Objective To estimate the influence of 1,25(OH)2D3 on the proliferative ability of and methylation levels of genomic DNA and proliferation-associated gene promoter in human HaCaT keratinocytes.Methods Some cultured HaCaT cells were treated with 1,25 (OH)2D3 of 10-6,10-7 and 10-8 mol/L for 24 hours,then,methyl thiazolyl tetrazolium (MTT) assay was carried out to evaluate the proliferative activity of cells,and a global DNA methylation quantification kit was used to determine the global DNA methylation level.Real-time PCR was conducted to quantify the mRNA expression of DNA methyl transferases (DNMTs) and methyl-DNA binding domain (MBD) proteins,and methylation-specific PCR (MS-PCR) to evaluate the methylation status of promoter region in the programmed cell death 5 (PDCD5) and tissue inhibitor of metalloproteinase 2 (TIMP2) genes,in HaCaT cells after 24-hour treatment with 1,25 (OH)2D3 of 10-6 mol/L.The HaCaT cells receiving no treatment served as the control.Results Compared with the untreated HaCaT cells,those treated with 1,25(OH)2D3 of 10-6 mol/L showed significantly down-regulated proliferative activity (0.152 ± 0.027 vs.0.290 ± 0.017,P ＜ 0.01),global DNA methylation level (0.187 ± 0.071 vs.0.316 ± 0.049,P ＜ 0.05),DNMT3a and DNMT3b mRNA expression levels (P ＜ 0.01 or 0.05),but markedly upregulated mRNA expression levels of MECP2,MBD2,PDCD5 and TIMP2 (P ＜ 0.01 or 0.05).Moreover,the DNA methylation levels within the promoter region of PDCD5 and TIMP2 genes were significantly lower in HaCaT cells treated with 1,25 (OH)2D3 of 10-6 mol/L than in the control cells (0.38 ± 0.135 vs.0.72 ± 0.121,0.46 ± 0.172 vs.0.68 ± 0.133,both P＜ 0.05).Conclusions 1,25(OH)2D3 may down-regulate the global genomic DNA methylation level of,and modulate the expression of DNA methylationmodifying genes in,HaCaT cells.Furthermore,1,25 (OH)2D3 can decrease the promoter methylation levels but induce the overexpression of PDCD5 and TIMP2 genes,and decelerate the proliferation of HaCaT cells.

Objective To investigate multi-slice spiral CT (MSCT) and MRI features of stasis cirrhosis and the diagnostic value of MSCT and MRI. Methods MSCT and MRI findings of 35 patients with stasis cirrhosis were studied. The size of liver and spleen, the diameter of hepatic vein (HV), enhancement pattern of liver parenchyma, contrast medium reflux in inferior vena cava (IVC) and (or) HV, ascites, number of varices and correlated abnormalities were reviewed. Results The volume index of liver and spleen of 35 patients was 4 434.95 cm3 and 621.92 cm3 respectively. The mean diameter of HV of 27 patients (77.1%) was 3.61 cm and HV of other 8 patients (22.9%) were too small to show. Number of patients showed waves of borderline, inhomogeneous pattern of parenchymal contrast enhancement, contrast medium reflux in IVC and (or) HV, varices and ascites was 5 (14.3%), 29 (82.9%), 20 (57.1%), 16 (45.7%), and 6 (17.1%), respectively. Correlated abnormalities included cardiac enlargement 4 cases (11.4%), pericardium thickening 11 cases (31.4%), and pericardial effusion 2 cases (5.7%). Conclusions Stasis cirrhosis mainly demonstrate liver enlargement, inhomogeneous pattern of parenchymal contrast enhancement, contrast medium reflux in IVC and (or) HV, and slight portal hypertension. MSCT and MRI play invaluable roles in diagnosis, differential diagnosis and etiological diagnosis of stasis cirrhosis.

To understand the sero-epidemiological features of rickettesiosis of humans and domestic animals in Yunnan province, blood samples from 237 adults in different geographic area, including Xundian country,Yulong country and Simao country and 81 children aged from 4 to 6 years old were collected for serological testing. In addition, 90 blood samples from dogs, goats and ox in each investigated area were also collected. Antibodies against 8 rickettsiae, including R.typhi, R.heilongjiangii or R.sibirica, Orientia tsutsugamushi Karp or Kato, Ehrlichia chaffeensis, Anaplasma phagocytosis, Bartonella henselae, Coxiella burnetii and Hainan spotted fever group of rickettsia were examined by using immunofluorescence assays(IFA).It was found that the sero-epidemiologic rates of R.typhi, B.henselae and C.burnetii (16.46%、6.33% and 9.28%) of adults were higher than those of other rickerrsiae investigated. The positive rates of IgG antibody against R.typhi for children also shared the higher rate (12.35%). Similar sero-epidemiologic features were found for domestic animals. Among the 8 rickettsia tested in this study, the positive rate of IgG antibody against R.typhi appeared to be the highest(61.48%) without significant difference among these investigated sites. From this investigation, it is evident that the rickettsial infection of farm population and domestic animals are common in Yunnan province, and the active surveillance of rickettsiosis and differential diagnosis of unknown febrile patients in clinical practice should be enforced.

