Treg cells-associated cytokines.

ObjectiveTo investigate the effect of isoflurane preconditioning on glutamate-induced apoptosis in rat neuronal PC12 cells.MethodsThe PC12 cells were cultured for 5 d with nerve growth factor in vitro.The cells were seeded into 6-cm-diameter culture dishes (3 ml/dish) or 6-well plates (2 ml/well) with the density of 5 × 104/ml and randomly divided into 4 groups (n =18 each): normal control group (group C); glutamate group (group G) ;glutamate + isoflurane group (group GI) and glutamate + isoflurane + xestospongin C (an antngon of inositol trisphosphate receptors) group (group GIX).The neuronal PC12 cells were exposed to glutamate 500 μmol/L in groups G,GI and GIX.The neuronal PC12 cells were exposed to 1.2％ isoflurane for 2 h in groups GI and GIX.Xestospongin C was added to the culture medium immediately before isoflurane preconditioning.Glutamate was added to the culture medium at 10 min after isoflurane preconditioning in groups GI and GIX.The cells were collected from six dishes or wells in each group after being incubated with glutamate for 20 min.The apoptosis and mitochondiral membrane potential (MMP) were assessed by flow cytometry.Intracellular Ca2+ concentration ([ Ca2+ ] i)was detected by confocal fluorescence microscopy.ResultsCompared with group C,the apoptotic rate and [Ca2+ ]i were significantly increased and MMP was decreased in groups G and GIX ( P ＜ 0.01 ),but there was no significant difference in the variables mentioned above in group GI (P ＞ 0.05).Compared with group G,the apoptotic rate and [ Ca2 + ]i were significantly decreased and MMP was increased in groups GI and GIX ( P ＜ 0.05 or 0.01).Compared with group GI,the apoptotic rate and [Ca2+ ]i were significantly increased and MMP was decreased in group GIX ( P ＜ 0.01 ).ConclusionIsollurane preconditioning can inhibit apoptosis in rat neuronal PC12 cells by activating inositol trisphosphate receptors,inhibiting Ca2+ release from the endoplasmic reticulum and increasing MMP.

Objective To evaluate the role of M3 muscarinic acetylcholine receptor (M3 mAChR) in the release of glycinergic neurotransmitter by using oxotremorine-M (Oxo-M: a nonselective mAChR agonist) and 4-diphenylacetoxy-N-methylpiperidine methiodide (4-DAMP: a highly selective M3mAChR antagonist). Methods Twenty male 3-4 weeks old SD rats weighing 160-180 g after successful intrathecal catheterization were randomized into 2 groups (n = 10 each): normal saline group (group NS) and pertussis toxin (group PTX).Pertussis toxin 1.5 μg/10 μl was injected IT in group PTX, while in group NS normal saline 10 μl was injected IT instead. The animals were killed at day 7 after injection. The spinal cords were removed and sliced and placed in artificial CSF. Glycinergic spontaneous inhibitory postsynaptic currents (sIPSCs) were measured in spinal lamina Ⅱneurons using whole-cell voltage-clamp technique. Five minutes after sealing, Oxo-M (final concentration 3 μ mol/L) was added. Oxo-M was then completely washed out 3 min later and 4-DAMP (final concentration 25 nmol/L) was added after 5 min of stabilization. In the presence of 4-DAMP, Oxo-M (final concentration 3 μmol/L) was added again 3 min later. sIPSCs were recorded before addition of Oxo-M (T1), 3 min after addition of Oxo-M (T2), 3 min after addition of 4-DAMP (T3), 3 min after the second addition of Oxo-M (T4). Results Compared with the baseline value at T1 , Oxo-M significantly increased the frequency of glycinergic sIPSCs at T2without changing the amplitude at T2-4 in both groups. The frequency of sIPSCs was significantly lower at T4 than at T2 in both groups (P ＜ 0.05 or 0.01). There was no significant difference in both frequency and amplitude of glycinergic sIPSCs between the two groups. Conclusion M3 mAChR plays a predominant role in the release of glycinergic transmitter in the spinal lamina Ⅱ neurons in rats.

