Background Retinal microvascular pericytes (RMPs) have been played increasing attention as an emerging key in pathogenesis of various retinal angiogenic diseases including diabetic retinopathy,and RMPs are thought to be a potential target for treatment.Yet the study has been hindered by the difficulty of obtaining source of tissue and isolating pure population.Objective This study was to establish a simple method of isolation,purification and cultivation of primary RMPs for rat.Methods Eyeballs were extracted from clean male Sprague Dawley rats and immersed by 75％ alcohol for 1 minute.The retinas were isolated and mechanical morcel.Trypsin (2.5 g/L) was firstly used and followed by type Ⅰ collagenase (2 g/L) for the digestion of the retina for 15 minutes,respectively.Retinal microvascular fragments were screened by 100 μm and 55 μm filter screen.DMEM containing 20％ fetal bovine serum was added for the cultivation and passaged of the cells.The cells were purified by exchanging medium and partial enzymatic digestion.The morphology and growth status were monitored under the phase contrast microscope,and α-smooth muscle actin (α-SMA),platelet-derived growth factor receptor-β (PDGFR-β),yon Willebrand factor (vWF),glial fibrillary acidic protein (GFAP) antibodies were used for the identification of RMPs.Results RMPs migrated out of fragments after 24-48 hours of plating.On day 7,RMPs appeared in primary cultures as loose colonies.The cells reached confluence to about 80％-90％ on day 14-16.The subcultures grew faster than the primary and reached confluence on day 12-14.The culture showed typical morphology of pericyte with large irregular triangular cell body and multiple long processes,and they could be repeatedly passaged 9 times without obvious loss of characteristic phenotype.Fluorescence assay exhibited that 96％ of the cells showed positive immunofluorescence for α-SMA and PDGFR-β,confirming the purity of RMPs in culture.However,only a few of them were positive for GFAP and the cells response for vWF was absent.Conclusions High purity of rat RMPs can be obtained easily by our method without high cost-consuming.Hcrc wc cstablished a simple mcthod for the primary culture of rat RMPs.

Objective To analyze the clinical factors predicting long-term survival after curative resection of pancreatic head carcinoma. Methods The clinical data of 58 patients with ductal adenocarcinoma of pancreatic head who underwent curative resection of carcinoma of pancreatic head from 1996 to 2004 were collected and were analyzed by SPSS 10.0 with Cox Proportional Hazards Model. Results 58 patients, including 30 male and 28 female patients, were involved in this study. Pancreaticoduodenectomy were performed in 14 cases and extended resections were performed in 44 cases. The overall 1, 3, 5 year survival rates was 46.6%, 29.3% and 8.6%, respeclively. The 1,3, 5 year survival rates of pancreatoduodenectomy with extended regional lymphadenectomy was 43.1%, 22.7% and 6.8%, respectively. UICC staging, peri-pancreatic nerve invasion and blood infusion had significant effects on the prognosis after curative resection. Conclusions The long-term prognosis after curative resection of pancreatoduodenectomy was still dismal. Much importance should be paid to early diagnosis and comprehensive management for pancreatic head cancer.

OBJECTIVETo evaluate the optimal insertion position of the Coflex lumbar interspinous dynamic stabilization device.

METHODSSix fresh adult human cadaveric lumbar spine specimens (L1-L5) were mounted in a materials testing machine by embedding to clamps with L1 and L5 vertebrae. L3-4 motion segment of each specimen was operated by selective decompression and Coflex interspinous device insertion. The L3 and L4 vertebrae was inserted one needle attached with four marker points respectively, which were used to record the range of motion (ROM). Each lumbar spine specimen was tested according to the loading sequence at 5 groups: intact (keeping lumbar ligamenta and facet joints intact) group, partial destabilized (resection of L3-4 interspinous ligamenta, ligamentum flavum, facet capsule, and bilateral resection 50% of L3 inferior facets) group, 10 mm insertion (distance between apex of U-shaped Coflex and dural sac was 10 mm)group, 5 mm insertion (distance was 5 mm)group, and 0 mm insertion (distance was 0 mm)group. Each lumbar spine specimen was tested repeatedly 3 times according to a loading sequence consisting of flexion, extension, left/right lateral bending, left/right axial rotation, loaded with pure moments of 8 N·m, and was recorded the ROM of operative segment at the third time. ROM of 5 groups in 6 directions respectively were analyzed with one-way ANOVA test and multiple comparisons were based on LSD method.