Objective:To explore the effect of iron overload on the cognitive function of rats and its possible internal mechanism.Methods:Thirty 8-week-old male Sprague Dawley rats of SPF degree were randomly divided into 2 groups, iron overload group(IO group) and control group(Sham group), with 15 in each group.The rats in IO group were injected intraperitoneally iron dextran(100 mg/(kg·d)) for 28 days.The cognitive function of rats was detected by Morris water maze method. Western blot method was used to detect the expression of TfR1 and autophagy-related protein p-AMPK, LC3 and Beclin1 in the hippocampus of rats. Immunofluorescence was used to observe the expression of LC3 and Beclin1 in the hippocampus of rats. HE staining was used to observe the morphological changes of neurons in the hippocampus. Transmission electron microscopy was used to observe the number of autophagosomes and the morphology of endoplasmic reticulum in hippocampus.The software of SPSS 20.0 was used for repeated measurement ANOVA and t-test. Results:Morris water maze test showed that there were significant interaction between the group factor and training time factor of escape latency( F=3.55, P<0.01). And the simple effect analysis showed that compared with the Sham group((28.09±18.41)s, (21.42±15.53)s, (16.96±8.35)s, (10.24±3.75)s), the average escape latency of rats(2nd-5th day) in IO group((56.68±30.65)s, (58.21±36.09)s, (36.58±13.54)s, (27.29±14.30)s )were significantly longer ( t=8.57, 6.81, 9.51, 7.12, P<0.01). The platform was removed on 6th day of the space exploration experiment, compared with the Sham group ((41.89±3.89)%), the percentage of time spent in the target quadrant of IO group ((25.46±3.56)%) was significantly decreased( t=24.06, P<0.01). Western blot showed that the relative expression levels of (TfR1 (2.10±0.48), p-AMPK (0.74±0.10), LC3 (1.11±0.40), Beclin1 (1.05±0.20)) in IO group in the hippocampus of the rats were significantly higher than those of the Sham group(TfR1(0.11±0.18), p-AMPK(0.19±0.02), LC3(0.22±0.11), Beclin1(0.17±0.02))( t=1.58, 14.58, 10.06, 20.65, P<0.01)). HE staining showed that compared with the Sham group, the neuron in the hippocampus of the IO group were sparsely arranged, morphologically irregular, and the number of the neurons was significantly reduced. Transmission electron microscopy showed that compared with the Sham group, the number of autophagosomes in the hippocampus of IO group was increased. Conclusion:Iron overload may exert its neurotoxic effect by increasing the level of autophagy in the hippocampus, causing cognitive dysfunction.

Objective To analyze the possible association of psoriasis vulgaris with herpes simplex virus type 1 (HSV-1). Methods Polymerase chain reaction was used to detect HSV-1 DNA in lesional skin biopsies, periphery blood mononuclear cells (PBMCs)and throat swabs from patients with psoriasis vulgaris, and ELISA was used to detect IgM and IgG antibodies against HSV-1 in sera from these patients. Results The positive detection rates of HSV 1 DNA in lesional skin biopsies, PBMCs and throat swabs were 37.5%, 18.6%and 18.8%, respectively. Anti HSV 1 IgM and IgG antibodies were positive in 37.2%and 53.5%of serum specimens, respectively. The detection rates of HSV 1 DNA in lesional skin biopsies and PBMCs, and IgM antibody in sera were significantly higher than those in normal controls. In psoriatic patients of guttate type the positive detection rates of HSV 1 DNA and IgM antibody were significantly higher than those in the plaque type. Conclusions There is strong association of psoriasis vulgaris, especially the guttate type, with HSV 1, and there may be recent infection of HSV 1 in these patients.

