Objective • To investigate whether methylene blue-mediated photodynamic therapy (MB-PDT) can induce the apoptosis of macrophages in periodontitis, and simultaneously reduce the bone absorption. Methods • For in vitro treatments, cells were divided into three experimental groups: control group, no treatment; MB group, methylene blue treatment; MB-PDT group, MB and laser irradiation treatment. Then apoptosis and apoptosis related genes were detected in each group. For in vivo treatments, periodontal disease in SD rats was orthodontic ligature and periodontal pathogen induced at the first maxillary molar. After 6 weeks, the ligature was removed and all animals received scaling and root planning(SRP) and were divided according to the following treatments: SRP group, saline solution; MB group, phenothiazinium dye; and MB-PDT group, MB and laser irradiation, once a week. All animals in each treatment were killed after 3 weeks. Immunofluorescence and Micro CT analyses were used to detect the apoptotic macrophages and alveolar bone resorption in periodontal tissues of rats in each group. Results • In vitro experiments showed that the combination of 10 μmol/L MB and 40 J/cm2 light dose could kill more than 50% macrophages, and the apoptosis of macrophages induced by MB-PDT in this mode was most obvious. Meanwhile, MB-PDT increased the expression ratio of proapoptotic gene Bax /antiapoptotic gene Bcl-2 in macrophages. In vivo experiments showed that the macrophages infiltrated in tissues with periodontitis were apoptotic after MB-PDT treatment, while no obvious apoptosis of macrophages infiltrated in the periodontal tissues of SRP and MB rats was found. Micro CT analysis showed that the alveolar bone resorption in MB-PDT rats was less than that in SRP and MB rats (P<0.05), and there was no significant difference between SRP rats and MB rats. Conclusion • MB-PDT can induce the apoptosis of hyperproliferating macrophages in periodontitis and reduce the bone absorption. Compared with SRP, MB-PDT is an effective adjunctive treatment of periodontitis.

Adult ; Angiomyolipoma ; metabolism ; pathology ; surgery ; Carcinoma, Renal Cell ; metabolism ; pathology ; Diagnosis, Differential ; Humans ; Kidney Neoplasms ; metabolism ; pathology ; surgery ; MART-1 Antigen ; metabolism ; Male ; Melanoma-Specific Antigens ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

Objective:To compare and explore the effects of needling acupoints at different nerve segmentson the oxytocin (OT) neurons in the paraventricular nucleus of hypothalamus (PVN) and the intragastric pressure, and discuss the possible mechanisms. Methods: Thirty-two healthy adult Sprague-Dawley (SD) rats were numbered and divided into 4 groups according to the random number table, a Zusanli (ST 36) group, a Neiguan (PC 6) group, a Weishu (BL 21) group and a control group, with 8 rats in each group. Except the control group, rats in the other three groups received acupuncture at the corresponding acupoints. To observe the differences in double-labeled OT neurons and c-fos neurons of the hypothalamic PVN and the intragastric pressure after acupuncture among the three groups of needling acupoints at different nerve segments. Results:Compared with the control group, the numbers of double-labeled cells in the PVN of the Zusanli (ST 36) group and the Neiguan (PC 6) group decreased significantly, while the intragastric pressure increased significantly (allP<0.05), and the inter-group differences were statistically significant (P<0.05). The intragastric pressure in the Weishu (BL 21) group decreased significantly, and the inter-group difference was statistically significant (P<0.05). Compared with the Weishu (BL 21) group, the numbers of OT/c-fos double-labeled cells in PVN of the Zusanli (ST 36) group and the Neiguan (PC 6) group decreased significantly, and the intragastric pressure increased significantly, the inter-group differences were statistically significant (allP<0.01). Conclusion:Acupoints at different nerve segments have different regulation effects on intragastric pressure. The difference may be related to the different nerve conduction pathways by acupoints at different nerve segments in regulating the intragastric pressure. The PVN may be one common integration center for the regulation of gastric function in the three acupoints [Zusanli (ST 36), Neiguan (PC 6) and Weishu (BL 21)] at different nerve segments.

