Aim To improve the sensitivity of phycobiliprotein immunofluorescence assay(PBPIFA), by introducing into bridged avidin biotin(BRAB) technique,known as BRAB-PBPIFA. Methods Using ELISA as a gold standard, effectiveness of three assays(direct PBPIFA, BRAB-PBPIFA and ELISA)were compared in simultaneous detection of standard antigen and 500 serum samples. Results BRAB improved greatly sensitivity of PBPIFA in detecting antigen of low concentration, and detection rate of weakly positive samples could increase by one times and a half. In the applying, protein-free blocking agent Tween 20 to BRAB-PBPIFA,possessed better blocking effect. Conclusion Combination of BRAB technique with PBPIFA can improve the detcctive sensitivity.

Objective: To compare two methods (microbial assay and radioimmunoassay) for measuring plasma folate concentrations, and to examine the relationship between plasma folate levels, and alcohol consumption, tobacco use and body mass index, and the risk of hyperhomocysteinemia in China. Methods: We used a microtiter plate microbial assay and a radioimmunoassay to measure the folate concentration in 88 plasma samples. After comparing the results of these two methods and fitting a regression line, we examined the geographical, seasonal, and gender differences in folate concentration of plasma collected from 2 422 adults in south and north areas in China, and evaluated the association of plasma folate concentration, with alcohol consumption, cigarette smoking, and body mass index, and with the risk of hyperhomocysteinemia, using the data from the two assays. Results: The data from the two assays had a linear relationship ( r =0.879, P =0.000); the regression was Y =0.683 X +0.308 (where X and Y were nature logarithmic transformations of plasma folate by microbial assay and radioimmunoassay, respectively); however, the mean plasma folate levels by microbial assay were much higher than those obtained by radioimmunoassay. Both data sets showed similar plasma folate distributions among Chinese adults, associations with other risk factors, and the risk of hyperhomocysteinemia. We estimated that 19.9% of the Southerners and 67.1% of the Northerners had plasma folate concentrations by radioimmunoassay lower than the 6.8 nmol/L used to define plasma folate deficiency. Conclusion: There is a linear relationship between plasma folate levels determined by microbial assay and radioimmunoassay, but because of the different levels obtained in the two assays, it is difficult to use the microbial assay results to evaluate folate status at this time. The use of 10.5 nmol/L as a cut off for plasma folate deficiency by microbial assay needs further study.

AIM: To study the effect of Jiunaoning Injection on apoptosis and the changes in cytosolic calcium induced by hypoxia/hypoglycemia in SH-SY5Y human neuroblastoma cells. METHODS: The apoptosis and the changes in cytosolic calcium were induced by hypoxia and hypoglycemia .Apoptosis was measured via flow cytometry(FCM), and cytosolic calcium was determined by Fluo-3 fluorometry. RESULTS: The hypoxia/hypoglycemia induced apoptosis and increased the cytosolic calcium in SH-SY5Y cells. Jiunaoning Injection inhibited the apoptosis of the SH-SY5Y cells and decreased the concentration of cytosolic free calcium. CONCLUSION: Hypoxia/hypoglycemia induced apoptosis and increase in cytosolic free calcium in SH-SY5Y cells, which were significantly inhibited by Jiunaoning Injection.

Objective: To introduce a reconstruction procedure of natural sideburn, with combined expanded retroauricular flap and scalp flap. Methods: A retrospective study was produced in Plastic Surgery Hospital, PUMC, from January 2014 to December 2017. Twenty patients (21 sides) underwent sideburn reconstruction with combined expanded retroauricular flap and scalp flap (double pedicled flap, n=3; single pedicled flap, n=17) were included in this study. There were 12 male (12 sides) and 8 female (9 sides), with the mean age of (23.8±3.2) years. The sideburn defect was caused by burn in 19 patients, and it was resulted from hemangioma in 1 patient. Results: The size of flaps ranges from 8 cm×12 cm to 10 cm×16 cm. Venous congestion at the distal end of the flap occurred in 1 patient, which was cured after dressing change, and the sideburn was not affected. The reconstructed sideburns are natural, symmetric, and without obvious scar. The follow-up time was 3-40 months. Fourteen patients were very satisfied with the reconstructed sideburn, and 6 patients were satisfied. No severe complication was observed during follow-up time. Conclusions The combined retroauricular flap and scalp flap is an alternative method for sideburn reconstruction, which provide natural hair distribution, inconspicuous scars and less complications.