Objective To fabricate an anti?tuberculosis controlled drug release coating with Ti?PDA?PEG?PLGA?INH and to investigate its surface characteristics, in vivo and in vitro drug release behavior, and tissue biocompatibility. Methods 4?arm?polyethylene glycol (PEG) was synthesized first. Then cover the surface of titanium (Ti) with a layer of poly dopamine (PDA) by Michael addition reaction. Use porous starch and 4?arm?PEG as a carrier, load with isoniazid (INH), then attach to the surface of titanium by casting or sol?gel dip coating methods, and then cover with a layer of poly lactic?co?glycolic acid (PLGA) by the same method, to fabricate the Ti?PDA?PEG?PLGA?INH composite coating finally. The functional group of 4?arm?PEG was charac?terized by proton nuclear resonance spectroscopy (HNMR). The surface characteristics of Ti?PDA?PEG?PLGA?INH were evaluated by scanning electron microscope (SEM), while drug release behaviors were detected by high performance liquid chromatography (HPLC) and the cumulative release rate was calculated, and carry out the antibacterial performance in vitro. The animal model of femoral condyle bone defect was established in 25 New Zealand white rabbits. Titanium rods covered with PDA?PEG?PLGA?INH coating were implanted into defect area. INH concentrations were detected by HPLC in venous blood, muscle and bone tissue at each time point postoperatively. Another 12 rabbits were randomly divided into experimental group and control group, the experi?mental group was implanted with titanium tablets and titanium rods coated with PDA?PEG?PLGA?INH in the paraspinous muscle and left femoral condyles respectively, while the control group was implanted with a blank sheet of titanium tablets and titanium rods in the same place. Hematoxylin and Eosin Staining were used to observe the biocompatibility of the composite system in vivo at 28 and 56 days postoperatively. Results Ti?PDA?PEG?PLGA?INH controlled drug release coating uniformly distributed on the surface of plates and rods, with translucent form and smooth surface. In vitro INH release kinetics exhibited a short?burst release during the first 8h, and the cumulative release of the INH was about 65%. On the 9th day, the cumulative release of the INH was about 90%, and then the release tended to be flat, and the drug release behavior in vitro continued more than 20d. In vivo release test showed that the concentration of INH in vein blood, muscle and bone tissue around the composite system was increased steadi?ly postoperatively. On about the 28th day, the concentration reached the max. However, the INH concentrations in muscle and bone tissue around the composite system were still higher than the minimum inhibitory concentration (MIC) on the 56th day. The antibacterial test in vitro showed that the titanium tablets coated with PDA?PEG?PLGA?INH formed obvious bacterial inhibition zones. The pathological results indicated that mild inflammatory reaction was seen in the 4th week postoperatively, and the reac?tive capsule formed with loose connective tissue. In the 8th week postoperatively, there's no obvious inflammation occurred, and the reactive capsule became more dense and thicker. Conclusion The study successfully fabricated the Ti?PDA?PEG?PLGA?INH anti?tuberculosis controlled drug release coating, with reasonable release behavior both in vivo and in vitro, effective antibac?terial effect of Mycobacterium tuberculosis in vitro and good tissue biocompatibility, which is a potentially effective drug delivery system for spinal tuberculosis.

Objective To evaluate the efficacy and safety of solifenacin in patients with overactive bladder (OAB). Methods A multicenter clinical trial was conduced. 216 patients with OAB were enrolled. All the patients received solifenacin(5 mg once daily). With 5 weeks'treatment, all the patients recorded the diary and the adverse events as well. The symptoms of urgency, frequency, nocturia, urine volume, incontinence were evaluated. The results of the efficacy and safety were analyzed by using SPSS 13. 0. Results After 5 week treatments, all the index obviously improved(P<0.05). 187 cases (86.7%)were cured and 43 cases recovered normal voiding, 29 cases improved obviously. 11cases(5.0 %)reported adverse effect as dry mouth, dry eye. Conclusion Solifenacin could be the safe and effective drug in the treatment of OAB patients.

