Alveolar Epithelial Cells ; cytology ; metabolism ; Animals ; Apoptosis ; Lung ; drug effects ; metabolism ; Nitric Oxide ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Tyrosine ; analogs & derivatives ; metabolism

AIM: To assess the impact of diabetes on corneal endothelial cells through the quantitative analysis of corneal endothelial cell morphology for patients with diabetics.
METHODS: The corneal thickness and endothelial cell morphology of 360 eyes of 299 cases were detected using full automatic corneal endothelial cell analyzer. The normal control group included 175 eyes of 148 cases, and there were 185 eyes of 151 cases for the patients with diabetes, 110 eyes of 92 cases for the non-proliferating phase group and 75 eyes of 59 cases for the proliferating phase group. The average density of central corneal endothelial cells, proportion of hexagonal cells, coefficient of variation and corneal thickness were compared among groups, and then the statistical analysis was conducted.
RESULTS: Compared with the cornea of the normal group, in the diabetes group, the coefficient of variation of corneal endothelial cells and central corneal thickness increased, while the average density of central corneal endothelial cells and proportion of hexagonal cells decreased, showing a significant difference (P<0. 05). Compared with the cornea of non-proliferating phase group, in the proliferating phase group, the density of central corneal endothelial cells decreased, the coefficient of variation of corneal endothelial cells increased, while the proportion of hexagonal cells decreased with a significant difference (P<0. 05), and the central corneal thickness increased, showing no significant difference(P>0. 05).
CONCLUSION: Compared with the cornea of normal control group, in the diabetes group, the corneal endothelial cells show abnormal morphology, which aggravates with the severity of lesions, especially for the significant changes in the coefficient of variation and the proportion of hexagonal cells. As a result, the corneal resistance to damage in patients with diabetes will decrease.

Objective To study the effects of the recipe for activating blood circulation and nourishing qi on expression of mitochondrial ribosomal protein L51 (MRPL51) in CVB3 infection model in rat cardiac myocytes, to reveal the pathogenesis of CVB3 myocarditis in the genetic level, to explore the therapeutic mechanism of the recipe for activating blood circulation and nourishing qi on CVB3 myocarditis, and to confirm the validity of the recipe for activating blood circulation and nourishing qi on CVB3 myocarditis. Methods After establishing CVB3 infection model and treatment model with recipe for activating blood circulation and nourishing qi by culturing neonatal rat myocardial cells, a modified suppression subtractive hybridization (SSH) was used to isolate differentially expressed genes between two model groups. These results were further verified by fluorescence RT-PCR. Results The results of SSH showed that gene expression of the treatment group was lower than that of the CVB3 infection group. The results of fluorescent RT-PCR which agreed with that of SSH displayed the threshold cycle number (Ct) in the treatment group was higher than the virus group. Conclusion Up-regulation of MRPL51 might be one of the pathogenesis of CVB3 myocarditis. The recipe for activating blood circulation and nourishing qi could treat viral myocarditis by regulating the expression of MRPL51.

Objective To investigate the role of RANKL/RANK/OPG system in bone metabolism of ankylosing spondylitis (AS) by detecting bone mineral density, bone metabolism factors such as osteoprotegerin (OPG), soluble receptor activator of nuclear factors-κB ligand (sRANKL) and the expression of membrane-bound (mb) RANKL in the peripheral blood T lymphocytes. Methods Bone mineral density of AS patients were measured by dual-energy X-ray absorptiometry (DEXA) and serum levels of OPG, sRANKL,tartrate resistant acid phosphatase 5b (TRACP-5b) and bone alkaline phosphatase (BALP) were determined by enzyme-linked immunosorbent assay (ELISA). The percentages of CD4+/RANKL+ and CD8+/RANKL+ in the peripheral blood were detected with flow cytometry. T-test, x2-test were used for statisical analysis. Results ① The incidence of osteopenia and osteoporosis in AS was 47% and 37% respectively. ② Serum RANKL,TRACP-5b levels and RANKL/OPG ratio were higher in AS patients than those in normal controls (P<0.05).But there was no significant difference in OPG and BALP between AS patients and normal controls. ③There were positive linear correlation between serum levels of RANKL and OPG, sRANKL and TRACP-5b, OPG and TRACP-5b in AS (P<0.01). ④ The prevalence of CD4+/RANKL+ cells in the peripheral blood of AS patients was significantly higher than that in the normal controls (P<0.05). Conclusion There is a high incidence of bone loss in AS patients. Increased bone resorbtion is the feature of bone metabolism in AS.RANKL/RANK/OPG system may play an important role. The imbalance of RANKL/RANK/OPG system may be one of the bone loss mechanisms of AS. CD4 + T lymphocyte may play an important role in osteoclasts differentiation and bone resorption in AS by up-regulating the expression of RANKL.