Objective To evaluate the role of calpain in sevoflurane anesthesia-induced apoptosis in hippocampal neurons of aged rats.Methods Fifty-four healthy female Sprague-Dawley rats,aged 18 months,weighing 450-550 g,were randomly divided into 3 groups (n=12 each) using a random number table:control group (group C),sevoflurane group (Sev group) and calpain inhibitor M DL28170 group (group M).In group C,the rats inhaled 50％ O2-50％N2 for 3 h.In Sev group,the rats inhaled 3％ sevoflurane for 3 h.In group M,MDL28170 10 mg/kg was injected via the tail vein,30 min later 3％ sevoflurane was inhaled for 3 h,and MDL28170 was simultaneously infused at 3.33 mg · kg 1 · h-1 via the tail vein.Nine rats in each group were selected,and cognitive function was assessed by using Morris water maze test at 30 min before anesthesia and 1-5 days after anesthesia.The escape latency and frequency of crossing the original platform were recorded.After the end of Morris water maze test performed at 30 min before anesthesia and 1-5 days after anesthesia,3 rats in each group were sacrificed,and hippocampal tissues were obtained for detection of cell apoptosis (by flow cytometry) and intracellular [Ca2+] i.Apoptotic rate was calculated.Results Compared with group C,the escape latency was significantly prolonged,and the frequency of crossing the original platform was decreased,and the apoptotic rate and intracellular [Ca2+]i were increased at 1 day after anesthesia in Sev and M groups.Compared with group Sev,the escape latency was significantly shortened,the frequency of crossing the original platform was increased,and the apoptotic rate was decreased at 1 day after anesthesia,and no significant change was found in intracellular [Ca2+]i in group M.Conclusion Calpain activation is involved in sevoflurane anesthesia-induced apoptosis in hippocampal neurons of aged rats.

Objective To evaluate the effect of preconditioning with nimodipine on postoperative cognitive dys function of aged rats.Methods Ninety healthy male Sprague-Dawley rats,aged 18 months,weighing 400-500 g,were randomly divided into 3 groups (n =30 each) using a random number table:nimodipine control group (group N),surgery group (group S),and nimodipine + surgery group (N+ S group).In N and N + S groups,nimodipine 1 mg/kg was intraperitoneally injected,while the equal volume of normal saline was given instead in S group.30 min later,group N inhaled pure oxygen for 2 h,and S and N + S groups inhaled 1.8 ％ isoflourane for 2 h when splenectomy was performed.Morris water maze test was performed on 1 day before operation and 1st,3rd and 7th days after operation.After the end of Morris water maze test at 1 day before operation and 1st and 7th days after operation,10 rats were sacrificed and brains were removed and hippocampi were isolated for determination of apoptosis in hippocampal neurons,intracellular [Ca2+] i in cytoplasm,and hippocampal Bcl-2 and Bax mRNA expression and for examination of ultrastructure of hippocampal neurons.Results Compared with the value before administration,the escape latency was significantly prolonged,the frequency of crossing the original platform was decreased,apoptotic rate and [Ca2+]i were increased,Bcl-2 mRNA expression was down-regulated,and Bax mRNA expression and Bax/Bcl-2 mRNA ratio were up-regulated at each time point after operation in S and N + S groups,and no significant changes were found in the parameters mentioned above in N group.Compared with group S,the escape latency was significantly shortened,the frequency of crossing the original platform was inecreased,apoptotic rate and [Ca2+]i were decreased,Bcl-2 mRNA expression was up-regulated,and Bax mRNA expression was down-regulated at each time point after operation in group N + S.Pathological changes were found in S and N + S groups and the damage was severer in S group than in N + S group.Conclusion Nimodepine preconditioning can prevent postoperative cognitive dysfunction of aged rats,and inhibition of calcium overloadinduced apoptosis in hippocampal neurons may be involved in the mechanism.