Adult ; Female ; Humans ; Ovarian Diseases ; diagnosis ; Ovarian Neoplasms ; diagnosis ; Pregnancy

The present study was undertaken to investigate the effects of ceramide on progesterone production and apoptosis in rat luteal cells in vitro. Luteal cells were prepared from the ovaries of pregnant mare serum gonadotropin (PMSG) and human chorionic gonadotropin (hCG) primed female Wistar rats and incubated with cell permeable C2-ceramide. The effects of ceramide on progesterone production and apoptosis in luteal cells were assessed by radio immunoassay (RIA) and flow cytometry analysis. In addition, changes in nitric oxide synthase (NOS) activity and nitric oxide (NO) production by luteal cells treated with C2-ceramide were also evaluated. Ceramide was found to reduce hCG-stimulated progesterone production in a dose-dependent manner, whereas it had little effect on basal progesterone content. Spontaneous apoptosis in luteal cells was observed after a 12-hour incubation in vitro and 5 μmol/L of ceramide significantly increased the apoptotic rate (P<0.05). Enhanced apoptotic peak was seen in the histogram by flow cytometry. Moreover, 50 μmol/L of C2-ceramide significantly increased NOS activity (P<0.01) and NO production (P<0.001). It is suggested that ceramide may serve as an important signaling molecule mediating certain ovarian processes, such as luteal regression.

Objective To observe the advantages and disadvantages the combination of traditional Chinese medicine and western medicine in community health service centers through cost-effectiveness and cost-utility analysis.Methods Take community common diseases as an example, Knee Osteoarthritis (KOA), Cervical Spondylosis (CS) and Lumbar Disc Herniation (LDH) were used. A total of 200 cases were selected for each disease, and 100 cases in each one group.In the treatment group, the rehabilitation mode was used.In the control group, comprehensive treatment of traditional Chinese medicine was adopted. We analyzed the cost-effectiveness and cost-utility, with VAS as the effect indicator and QALYs as the utility indicator. Results The VAS values of the knee osteoarthritis, cervical spondylosis and lumbar disc herniation in the treatment group were lower than those in the control group after 14 days, 30 days and 6 months (P<0.05) . The QALYs values of them in the treatment group were lower than those in the control group after 14 days, 30 days and 6 months (P<0.05) . The treatment group has a higher increase cost-effectiveness ratio (ICER) . Each get a unit effect can pay 222.56, 297.13 and 1178.59 Yuan less . The treatment group has a higher increase cost-utility ratio (ICUR) . Each get a unit effect can pay 741.86, 3178.60 and 2862.29 Yuan less . Conclusion Community application in rehabilitation mode of integrated traditional Chinese and western medicine can improve the treatment effect, and can reduce the cost of treatment and the loss of QALYs.

BACKGROUNDRetroperitoneal fibrosis (RPF) is an uncommon disease that is characterized by development of fibrosclerotic tissues involving retroperitoneal structures. This study aimed to investigate the clinical features of 30 patients with RPF in a single center in Beijing in a 10-year period.

METHODSWe retrospectively analyzed clinical data on demographic characteristics, clinical manifestations, laboratory findings, radiological findings, modalities of treatments, outcomes and prognosis of 30 patients with RPF. Patients were treated in Beijing Chao-Yang Hospital between January 2003 and December 2013.

RESULTSThe mean age of patients with RPF was 56.7 ± 14.4 years. Twenty-three patients were men and seven patients were women. Acute phase reactants were elevated in most patients. Rheumatic factor was positive in 4/25 (16.0%) patients, and antinuclear antibody was positive in 6/22 (27.3%) patients. Elevation of IgG4 was observed in 9/22 (40.9%) patients. The most common type was I + III (n = 13), followed by I + II + III (n = 12). Five patients undertook an 18 F-fluoro-deoxy-D-glucose positron emission tomography examination and increased uptake was detected in four patients. Eight patients received combination therapy with glucocorticoids and tamoxifen. Surgical intervention treatments included intraureteral double-J stent implantation (n = 26), percutaneous nephrostomy (n = 2), open ureterolysis and intraperitonealization of the ureters (n = 5) and laparoscopic ureterolysis and intraperitonealization of the ureters (n = 5). Three patients underwent hemodialysis because of renal failure.

CONCLUSIONSClinical characteristics of RPF patients in our study are similar to those previously reported. Steroids and immunosuppressive therapy combined with ureterolysis could be a viable choice of treatment for RPF. More prospective, multi-center studies with a longer follow-up are warranted.

Adult ; Aged ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Immunoglobulin G ; blood ; Male ; Middle Aged ; Retroperitoneal Fibrosis ; blood ; diagnosis ; drug therapy ; surgery ; Retrospective Studies ; Tamoxifen ; therapeutic use ; Treatment Outcome

OBJECTIVETo investigate mutations of anaplastic lymphoma kinase (ALK) in Chinese children with neuroblastoma (NB).