Informed consent right is not just for basic ethical consideration, but is important for protecting patient's right by law, which is expressed through informed consent contract. The appraised individuals of forensic clinical examination have the similar legal status as the patients in medical system. However, the law does not require informed consent right for the appraised individuals. I recommend giving certain informed consent right to the appraised individuals in the forensic clinical examination. Under the contracted relationship with the institution, the appraised individuals could participate in the examination process, know the necessary information, and make a selected consent on the examination results, which can assure the justice and fairness of judicial examination procedure.
Forensic Medicine ; Humans ; Informed Consent ; Patient Participation

Abstract: Objective To investigate the influence of the variation of SARS-CoV-2 on the clinical feature, and to provide early warning signs for the variation of SARS-CoV-2 in clinical work. Methods From Jan 2, 2021 to Jun 30, 2021, a total of 105 COVID-19 patients were included in the study using a case-control method. Nasal swab samples were collected from the study subjects, the viral genes were sequenced, and patients were divided into Delta variant group and non-Delta variant group according to their gene sequences. Clinically relevant data were collected from the two groups, and indicators such as days of hospitalization, age distribution, lymphocytes, neutrophils, B lymphocytes, NK cells, IL-4, and IL-10 were compared; subgroup analysis was performed based on the number of days of viral negativity in the study subjects as the basis for grouping, and differences in immunological characteristics were compared, including lymphocytes, neutrophils, B lymphocytes, NK cells, IL-4, IL-10, etc. Results The theoretical hospitalization days of Delta variant group were (22.2±8.33) d, which were significantly longer than (17.6±10.50) d of non-Delta variant group (t=2.396, P<0.05). The total lymphocyte count and IL-4 of Delta variant group were (1.22±0.86) ×109/L and (0.80±0.23) ng/mL, which were significantly lower than corresponding (1.91±0.70) ×109/L and (1.59±0.59) ng/mL of non-Delta variant group (t=4.329, 9.072, P<0.05), while IL-10 was (7.16±7.77) ng/mL, which was significantly higher than (4.26±3.91) ng/mL of non-Delta mutation group (t=1.980, P<0.05). Subgroup analysis showed that the total lymphocyte count and IL-4 concentration in Delta variant group were (1.04±0.60) ×109/L and (0.74±0.25) ng/ml, which were significantly lower than corresponding (1.62±0.56) ×109/L and (1.56±0.52) ng/mL in non-Delta variant group, in patients with delayed discharge (P<0.05). Conclutions SARS-CoV-2 variant has an impact on clinical manifestations. The patient's B cell count and IL-10 concentration increased or IL-2 and IL-4 concentration decreased within 12 hours of admission indicated variant virus infection. The decrease of total lymphocyte count, especially T lymphocyte reduction, strongly suggests discharge delay due to viral clearance disorder.

NOD mice were treated intraperitoneally with tripterygium wilfordii ployglycosidium (TWP) for 4 weeks to observe the incidence of cyclophosphamide accelerated diabetes.The apoptosis of?-cells was detected by TUNEL,the expression of caspase-3 in islets of the NOD mice was detected by immunohistochemistry and the expression of caspase-8 mRNA in pancreas by RT-PCR.The results revealed that the incidence of diabetes in TWP group was lower than that in control group.The apoptosis index of?-cells was decreased in TWP group. The expression of caspase-3 in islets and the expression of caspase-8 mRNA in pancreas in TWP group were lower than those in control group.