RESULTSThe means ROM of 5 groups were not all equal in flexion, extension, left/right lateral bending, left/right axial rotation (F = 8.472, 18.301, 7.700, 12.473, 16.809, 6.624; all P < 0.01). The 10 mm insertion group had significant high ROM in 6 directions than the intact group (t = 3.80, 3.82, 4.49, 5.60, 4.96, 2.98, all P < 0.01), but it was no difference comparing with the partial destabilized group (P > 0.05). The ROM of the 5 mm and 0 mm insertion group were no significant differences comparing with the intact group in flexion, extension, left/right axial rotation (P > 0.05), but it were significant differences comparing with the partial destabilized group in the same directions (5 mm insertion group: t = 3.19, 6.34, 5.26, 3.43, all P < 0.01; 0 mm insertion group: t = 4.21, 6.68, 5.81, 3.72, all P < 0.01). There were significant differences in the ROM of left/right lateral bending between the 5mm/0mm insertion groups and the intact group (5 mm insertion group: t = 3.71 and 5.22, all P < 0.01; 0 mm insertion group: t = 3.44 and 4.95, all P < 0.01), but there were no differences comparing with the partial destabilized group in the same directions (P > 0.05).

CONCLUSIONSThe insertion of Coflex interspinous dynamic stabilization device can maintain the stability of a partially destabilized specimen back to an intact one in flexion, extension and axial rotation when distance between apex of U-shaped Coflex and dural sac was ≤ 5 mm, but can't return the stability in lateral bending. The Coflex can't return the stability of a partially destabilized specimen back to an intact one in 6 directions when distance between apex of U-shaped Coflex and dural sac was ≥ 10 mm.

Adult ; Biomechanical Phenomena ; Bone Screws ; Decompression, Surgical ; Humans ; Internal Fixators ; Ligamentum Flavum ; surgery ; Lumbar Vertebrae ; surgery

Objective: To investigate the clinicopathological characteristics of reactive epithelial hyperplasia and dysplasia in the stomach, as well as the clinical value of mucin special staining and proliferating cell nuclear antigen (Ki-67) in distinguishing the two gastric lesions. Methods: The clinical pathological data of 63 patients with gastric reactive epithelial hyperplasia, 54 patients with low-grade dysplasia, and 63 patients with high-grade dysplasia diagnosed from May 2018 to May 2021 in Beijing Chaoyang Hospital, Capital Medical University, Beijing, China were analyzed. Alcian blue periodic acid Schiff (AB-PAS) and Ki-67 staining were performed to examine the mucin staining pattern, number of Ki-67 positive cells, Ki-67 staining patterns in the three groups of lesions, and histopathologic characteristics. Results: The positive rates of AB-PAS in the reactive epithelial hyperplasia and gastric dysplasia groups were 87.3%(55/63) and 10.3%(12/117), respectively. The expression of AB-PAS in the reactive epithelial hyperplasia was gradually increased from the base to the surface of the epithelium. In low-grade dysplasia and high-grade dysplasia, there was no mucin present in the dysplasia epithelium. The difference between the two groups was statistically significant (P<0.01). The positive rate of Ki-67 in the epithelial reactive hyperplasia (>10%) was 81.0% (51/63), and the positive cells were mainly located in the neck and middle parts of the mucosal glands (58/63, 92.1%). In the low-grade dysplasia group, the positive rate of Ki-67 (>10%) was 90.7%(49/54); the positive cells were mainly located in the upper mucosa (33/54, 61.1%), showing a banded distribution pattern; in the high-grade dysplasia group, the positive rate (>10%) was 95.2%(60/63), and the positive cells were mainly located in the whole mucosa (49/63, 77.8%), showing a diffuse/diffuse scattered distribution pattern. The three groups had statistically different rates and distribution patterns of Ki-67 expression (P<0.01). Conclusion: The gastric epithelial reactive hyperplasia and dysplasia can be differentiated using clinicopathological features, AB-PAS staining and Ki-67 expression pattern.
Alcian Blue ; Humans ; Hyperplasia ; Ki-67 Antigen/metabolism* ; Periodic Acid ; Staining and Labeling ; Stomach Neoplasms/diagnosis*

OBJECTIVEThis study aims to investigate relationship between daily concentration of PM(10), SO(2), NO(2) and daily mortality due to circulatory system diseases in Chaoyang district, Beijing.