METHODSGenomic DNA was extracted from 22 cases of paraffin-embedding NB tumor tissues. Gene mutations in the exons 20-26 which were mutational hotspots of ALK were analyzed by PCR-DNA direct sequencing.

RESULTSA novel synonymous mutation C3586T (Leu1196Leu) and a known synonymous mutation C3375A (Gly1125Gly) were found and located at exon 23 and exon 21 of ALK respectively. There were 10 cases (46%) of known synonymous mutation C3375A in 22 cases of NB. The C3375A allelic frequency was 27%. No statistically significant correlation was found between mutation C3375A and clinical parameters of NB such as age, sex, metastasis and tumor differentiation. Mutation was not found in the other 5 exons.

CONCLUSIONSA novel ALK gene synonymous mutation C3586T was identified using PCR-DNA sequencing. A known mutation C3375A in ALK was successfully identified in children, and its incidence is not influenced by the clinical features of childhood NB.

Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Mutation ; Neuroblastoma ; genetics ; Polymerase Chain Reaction ; Receptor Protein-Tyrosine Kinases ; genetics

OBJECTIVETo look for the evidences of motilin receptor expression on interstitial cells of Cajal (ICC) of the rabbit.

METHODSmooth muscle segments with ICC were isolated from the small intestine of 10-day old rabbits. The tissue segments equilibrated in Ca(2+)-free Hanks' solution were dispersed with an enzyme solution containing collagenase type II and then Ficoll density centrifugation was used to dissociate ICC. The cells were suspended and cultured in the M199 medium. The c-kit antibody was applied to distinguish the cultured ICC. The motilin receptor was identified by immunocytochemical assay with GPR38 antibody, c-kit antibody and hoechst 33342 combined to label ICC. Cells cultured for a few days were sorted for ICC with c-kit stained green fluorescent through flow cytometry. The total RNA and proteins extracted from the sorted ICC were respectively used to verify motilin receptor on the ICC by reverse-transcriptase polymerase chain reaction (RT-PCR) and Western blotting.

RESULTWe had successfully dissociated and cultured ICC of rabbit small intestine in vitro. Fluorescent staining with c-kit antibody confirmed that the culture ICC was successful. Triple-labeled immunofluorescent staining had detected the motilin receptor on membrane of ICC. Flow cytometry analysis showed that the ratio of c-kit positive cell in the cultured cells was 64.3%. The number of sorted ICC was 6.7 × 10(5) and 5.6 × 10(6). The results of RT-PCR and Western blot confirmed that the ICC had motilin receptor expression.

CONCLUSIONOur study demonstrated presence of motilin receptor on ICC of the rabbit. The present results may suggest that ICC play an important role in gastrointestinal movement induced by motilin.

Animals ; Cells, Cultured ; Interstitial Cells of Cajal ; metabolism ; Intestine, Small ; cytology ; Rabbits ; Receptors, Gastrointestinal Hormone ; metabolism ; Receptors, Neuropeptide ; metabolism

AIMTo study the metabolite of stilbene glucoside in the Chinese traditional medicine Polygonum multiflornm in mice and elucidate its chemical structure by liquid chromatography tandem-mass spectrometry.

METHODSThe stilbene glucoside was injected into the tail vein of mice. Blood samples were collected from artery in the eyepit. The methanol-protein-precipitated plasma sample was introduced into the liquid chromatography-tandem mass spectrometer directly. The analytical column was C18 column (250 mm x 4.6 mm ID, 5 microns). The mobile phase consisted of acetonitrile-methanol-water-formic acid (15:18:66:1) for ES+, acetonitrile-methanol-water (15:18:67) for ES-. The UV detection wavelength was set at 320 nm. The mass ion source type was ESI. HV capillary was 3 kV. The dry gas was nitrogen gas and the flow rate was set at 318 L.h-1. The ion source temperature was 150 degrees C.

RESULTSThe stilbene glucoside and its metabolite were separated completely under the chromatography condition. The ions at m/z 600 and m/z 605 were detected under positive ion polarity while the ions at m/z 581 and m/z 402 were detected under negative ion polarity.

CONCLUSIONIt was proposed that the metabolite of stilbene glucoside injected in vein was its glucuronide conjugate.

Animals ; Chromatography, Liquid ; methods ; Female ; Glucosides ; blood ; isolation & purification ; metabolism ; Male ; Mice ; Plants, Medicinal ; chemistry ; Polygonum ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Stilbenes ; blood ; chemistry ; isolation & purification ; metabolism