Objective: To explore a surgical method for the reconstruction of naturally dynamic double eyelid by fully reserving orbicularis oculi muscle. Methods: Incisional double eyelid blepharoplasty were performed on 352 patients. Orbicularis oculi muscle in incisional and pretarsal area were completely preserved. Muscle was elevated from plate by resecting deep muscular myolemma and muscles to form pretarsal myocutaneous flap. Thereafter, the orbital septum was completely exposed. The free end of muscle flap was fixed to a higher position of the levator aponeurosis. The orbital septum was repaired at the same time. Results: From June 2009 to December 2017, 352 patients were performed double eyelid blepharoplasty using above technique. They are 337 females and 15 males. Fifty-two patients were followed up for 6-60 months, with the mean time of 27 months. All the incisions were uneventfully healed. The new double eyelid presented smooth appearance, and the so-called " sausage" phenomena were not noticed. There was no concave groove or step-like deformity when eyes closed. Forty-five patients were satisfied with the outcomes. Two patients had shallower supratarsal folds on one side, and 1 patient underwent secondary repair. Two patients complained about asymmetry. Two patients suffered from mild unilateral eyelash eversion (without need of reparation). Conclusions The combined application of deep and superficial myolemma release of the orbicularis oculi muscle, and orbital septum reset, could create a double eyelid with natural and dynamic appearance.

Objective To explore the effect of coptisine on the growth of NCI-H1650cells and to evaluate its potential value in the treatment of human non-small cell lung cancer. Methods MTT method was used to ana-lyze cell proliferation. Protein expressions of Bax/Bcl-2 and cytochrome C in NCI-H1650 cells were detected by-Western blot.Apoptosis was analyzed using flow cytometrywithAnnexin V/PI method.ROS concentration was tested with fluorometry.Results Coptisine could significantly inhibit growth of NCI-H1650 cells in a time-and dose-de-pendent manner.Coptisine induced apoptosis in NCI-H1650 cells by inducing ROS accumulation and the following mitochondria mediated apoptosis which was identified by increased Bax expression,Bcl-2 expression was down-reg-ulated,and cytochrome C moved from mitochondria to cytoplasm.ROS inhibitor(N-acetyl cysteine)treatment dra-matically abrogated coptisine-induced growth inhibition and apoptosis.Conclusions This study suggests that copti-sine can induce ROS irritated-and mitochondria-mediated apoptosis in NCI-H1650 cells.Coptisine has a potential value in the treatment of human non-small cell lung cancer.

Objective:To study the clinical phenotype and molecular genetic characteristics of a Chinese Han family with X-linked retinoschisis (XLRS), and to determine the associated gene variations.Methods:A pedigree investigation was performed.The clinical characteristics and pedigree analysis of a Han Chinese family line with XLRS was conducted in August 2021 at the Xiamen Eye Center Affiliated to Xiamen University.All patients and the carriers underwent comprehensive medical history collection and routine ophthalmological examinations, including visual acuity, non-contact tonometer, slit lamp microscope, direct ophthalmoscope, and optical coherence tomography.The proband and some patients underwent medical optometry, fundus photography or wide-angle fundus photography, and electroretinogram examination.Peripheral venous blood samples were collected from the family members, and whole exome sequencing (WES) analysis was performed on the proband samples.For variants screened by WES, the expanded verification in other patients and normal persons in the family was carried out by Sanger sequencing.Multiple bioinformatic tools were used to analyze the pathogenicity of variants.This study protocol was approved by the Ethics Committee of Xiamen Eye Center of Xiamen University (No.XMYKZX-KY-2021-012). Written informed consent forms were obtained from each subject or guardian of minors.CADD, FATHMM and other bioinformatics tools were used to analyze the pathogenicity of the variation sites.Results:The Han XLRS pedigree consisted of 8 individuals in 3 generations.Out of the 3 cases diagnosed with XLRS based on clinical evaluation, all were male.The mother of the proband was a carrier of related genes.There were 5 persons with normal phenotypes.There was no history of consanguineous marriages within the family, and the disease was shown to be intergenerational, which is consistent with the recessive inheritance of the X chromosome.None of the patients had a history of systemic disease or any other abnormal manifestations.The prevailing feature of ophthalmopathy was poor binocular vision since childhood.The proband and his younger brother had spoke split in the macula, and their grandfather showed atrophy of retinal nerve fibers.Genetic analysis revealed a hemizygous variation c. 214G>C: p.Glu72Gln in the RS1 gene in all the patients in this family.The proband's mother was heterozygous at this site, and all other phenotypically normal family members exhibited wild type at this site.This variant was predicted to be a deleterious variation and likely to cause disease based on bioinformatics analysis. Conclusions:The proband and patients in this Han Chinese family have the known c. 214G>C: p.Glu72Gln hemizygous variation of the RS1 gene and exhibit mild XLRS, which was consistent with the recessive inheritance of X chromosome.