METHODSThe time-series data of daily mortality from circulatory system diseases of registered residents in Chaoyang were obtained from Chaoyang District Center for Disease Control and Prevention. The daily concentration of sulfur dioxide (SO(2)), nitrogen dioxide (NO(2)), and particulate matter (PM(10)) were collected from Beijing Municipal Environmental Monitoring Center. And the routine monitoring meteorological data were collected from Beijing Meteorological Bureau, including daily mean temperature and daily mean relative humidity. The time-series analysis was then conducted to determine the relationship of mortality from circulatory system diseases with daily concentrations of SO(2), NO(2) and PM(10) by using Poisson regression with generalized additive model (GAM).

RESULTSDuring January 2004 to September 2008, the cumulative death number from circulatory system diseases of registered residents in Chaoyang district of Beijing was 19 241, the daily average concentration of SO(2), NO(2), PM(10) was 48.7, 63.9, 146.1 µg/m(3), respectively. The single pollutant model showed an increase of daily concentration of PM(10), SO(2) and NO(2) by 10 µg/m(3) will augment the mortality from circulatory system diseases by 0.20% (95%CI: 0.01% - 0.39%), 0.36% (95%CI: -0.13% - 0.85%) and 0.30% (95%CI: -0.34% - 0.94%), respectively. In the multiple air pollutants models, combinatorial effects of PM(10) and SO(2) still positively correlated with increased mortality from circulatory system diseases (P < 0.05), whereas changes of the concentration of NO(2) had no significant effect on mortality from circulatory system diseases (P > 0.05).

CONCLUSIONOur findings in this study elucidated that changes of the concentration of PM(10) and SO(2) had a positive correlation with daily mortality from circulatory system diseases among the local residents in Chaoyang District, whereas the daily concentration of NO(2) was irrelevant with that.

Aged ; Air Pollutants ; analysis ; Air Pollution ; analysis ; Cardiovascular Diseases ; epidemiology ; mortality ; China ; epidemiology ; Environmental Monitoring ; Female ; Humans ; Male ; Models, Theoretical ; Nitrogen Dioxide ; analysis ; Particulate Matter ; analysis ; Sulfur Dioxide ; analysis ; Survival Rate ; Time Factors

BACKGROUNDSeveral candidate genes of ossification of the posterior longitudinal ligament (OPLL) susceptibility have been identified, but their polymorphisms account for only a small percent of the total variance. Bone morphogenetic protein-4 (BMP4) is a potent ectopic ossification inducing factor. BMP4 protein and mRNA are present in cells from OPLL patients, but not non-OPLL controls. A single nucleotide polymorphism of 6007C>T(rs17563) of BMP4 has been reported to affect bone density in postmenopausal women. Thus, BMP4 may function in OPLL development. Appropriately, the relationship between BMP4 polymorphisms and OPLL was investigated.

METHODSA case-control association study investigated the genetic etiology in 179 OPLL patients and 298 non-OPLL controls. Extent of OPLL was analyzed by radiologic examinations. Whether single nucleotide polymorphism (SNP) of -5826G>A(rs1957860) 5' of the transcription start site and 6007C>T(rs17563) in exon 4 of the BMP4 gene were statistically associated with genetic susceptibility to OPLL in Chinese Han subjects was assessed.

RESULTSA significant statistical difference in genotype of 6007C>T polymorphism between male OPLL patients and male controls was evident, and the frequency of "TT" genotype in male OPLL patients was significantly higher than in male controls (P = 0.039). The frequency of the "T" allele was also significantly higher in male OPLL subjects than in male controls (P = 0.014, OR = 1.57). A significant difference was also observed between the 6007C>T polymorphism and the number of ossified cervical vertebrae in OPLL patients, while no statistical difference was apparent between the -5826G>A polymorphism and OPLL occurrence.

CONCLUSIONSThe T allele in the 6007C>T polymorphism may be a risk factor for male Han Chinese with ossification of the posterior longitudinal ligament in the cervical spine. Chinese Han male patients with CT and TT 6007C>T genotypes have a genetic susceptibility to OPLL and more extensive OPLL in the cervical spine.

Alleles ; Asian Continental Ancestry Group ; genetics ; Bone Morphogenetic Protein 4 ; genetics ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Male ; Middle Aged ; Ossification of Posterior Longitudinal Ligament ; genetics ; Polymorphism, Single Nucleotide ; genetics