Objective To prospectively investigate the optimal setting for sentinel lymph node biopsy (SLNB) in patients with breast cancer by comparing the effects of different preparation methods and injection sites of 99Tcm-SC in sentinel lymph node (SLN) mapping and detection.Methods Two batches of 99Tcm-SC were prepared by boiling for 3 min (radiotracer 1) and 5 min (radiotracer 2),respectively.Radioactive chemical purity (RCP) and size of colloid particles were measured at 10 min,1 h,2 h and 4 h after the preparation.One hundred and forty-seven patients with breast cancer were involved and randomly divided into 3 groups.Group A consisted of 40 patients with periareolar injection of radiotracer 1,group B of 40 patients with periareolar injection of radiotracer 2,and group C of 67 patients with peritumoral injection of radiotracer 2.Lymphatic mapping was performed for all patients using SPECT/CT preoperatively and blue dye was subdermally injected over the tumor.The detection rate of the axillary and internal mammary SLN was recorded.One-way analysis of variance,independent two-sample t and x2 tests were used to analyze the data.Results There was no significant difference of RCP between the two radiotracers at 10 min,1 h,2 h and 4 h after preparation (t =-0.267,-0.794,0.826 and-0.977,all P＞0.05).Compared with radiotracer 1,the percentage of particles smaller than 100 nm in radiotracer 2 reduced significantly ((73.72±2.36) ％ vs (65.25±3.56)％,t=6.436,P＜0.05) and the mean effective particle size was significantly larger ((45.27±6.42) nm vs (75.59t7.04) nm,t=7.315,P＜0.05).In groups A,B and C,the detection rate of the internal mammary SLN was 70.0％ (28/40),47.5％ (19/40) and 17.9％ (12/67),respectively,with significant difference (x2=29.525,P＜0.05).In groups A,B and C,the detection rate of the axillary SLN was 100％ (40/40),95.0％ (38/40) and 97.0％ (65/67),respectively,without significant difference (x2 =2.686,P＞ 0.05).Conclusion For SLNB of patients with breast cancer,the axillary and internal mammary SLN could be better detected by SPECT/CT lymphatic mapping using radiotracer prepared with a shorter boiling time,via periareolar injection,and combined with subdermal injection of blue dye.

Objective To analyze the cognitive changes and influencing factors in patients with lacunar cerebral infarction after carotid artery intervention therapy. Methods Sixty lacunar cerebral infarction combined with carotid stenosis patients treated with artery intervention therapy (intervention therapy group) and 68 lacunar cerebral infarction without carotid stenosis patients treated with drug therapy (drug therapy group) were selected. The neuropsychological test was completed at entry and 1, 6, 12 months after entry, and the results were compared with 60 healthy controls (control group). The cognitive changes were observed. The neuropsychological test included mini mental state examination (MMSE), Montreal cognitive assessment scale (MoCA) and cognitive field test. Results There were statistical differences in other scores except the Stroop test C section and Wechsler adult intelligence scale (WAIS-RC) picture arrangement subtest at entry in intervention therapy group and drug therapy group compared with control group (P<0.05). There were no statistical differences in the all scores at entry between drug therapy group and intervention therapy group (P>0.05). In intervention therapy group, the MMSE scores, MoCA total score, Rey-Osterrieth complex figure test (ROCFT), auditory verb learning test (AVLT), and the WAIS-RC picture arrangement subtest, verbal fluency test, WAIS-RC digit span backwards subtest of performing function 12 months after entry were significantly better than those at entry, and there were statistical differences (P<0.05). MMSE score, MoCA total score, long-time delayed recall of ROCFT, the immediate recall, long-time delayed recall and short delayed recall of AVLT, semantic category fluency test of performing function and digit span backwards subtest of WAIS-RC 6 months after entry were significantly better than those at entry:(27.8 ± 2.2) scores vs. (26.4 ± 1.9) scores, (20.7 ± 2.3) scores vs. (19.3 ± 2.0) scores, (12.4 ± 3.2) scores vs. (10.8 ± 2.6) scores, (54.3 ± 10.6) scores vs. (49.9 ± 10.9) scores, (12.4 ± 2.0) scores vs. (11.2 ± 2.8) scores, (12.9 ± 2.0) scores vs. (10.6 ± 2.6) scores, (17.5 ± 4.0) scores vs. (15.4 ± 3.4) scores and (4.0 ± 0.9) scores vs. (3.5 ± 0.9) scores, and there were statistical differences (P<0.05). In drug therapy group, there were no statistical differences in the all scores 1 and 6 months after entry, compared with that at entry (P>0.05);the MMSE score, MoCA total score, ROCFT, the immediate recall, long-time delayed recall and short delayed recall of AVLT, WAIS-RC picture arrangement subtest, verbal fluency test, WAIS-RC digit span backwards subtest of performing function and digit span backwards subtest of WAIS-RC 12 months after entry were significantly better than those at entry, and there were statistical differences (P<0.05). There were no statistical differences in all scores 12 months after entry between intervention therapy group and drug therapy group (P>0.05). In patients intervention therapy group, Logistic regression analysis showed that the MoCA score was related with age, hypertension and low education level (P<0.01 or<0.05), but was not related with smoking, diabetes and interventional treatment (P>0.05). Conclusions Cognitive impairment in patients with lacunar cerebral infarction and carotid stenosis is severe and extensive, but most cognition disorders can improve to normal level 12 months after artery intervention therapy.

