Objective To observe the effect of subretinal injection of retinal pigment epithelium (RPE) cells for RPE in mice.Methods A total of 30 postnatal day 7 C57BL/6J mice were randomly divided into normal mice group,OIR model group and OIR model cell transplanted group,10 mice in each group.The OIR model was induced in mice of OIR model group and OIR model cell transplanted group.The RPE cells were subretinal injected into the RPE of mice in OIR model cell transplanted group.At 20 days after the injection,the RPE thickness was evaluated by fluorescence microscope.The expression of RPE65,Bestrophin and zonula occludens-1 (ZO-1) were estimated by Western blot and real-time quantitative PCR (RT-PCR).Results The thickness of RPE in OIR model mice was thinner than that in normal mice;the thickness of RPE in OIR model cell transplantation mice was significantly thicker than that in the OIR model mice.The results of Western blot and RT-PCR indicated that the differences of protein (F=8.597,18.864,25.691) and mRNA expression (F=39.458,11.461,34.796) of RPE65,Bestrophin,ZO-1 were statistically significant between OIR model group and OIR model cell transplanted group (P＜0.05).Conclusions Subretinal injection of RPE cells can promote RPE thickening.RPE65 and Bestrophin protein relative expression levels increased,ZO-1 protein relative expression levels reduced;mRNA expression levels of RPE65,Bestrophin and ZO-1 genes increased.

Objective:To establish a method for the determination of bergeni in Herba Ardisiae Japonicae. Methods: Herba Ardisiae Japonicae was ultrasonic extracted by methanol after crushed and sieved. An untreated fused-silica capillary (75 μm × 48. 5 cm, 40 cm effective length) was used, the running buffer was 25 mmol·L-1 borax in water, the voltage was 15 kV, the temperature was 25℃, the internal standard was cinnamic acid, the ultraviolet wavelength was set at 240nm, and the running time was 10 minute. Results:Bergeni, the internal standard and the impurities were separated by baseline under the above conditions. Bergeni was linear within the range of 3. 356-167. 800μg·ml-1 with the linear equation of Y =0. 034 9X+0. 060 6(r =0. 997 9), the limit of determination was 0. 839 μg·ml-1(S/N = 3), the limit of quantitation was 3. 356 μg·ml-1(S/N = 10), the average recovery was 99. 6%(RSD = 3. 1%, n = 6), and RSDs of intra and inter-day were lower than 5% (n = 3). Conclusion: The method has the advantages of convenience, fastness, accuracy, and low sample and reagent consumption, which can be used to determine bergeni in Herba Ardisiae Japonicae.

ObjectiveTo explore the effect of neoadjuvant combined postoperative therapy on patients with hepatocellular carcinoma after radical operation. Methods41 postoperative cases were divided into 2 groups at random with 20 cases receiving hepatic artery chemotherapy embolization and biotherapy, and 21 cases receiving supporting treatment only.Results The intrahepatic tumor recurrence rate at 1 year and 2 years in treatment group was 10% (2/20) and 30% (6/20), compared with 43% (9/21) and 62% (13/21) in control group (all P

Serum anti-M2 mitochondrial antibody (AMA-M2) was screened with immune colloidal gold technique in 6 008 individuals aged over 60 years who underwent health check-up in Xuhui District of Shanghai,the positive samples were retested with ELISA method.The results showed that 1.31％ (79/6 008) individuals were AMA-M2 positive; the positive rate was 0.82％ (18/2 186) in males and 1.60％ (61/3 822) in females.Eleven subjects with positive AMA-M2 underwent further investigation,8 cases were diagnosed as primary biliary cirrhosis (PBC) and the remaining 3 were highly suspected as PBC.All 11 individuals were abnormal in biochemical tests of liver function and/or clinical symptoms.

The fovea avascular area (FAZ) is an area of the retina surrounded by a continuous capillary plexus that does not have any capillary structure of its own. FAZ is an important region for the formation of fine vision function. The changes of its morphology and surrounding capillary density reflect the degree of macular ischemia, and are closely related to retinal vascular diseases such as diabetic retinopathy, retinal vein occlusion, Coats disease, idiopathic macular telangiectasia, and retinopathy of prematurity. Early observation of FAZ region changes in patients with retinal vascular disease by optical coherence tomography angiography (OCTA) can evaluate the severity and prognosis of the disease. However, the measurement error of FAZ-related data is still a problem that cannot be ignored. At present, OCTA devices of various manufacturers have different methods and algorithms for measuring and analyzing FAZ, which makes it impossible to compare the measured data between different devices. It is believed that with the continuous progress of OCTA related technology, more accurate data of FAZ regional changes can be obtained, which will bring more help to clinical work.

OBJECTIVE: To explore microcosmic information in chronic gastritis dampness syndrome by using serum proteomics of patients with chronic gastritis dampness syndrome. METHODS: Serum proteomics of 18 dampness syndrome cases, 17 non-dampness syndrome cases in chronic gastritis patients and 8 normal controls were analyzed by surface enhanced laser desorption/ionization-time of flight (SELDI-TOF) protein-chip. RESULTS: There was a high expression trend in three ratios of charge (of an electron) to mass (M/S) of 3.2 kD, 6.4 kD and 8.1 kD of protein expression spectrum from patients with chronic gastritis dampness syndrome, and the wave peak value was over 10 units. There was a low expression trend in above-mentioned sites of protein expression spectrum from patients with chronic gastritis non-dampness syndrome and normal group. The wave peak value of patients with chronic gastritis non-dampness syndrome was about or below 10 units, and the wave peak value in normal group was below 5 units. CONCLUSION: There exists a certain significance to explore biologic theory basis of chronic gastritis dampness syndrome by analyzing the serum proteomics.