Objective: To study the clinical results of Coflex and lumbar posterior decompression and fusion in the treatment of lumbar degenerative spondylolisthesis at L_4-5. Methods: Thirty-eight patients with Grade Ⅰ degenerative spondylolisthesis, from January 2008 to December 2011 in Beijing Chaoyang Hospital, Capital Medical University were reviewed, and patients were divided into two groups by randomness. Group A was treated with Coflex and group B with pedicle instrumentation and interbody fusion. Fifteen patients were included in group A, and 23 patients were included in group B. In group A, the average age was (56.3±9.1) years. In group B, the average age was (58.2±11.2) years. The clinical results were evaluated by visual analogue scale (VAS) and Oswestry disability index (ODI). Slip distance (SD) was measured before and after surgery, and the changes of intervertebral angle at index level and adjacent level were also recorded. Results: The follow-up period was 36 to 68 months, with the average of (39±14) months in the both groups. The operation time and bleeding volume of patients in group A were significantly less than that of group B (P<0.05). In both groups, the difference of ODI and VAS before operation and postoperative follow-up were statistically significant (P<0.05). There was no significant difference between lumbar intervertebral angle and the sliding distance in group A at all time points. In the group B, there was a significant increase in the intervertebral angle and the sliding distance at L_3-4 and L₅-S₁ level after surgery, the difference at upper and below adjacent segment before and after surgery were statistically significant. Conclusions Coflex interspinous dynamic stabilization system has same excellent clinical results as pedicle screw instrumentation and fusion surgery for the treatment of L_4-5 degenerative spondylolisthesis; no significant progression of spondylolisthesis been observed during more than 3 years follow-up, and no obvious adjacent segment degeneration has been found.

Objective: To analyze the risk factors of implant-related complications in adult patients with adult degenerative scoliosis (ADS) who underwent long-level internal fixation. Methods: This was a retrospective study that analyzed 99 cases of adult degeneration scoliosis patients who underwent long level posterior pedicle screw fixation in our hospital between June 2013 and January 2016. The internal fixation and related complications were evaluated by measuring and analyzing the radiographic data of the postoperative X-ray films. The timepoints of measurement were pre-operation, before discharge, half a year, 1 year, when complications occur and the final follow-up after operation. Implant-related complications included proximal junctional kyphosis (PJK), proximal junctional failure (PJF), distal junctional kyphosis/failure (DJK/DJF), rod breakage in addition to other radiographic implant-related complications (that were not related to PJF) such as screw loosening, breakage or pullout, or interbody graft and hook or set screw dislodgements seen on follow-up radiographs. The incidence of postoperative implant-related complications was counted. All the cases included in the study were divided into the complication group and the non-complication group. Statistical difference between groups at different follow-up time point was analyzed. Potential risk factors were identified using univariate testing. Multivariate Logistics regression was used to analyze the independent risk factors for implant-related complications. The postoperative functional scores were assessed using the Oswestry Disability Index (ODI), Japanese Orthopaedic Association Scores (JOA), Visual Analogue Scale (VAS), and Lumbar Stiffness Disability Index (LSDI). Functional scores were tested using group t tests. Patients were divided into groups according to PI-LL <10 °, 10°-20 ° and > 20 °. The preoperative and postoperative radiographical parameters and clinical function score among each groupwere compared.The best PI-LL matching value was verified by analyzing the effect of long-segment fusion orthopedics on ADS. Results: Ninety-nine ADS patients who underwent long level posterior fixation were included. The incidence of patients with mechanical complications was 30.3%. Univariate analysis showed that chronic risk factors of postoperative implant-related complications after surgery of ASD included diabetes (OR=3.52, P=0.001) and blood transfusion (OR=2.61, P=0.030); surgical risk factor isosteotomy (OR=4.33, P=0.000); preoperative imaging risk factor was preoperative SVA (OR=1.03, P=0.000); the risk factors for increased risk of the implant-related complications included anemia (OR=1.17, P=0.810), cardiac complications (OR=1.80, P=0.290) and hospital stay (OR=1.11, P=0.110). Independent predictors identified on multivariate Logistics regression modeling included osteotomy (OR=3.05, P=0.032), and preoperative SVA (OR=1.03, P=0.007). The radiographical parameters and clinical function scores of the PI-LL 10°-20° group were better than or partially superior to those of the PI-LL<10° group and the PI-LL>20° group. The postoperative SVA of the PI-LL 10°-20° group was significantly lower than that of the PI-LL<10° group (t=2.399, P=0.020) and the PI-LL>20° group (t=-3.074, P=0.005). The incidence of implant-related complications in the PI-LL 10°-20° group was significantly lower than that in the PI-LL<10° group (t=1.584, P=0.003). Survival analysis showed that the PI-LL 10°-20° group was significantly better than the PI-LL<10° group (χ²=7.782, P=0.005), while the PI-LL 10°-20° group had better survival than PI-LL>20° group, althoughthatwas not statistically significant (χ²=2.542, P=0.111). Conclusion Risk factors of postoperative implant-related complications after surgery of ASD included osteotomy and preoperative SVA. Patients with one or more of these risk factors should be informed of the risk increase with informed consent. Patients with PI-LL between 10 ° and 20 ° had better postoperative radiographical parameters and clinical functional scores. They should be optimized preoperatively and followed up closely during the postoperative period.