Child ; Humans ; Sleep Wake Disorders ; diagnosis ; etiology ; Tourette Syndrome ; complications

Background Choroidal neovascularization (CNV) is a main cause of visual impairment in many retinal diseases.To create an ideal CNV animal model is very important for the experimental and clinical study of CNV.The assessment method of repeatable and reliable for CNV model is still seldom.Objective This experiment was to explore the label value of fluorescent antibody for visualizing and quantifying the morphologic changes associated with laser-induced CNV.Methods Laser-induced CNV models were created in 30 eyes of 15 male SPF C57BL/6J mice by Krypton red laser irradiating fundus 2 spots around the optical disc with the wavelength 647.1nm,power 260 mW,spot diameter 50μm and exposure time 0.05 seconds.The CNV was evaluated at 5 minutes,4,7,14 and 28 days after laser injury by using fundus photography and fundus fluorescein angiography (FFA),and the successful models were identified as the rupture of Bruch's membrane.The mice were then immediately sacrificed and the eyeballs were enucleated to prepare the choroidal flatmounts.The posterior eye cups were fluorescently labeled with markers of cell nuclei (DAPI,4',6'-diamino-2-phenylindole),endothelial cells (isolectin-B4),and filamentous actin (phalloidin).The CNV areas from specimens were measured by Image pro plus 6.0.Two eyes from one matched mouse without receiving photocoagulation were used as the controlls.This study followed the Standard of Association for Research in Vision and Ophthalmology.Results No any CNV was seen in photocoagulated eyes in 5 minutes and 4 days after laser irradiation.The first sign of CNV appeared at 7 days following photocoagulation.The incidence of fluorescein leakage was 76.47% (26/34),81.81% (18/22),50.00% (5/10) at 7,14 and 28 days,respectively.The fluomicroscope examination showed that in unphotocoagulated areas,retinal pigment epithelial (RPE) cells were visualized with a uniform hexagonal array.Immediately after laser exposure,a circular area devoid of fluorescent labeling was observed,indicating disruption of the choroid-Bruch membrane-RPE complex.On the fourth day,cellular debris and fragmented nuclei were presented and an autofluorescent ring was visible at the site of Bruch's membrane disruption.The number of CNV vessels increased exponentially during the next 3 days.At 7 days,a well-defined isolectin-B4 labeled CNV network was exhibited and lasted for 28 days.The CNV areas were (7.99±0.42)×103μm2,(16.89±3.77)×103μm2,(14.37±4.02)×103μm2 at 7,14 and 48 days after photocoagulation respectively,showing a significant difference among these three groups (F=17.340,P=0.000),and the CNV area was significantly increased in the photocoagulating eyes in 14 days and 28 days compared with 7 days (q=16.46,q=15.54,P＜0.01).Conclusion Fluorescent antibody labeling allows the well identification and measurement of laser-induced CNV lesions in mouse choroid/RPE flatmounts.This technique offers excellent morphologic detail and facilitates the study of critical early events in CNV.CNV complexes are labeled at an early stage,providing a more accurate preclinical evaluation of antiangiogenic molecule.

Objective To study the effects of nuclear factor-?B(NF-?B)decoy oligodeoxynucleotide(ODN)on the preeclamptic umbilical serum induced expression of precollagen Ⅰ,Ⅲ mRNA and tumor necrosis factor-?(TNF-?)in cultured human umbilical artery smooth muscle cells (HUASMC).Methods Primary cultured HUASMC of normal pregnancy were divided into four groups: group A(HUASMC were incubated with umbilical serum of normal pregnancy);group B(HUASMC were incubated with umbilical serum of preeclampsia);group C(HUASMC were transfected with NF-?B cis decoy ODN 48 h before incubation with umbilical serum of preeclampsia);group D(HUASMC were transfected with NF-?B scramble ODN 24 h before incubation with umbilical serum of preeclampsia).NF-?B cis decoy ODN and NF-?B scramble ODN were transfected with cationic lipofectamine to the latter two groups,respectively.The proliferation of human umbilical artery smooth muscle cells was evaluated by methyl thiazolyl tetrazolium and the apoptosis was analyzed by flow cytometry.The expression levels of precollagen Ⅰ,Ⅲ mRNA were detected by RT-PCR,the expression levels of TNF-? were detected by western blot.Results(1)The proliferation of group B(0.19?0.02)and group D(0.18?0.03)was significantly increased as compared with those of group A(0.11?0.02)and group C(0.14?20.02)(P0.05).(5)The expression of TNF-? of group B(0.74?0.11),group C(0.36?0.09)and group D(0.79?0.12)were significantly higher than that of group A(0.15?0.03)(P0.05).Conclusions NF-?B cis decoy ODN could down-regulate the proliferation,as well as the expression levels of precollagen and TNF-? of HUASMC induced by umbilical serum of preeclampsia.NF-?B may play an important role in the pathogenesis of placental artery abnormalities in preeclampsia.