Objective:To explore whether there is Warburg effect in keloid fibroblasts (KFs) , hypertrophic scar fibroblasts (HSFs) and atrophic scar fibroblasts (ASFs).Methods:KFs, HSFs, ASFs and NFs were isolated and cultured, respectively, in vitro. Glucose consumption and lactate production of the cells in each group were tested and compared. The mRNA expression of key glycolytic enzymes was detected, including hexokinase 2 (HK2), fructose-2, 6-biphosphatase 3 (PFKFB3), pyruvate kinase isoform M 2 (PKM2) and lactate dehydrogenase A (LDHA). In addition, the effect of 2-deoxy-D-glucose(2-DG) on the proliferation of KFs and NFs were detected and compared.Results:Glucose consumption and lactate production in KFs were 45.5% and 38.1% higher than those in NFs respectively ( P<0.01), while there was no statistical difference in glucose consumption and lactate production between ASFs and NFs, HSFs and NFs ( P>0.05). The mRNA expression of HK2, PKM2 and LDHA genes in KFs were 4.7, 2.7 and 1.8 times higher than those in NFs ( P<0.05), respectively, while the gene expressions in ASFs and HSFs had no statistical differences from those in NFs ( P>0.05). Under the action of 2-DG, the cell proliferation activity in KF group and NF group were reduced by 37.5% and 27.0%, respectively, indicating that the inhibition effect of glycolysis inhibitor on the proliferation activity of KFs was significantly higher than that of NFs. Conclusions:There is Warburg effect in KFs and there are no similar phenomena in HSFs and ASFs.

The etiology and pathogenesis of keloid are still not clear. It is believed that the formation of keloid is due to the joint action of many factors, during which process the external factor of fibroblast play an important role. Reprogramming of glucose metabolism in keloid fibroblasts is a new research direction. TGF-β 1 and its related TGF-β 1/Smad signalling pathway are considered to be the most closely related to the formation of keloid. This article reviews the research progress on extracellular factors, glucose metabolism and signaling pathways of keloids.

Objective:To explore whether there is Warburg effect in keloid fibroblasts (KFs) , hypertrophic scar fibroblasts (HSFs) and atrophic scar fibroblasts (ASFs).Methods:KFs, HSFs, ASFs and NFs were isolated and cultured, respectively, in vitro. Glucose consumption and lactate production of the cells in each group were tested and compared. The mRNA expression of key glycolytic enzymes was detected, including hexokinase 2 (HK2), fructose-2, 6-biphosphatase 3 (PFKFB3), pyruvate kinase isoform M 2 (PKM2) and lactate dehydrogenase A (LDHA). In addition, the effect of 2-deoxy-D-glucose(2-DG) on the proliferation of KFs and NFs were detected and compared.Results:Glucose consumption and lactate production in KFs were 45.5% and 38.1% higher than those in NFs respectively ( P<0.01), while there was no statistical difference in glucose consumption and lactate production between ASFs and NFs, HSFs and NFs ( P>0.05). The mRNA expression of HK2, PKM2 and LDHA genes in KFs were 4.7, 2.7 and 1.8 times higher than those in NFs ( P<0.05), respectively, while the gene expressions in ASFs and HSFs had no statistical differences from those in NFs ( P>0.05). Under the action of 2-DG, the cell proliferation activity in KF group and NF group were reduced by 37.5% and 27.0%, respectively, indicating that the inhibition effect of glycolysis inhibitor on the proliferation activity of KFs was significantly higher than that of NFs. Conclusions:There is Warburg effect in KFs and there are no similar phenomena in HSFs and ASFs.