BACKGROUND: Spinal cord injury (SCI) is a worldwide medical concern. This study aimed to elucidate the mechanism underlying the protective effect of hyperbaric oxygen (HBO) against SCI-induced neurologic defects in rats via exploring the stromal cell-derived factor-1 (SDF-1)/CXC chemokine receptor 4 (CXCR4) axis and expression of brain-derived neurotrophic factor (BDNF). METHODS: An acute SCI rat model was established in Sprague-Dawley rats using the Allen method. Sixty rats were divided into four groups (n = 15 in each group): sham-operated, SCI, SCI treated with HBO (SCI + HBO), and SCI treated with both HBO and AMD3100 (an antagonist of CXCR4; SCI + HBO + AMD) groups. The rats were treated with HBO twice a day for 3 days and thereafter once a day after the surgery for up to 28 days. Following the surgery, neurologic assessments were performed with the Basso-Bettie-Bresnahan (BBB) scoring system on postoperative day (POD) 7, 14, 21, and 28. Spinal cord tissues were harvested to assess the expression of SDF-1, CXCR4, and BDNF at mRNA and protein levels, using quantitative real-time polymerase chain reaction, Western blot analysis, and histopathologic analysis. RESULTS: HBO treatment recovered SCI-induced descent of BBB scores on POD 14, (1.25 ± 0.75 vs. 1.03 ± 0.66, P < 0.05), 21 (5.27 ± 0.89 vs. 2.56 ± 1.24, P < 0.05), and 28 (11.35 ± 0.56 vs. 4.23 ± 1.20, P < 0.05) compared with the SCI group. Significant differences were found in the mRNA levels of SDF-1 (mRNA: day 21, SCI + HBO vs. SCI + HBO + AMD, 2.89 ± 1.60 vs. 1.56 ± 0.98, P < 0.05), CXCR4 (mRNA: day 7, SCI + HBO vs. SCI, 2.99 ± 1.60 vs.1.31 ± 0.98, P < 0.05; day 14, SCI + HBO vs. SCI + HBO + AMD, 4.18 ± 1.60 vs. 0.80 ± 0.34, P < 0.05; day 21, SCI + HBO vs. SCI, 2.10 ± 1.01 vs.1.15 ± 0.03, P < 0.05), and BDNF (mRNA: day 7, SCI + HBO vs. SCI, 3.04 ± 0.41 vs. 2.75 ± 0.31, P < 0.05; day 14, SCI + HBO vs. SCI, 3.88 ± 1.59 vs. 1.11 ± 0.40, P < 0.05), indicating the involvement of SDF-1/CXCR4 axis in the protective effect of HBO. CONCLUSIONS HBO might promote the recovery of neurologic function after SCI in rats via activating the SDF-1/CXCR4 axis and promoting BDNF expression.
Animals ; Blotting, Western ; Brain-Derived Neurotrophic Factor ; metabolism ; Disease Models, Animal ; Hyperbaric Oxygenation ; methods ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, CXCR4 ; metabolism ; Receptors, Interleukin-8A ; metabolism ; Spinal Cord Injuries ; metabolism ; therapy

OBJECTIVETo establish a method to determine lead and Isotope Ratios in whole blood by inductively coupled plasma mass spectrometry (ICP-MS).

METHODSThe whole blood samples was removed protein by 5% nitric acid , Online join thallium (Tl) as internal standard substance, used lyophilized bovine blood lead and cadmium standard material (GBW09139h and GBW09140h) for quality control of blood lead concentrations. Lead isotope standard substances (GBW04426) is used to determine the correction factor, lead isotope ratios will lead isotope standard reference material NIST SRM981 by icp-ms with samples for testing.

RESULTSOptimize the detection method, detection of blood lead and lead isotope, and the method of linear range r >0.9999, GBW09139h and GBW09140h test results are within the scope of quality control. NIST SRM981 isotope determination precision RSD<1%, NIST SRM981 test results and the certificate of value close to.

CONCLUSIONThe method is simple and convenient data is reliable, can meet the total lead (pb) in blood and former isotope simultaneous determination.

Animals ; Cattle ; Lead ; analysis ; blood ; Lead Radioisotopes ; analysis ; blood ; Mass Spectrometry ; methods