OBJECTIVETo explore the genetic basis for a pedigree affected with Chediak-Higashi syndrome (CHS).

METHODSClinical data of two CHS patients from the pedigree was collected and analyzed. Targeted next generation sequencing and Sanger sequencing were conducted to detect potential mutation of the LYST gene.

RESULTSBoth patients presented immunodeficiency, oculocutaneous albinism, and acidophilic inclusion body on bone marrow and blood smears. A homozygous c.6077_6078insA (p.Tyr2026Terfs) mutation was detected in the LYST gene in both patients.

CONCLUSIONGenetic testing can play an important role in the diagnosis of CHS.

Chediak-Higashi Syndrome ; genetics ; Female ; Genetic Testing ; Humans ; Infant ; Infant, Newborn ; Mutation ; Pedigree ; Vesicular Transport Proteins ; genetics

OBJECTIVE: To explore the molecular basis for a pedigree affected with coagulation factor V (FV) deficiency. METHODS: Clinical data of the patient and his family members was analyzed. Targeted capture and next-generation sequencing (NGS) and Sanger sequencing were carried out to detect potential variant of the FV gene. RESULTS: The patient presented with jaundice and prolonged prothrombin time (PT) and activated partial thromboplastic time (APTT). V factor activity measured only 0.1% of the normal level, though the patient had no sign of bleeding. A paternal heterozygous variant c.653T>C (p.F218S) and a maternal heterozygous variant c.3642_3643del (p.P1215Rfs*175) were identified in the FV gene of the patient. His elder brother was a heterozygous carrier of the c.653T>C (p.F218S) variant. c.653T>C(p.F218S) was a known pathogenic variant, while the c.3642_3643del (p.P1215Rfs*175) variant was unreported previously. CONCLUSION Mutations of the FV gene probably underlie the hereditary coagulation factor V deficiency in this patient. NGS combined with Sanger sequencing has detected potential variant with efficiency and provided a reliable basis for clinical and prenatal diagnosis for this family.
Aged ; Factor V ; Factor V Deficiency ; genetics ; Genetic Variation ; Heterozygote ; Humans ; Male ; Mutation ; Pedigree ; Phenotype

OBJECTIVE: To analyze the clinical features, biochemical characteristics and molecular pathogenesis of a girl with isovaleric acidemia. METHODS: Clinical features, blood spot amino acid profiles and urinary organic acid profiles of the patient were analyzed. Targeted capture, next generation sequencing and Sanger sequencing were carried out to detect potential variant of the IVD gene. RESULTS: The patient presented with poor weight gain, poor feeding, lethargy, and a "sweaty feet" odor 10 days after birth. Biochemical test suggested hyperammonemia. Blood spot amino acid profiles displayed a dramatic increase in isovalerylcarnitine (C5: 3. 044, reference range 0.04 - 0.4 μmol/L). Organic acid analysis of her urine sample revealed a high level of isovaleric glycine (669. 53, reference range 0 - 0.5). The child was ultimately diagnosed with isovaleric acidemia, and was found to harbor a paternally derived heterozygous variant c.149G>A (p.R50H) and a maternally derived heterozygous variant c.1123G>A (p.G375S) of the IVD gene. Her elder brother was a heterozygous carrier of c.1123G>A (p.G375S) variant. The c.149G>A (p.R50H) was a known pathogenic variant, while the c.1123G>A (p.G375S) variant was previously unreported. CONCLUSION The pathogenesis of the patient was delineated from the perspective of genetics, which has provided a basis for clinical diagnosis, treatment as well as genetic counseling.
Amino Acid Metabolism, Inborn Errors/genetics* ; Child ; Female ; Heterozygote ; Humans ; Isovaleryl-CoA Dehydrogenase/genetics* ; Male ; Mutation

BACKGROUND AND PURPOSE: Whether or not migraine can cause cumulative brain alterations due to frequent migraine-related nociceptive input in patients is largely unclear. The aim of this study was to characterize longitudinal changes in brain activity between repeated observations within a short time interval in a group of female migraine patients, using resting-state functional magnetic resonance imaging. METHODS: Nineteen patients and 20 healthy controls (HC) participated in the study. Regional homogeneity (ReHo) and functional interregional connectivity were assessed to determine the focal and global features of brain dysfunction in migraine. The relationship between changes in headache parameters and longitudinal brain alterations were also investigated. RESULTS: All patients reported that their headache activity increased over time. Abnormal ReHo changes in the patient group relative to the HC were found in the putamen, orbitofrontal cortex, secondary somatosensory cortex, brainstem, and thalamus. Moreover, these brain regions exhibited longitudinal ReHo changes at the 6-week follow-up examination. These headache activity changes were accompanied by disproportionately dysfunctional connectivity in the putamen in the migraine patients, as revealed by functional connectivity analysis, suggesting that the putamen plays an important role in integrating diverse information among other migraine-related brain regions. CONCLUSIONS: The results obtained in this study suggest that progressive brain aberrations in migraine progress as a result of increased headache attacks.
Brain Stem ; Brain* ; Female ; Follow-Up Studies ; Headache ; Humans ; Longitudinal Studies* ; Magnetic Resonance Imaging ; Migraine Disorders* ; Putamen ; Somatosensory Cortex ; Thalamus