Objective To compare the anterograde amnesia produced by midazolam,propofol and dexmedetomidine when used to supplement sedation during neuraxial anesthesia.Methods Sixty patients of both sexes,aged 18-50 yr,with body mass index of 23-26 kg/m2,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,scheduled for elective operation on lower limbs with neuraxial anesthesia,were divided into 3 groups (n =20 each) using a random number table:midazolam group (group M),propofol group (group P) and dexmedetomidine group (group D).When the height of anesthesia was kept below T10,midazolam in a loading dose of O.05 mg/kg was intravenously injected in group M,propofol in a loading dose of O.4 mng/kg was intravenously injected in group P,and dexmedetomidine in a loading dose of 0.6 μg/kg was intravenously injected in group D.The infusion rate of the 3 drugs was adjusted to maintain bispectral index value at 82-86.When Observer's Assessment of Alertness/Sedation Scale scores achieved 3 or 4 after administration,anterograde amnesia was measured by postoperative recall of cards.The development of intraoperative hypotension,bradycardia and respiratory depression was recorded.Results Compared with group M,the incidence of global amnesia was significantly decreased in P and D groups (P＜0.05).There was no significant difference in the incidence of global amnesia between group P and group D (P＞ 0.05).No patients developed hypotension,bradycardia or respiratory depression in three groups.Conclusion Midazolam produces better anterograde amnesia than propofol and dexmedetomidine when used to supplement sedation during neuraxial anesthesia.

Objective To evaluate the effect of nimodipine combined with 7.5％ hypertonic saline (HS) on postoperative cognitive function in aged rats.Methods Ninety-six healthy male Wistar rats,aged 18 months,weighing 450-500 g,were assigned into 4 groups (n=24 each) using a random number table:splenectomy group (group S),nimodipine group (group N),group HS and nimodipine plus HS group (group N+HS).Nimodipine 1 mg/kg was intraperitoneally injected in group N.In group HS,7.5％ HS 4 ml/kg was injected via the caudal vein.The equal volume of normal saline was injected intraperitoneally or via the caudal vein in group S.Splenectomy was performed under sevoflurane anesthesia at 30 min after the end of administration.On 1 day before operation and 3 and 7 days after operation,Morris water maze test was performed,and blood sainples from the caudal vein were simultaneously collected for determination of the concentrations of serum S100β protein and neuron-specific enolase (NSE) by enzyme-linked immunosorbent assay.Results Compared with group S,the frequency of crossing the original platform was significantly increased,the escape latency was shortened,and the concentrations of serum S100β protein and NSE were decreased at each time point after operation in N,HS and N+HS groups (P＜0.05).Compared with group N or group HS,the frequency of crossing the original platform was significantly increased,the escape latency was shortened,and the concentrations of serum S100β protein and NSE were decreased at each time point after operation in group N+HS (P＜0.05).Conclusion Nimodipine combined with 7.5％ HS exerts better efficacy than either alone in improving postoperative cognitive function in aged rats.

Objective:To study whether the "combination of medical care and pension" service model can effectively control the development of chronic disease in the elderly and reduce the direct economic burden caused by the disease.Methods:A total of 180 elderly participants who received the "combination of medical care and pension" service model for chronic disease management in Chongqing, China were investigated and analyzed statistically. Epidata 2.0 was used for data entry, SPSS 20.0 was used for data analysis, and paired sample t test was used for comparison between groups. Results:After 12 months of chronic disease management, hospitalization events and expenses of the elderly were reduced, among which the number of hospitalization was reduced by 0.24 timed on average, the length of hospitalization was reduced by 10.41 days on average, and the hospitalization expenses were reduced by 11 144 yuan on average. The direct economic burden due to chronic diseases was reduced by approximately 8 844.5 yuan, accounting for 33.8% of the average cost of hospitalization for the elderly without the application of the model.Conclusion:The chronic disease of the elderly is well controlled by chronic disease management through the "combination of medical care and pension" service model.