ObjectiveTo investigate the role of hepatic stellate cell (HSC) inflammation in the pathogenesis of acute-on-chronic liver failure (ACLF). MethodsA total of 45 male Kunming mice were randomly divided into control group, model group, and N-acetylcysteine (NAC) group. The mice in the model group and the NAC group were given injection of human serum albumin to establish a model of chronic liver disease, followed by intraperitoneal injection of the endotoxins lipopolysaccharide (LPS) and D-galactosamine (D-GlaN) to induce ACLF, and those in the control group were given injection of an equal volume of normal saline; the mice in the NAC group were given NAC since 1 week before the induction of NAC. The mice in the model group and the NAC group were sacrificed at 48 hours after the injection of LPS and D-GlaN. ELISA was used to measure the serum levels of aspartate aminotransferase (AST) and alanine aminotransferase (ALT) and the levels of malondialdehyde (MDA) and superoxide dismutase (SOD) in liver tissue; HE staining was used to determine liver pathological score; ELISA was used to measure the serum levels of LPS and interleukin-1β (IL-1β). LX2 cells were stimulated by LPS and H2O2 with the presence or absence of NAC, and ELISA was used to measure the levels of IL-1β and interleukin-6 (IL-6) in medium. LX2 cells were stimulated by LPS and H2O2, and then HL7702 cells were cultured with LX2 medium; Western blot was used to measure the expression of caspase-3 and caspase-8 in HL7702 cells, and flow cytometry was used to measure the apoptosis of HL7702 cells. A one-way analysis of variance was used for comparison of continuous data between multiple groups; the least significant difference t-test was used for comparison of data with homogeneity of variance between two groups, and the Tamhane’s T2 test was used for comparison of data with heterogeneity of variance. The Kaplan-Meier survival analysis was used to evaluate survival time, and the log-rank test was used for comparison. ResultsAt 48 hours, all mice in control group survived, while 3 mice in the model group and 8 mice in the NAC group survived, suggesting that the NAC group had a better survival rate of mice than the model group (P＜0.001). Compared with the control group and the NAC group, the model group had significant increases in the serum levels of AST and ALT and the level of MDA in liver tissue, as well as a significant reduction in the level of SOD in liver tissue (all P＜0.01). The model group had a significantly higher liver pathological score than the control group and the NAC group (both P＜0.05). Both LPS and H2O2 promoted the secretion of IL-1β and IL-6 in LX2 cells, and NAC effectively inhibited the pro-inflammatory effect of H2O2 and LPS (all P＜0.05). H2O2 and LPS acted on LX2 cells and promoted the apoptosis of HL7702 cells (all P＜0.05). ConclusionLPS can promote HSC inflammation via reactive oxygen species and participates in the progression of liver failure by inducing hepatocyte apoptosis.

Objective To investigate the clinical manifestations, imaging features, treatment, and prognosis of autoimmune encephalitis （AE） with positive leucine-rich glioma-inactivated 1 （LGI1） antibody. Methods A retrospective analysis was performed for the clinical data of 11 patients with LGI1 antibody-positive AE who were admitted to Department of Neurology, The Sixth Medical Center of PLA General Hospital, from 2018 to 2022. Results Among these 11 patients, there were 10 patients with epilepsy, 8 patients with cognitive impairment, 6 patients with mental and behavioral disorders, 5 patients with sleep disorders, 1 patient with speech and language impairment, 1 patient with involuntary limb movements, 1 patient with dizziness, and 7 patients with hyponatremia. All 11 patients tested positive for LGI1 antibody, and 8 patients tested positive in serum and cerebrospinal fluid; 1 patient was also positive for contactin-associated protein-like 2 antibody, and 3 patients were positive for a single antibody in serum. Lung CT showed that 1 patient had space-occupying lesion, cranial magnetic resonance imaging showed abnormalities in 6 patients, and positron emission tomography/computed tomography showed abnormalities in 3 patients. There were 7 patients with electroencephalographic abnormalities. All 11 patients had improvements in symptoms after immunotherapy. Five patients were followed up, and 6 were lost to follow-up. Conclusion The main manifestations of LGI antibody encephalitis include seizure, faciobrachial dystonic seizures, cognitive impairment, and mental and behavioral disorders accompanied by hyponatremia. The titer of LGI1 antibody in serum is more sensitive than that in cerebrospinal fluid, and a few patients may have multiple positive autoantibodies. Immunotherapy is an effective treatment method for LGI1 antibody-positive AE.

To report a typical case of Morvan syndrome with positive anti-leucine rich glioma-inactivated 1(LGI1) and contactin-associated protein 2 (CASPR2) antibodies in serum and cerebrospinal fluid. A 39-years-old female initially presented weakness of extremeties. The main symptoms included paroxysmal limb pain, wheezing, itching, muscle twitching, epilepsy, hypomnesia, dysphoria, apathy, intractable insomnia, salivation and sweating. Tests of electrolytes found hypokalemia (2.7-3.1 mmol/L) and hyponatremia (130-136 mmol/L). Arterial blood gas analysis showed hypoxemia (oxygen saturation 50%-70%). Total thyroxine (TT4) was elevated to 207 nmol/L with positive thyroid peroxidase antibody (TPO-Ab) and thyroglobulin antibody (TG-Ab). LGI1and CASPR2 antibodies (CBA method) were positive in both serum and cerebrospinal fluid, and the remaining antibodies related to autoimmune encephalitis and paraneoplastic syndrome were negative. Head MRI was almost normal, while mild abnormalities were found in electroencephalogram. Electromyography showed slightly increased voltage of left quadriceps motor unit potential. After treated with corticosteroids, IVIG and mycophenolate mofetil, the patient completely improved. Cognitive function scores recovered from MoCA/MMSE (16/24) to MoCA/MMSE (26/29). Positivity of LGI1/CASPR2 antibodies both in serum/cerebrospinal fluid are rarely seen in patients with Morvan syndrome. Steroids and immunosuppressants are suggested for treatment as early as possible.