AIM: To observe the clinical effect of combined cataract and primary open-angle glaucoma underwent phacoemulsification and glaucoma drainage device implantation. ·METHODS: We selected in our hospital from January 2014 to February 2016, 42 cases (42 eyes) with primary open-angle glaucoma with cataract. According to random number table method, all of the patients were randomly divided into two groups, control group and study group. In control group, 21 patients ( 21 eyes ) underwent trabecular resection combined phacoemulsification;in study group patients, 21 patients ( 21 eyes ) , underwent EX-PRESS glaucoma drainage device combined phacoemulsification. Compared parameters included postoperative complications and filtering bleb, visual acuity, intraocular pressure ( IOP ) and other clinical indicators between two groups. ·RESULTS: Preoperative IOP of two groups was no significantly different (P>0. 05). Postoperative IOP at each time point was significantly lower than before treatment (P<0. 05). At 1d, 1 and 4wk after treatment, IOP of the study group was significantly lower than the control group (all P<0. 05); at 12wk after treatment, IOP of the two groups was not significantly different ( P> 0. 05). At 12wk after treatment, surgical success rate of study group was 95%, significantly higher than that of control group 71% (P<0. 05). The postoperative best corrected visual acuity of two groups was no significantly different (P>0. 05). At 12wk after treatment, 21 patients in study group were shown as functional filtering bleb, while in the control group 18 cases was functional filtering bleb. ·CONCLUSION:Using EX-PRESS glaucoma drainage device combined with phacoemulsification in treating cataract with primary open-angle glaucoma is reliable, the curative effect is better than that by trabeculectomy combined with phacoemulsification treatment.

Objective To exclude false positivities in detection of IgM antibodies against hepatitis E vires of genotype 4 (HEV-4) using a truncated immunodominant polypeptide of HEV open reading frames (ORF2). Methods The recombinant ORF2 immunodominant polypeptide corresponding to amino acids (AA) 459-607 and a truncated polypeptide corresponding to AA 472-607 were separately applied to coat ELISA plates. Anti-HEV IgM from 35 serum samples with HEV RNA positive, 69 serum samples from healthy individuals and 117 clinically suspicious HEV RNA positive serum samples was detected by an indirect ELISA and was confirmed by western blot in protein level and RT-PCR detecting in RNA level. Results Western blot analysis showed that the sera from HEV patients reacted with the dimmer of peptide 459-607, but they didn't react with the monomer and peptide 472-607. The ELISA showed that all 35 serum HEV RNA positive samples reacted with peptide 459-607 but not with peptide 472-607 and none of the 69 serum samples from healthy individuals reacted with either polypeptide. Among 117 chnically suspicious HEV RNA serum samples, 5 samples reacted simultaneously with both polypeptides. But the difference between 450 nm absorbance (A450) value was less than 0. 5. Western blot analysis demonstrated that all the 5 serum samples were anti-HEV IgM- negative. The 5 serum samples was detected negative by RT-PCR, indicating that the false pesitivities were caused by non-specific absorption. Conclusions ORF2 peptide 459-607 may be used to detect anti-HEV lgM efficiently. The false positivities caused by non-specific absorption can be largely excluded according to the difference between 45Ohm absorbance (A450) value when serum reacts with both polypeptides.

OBJECTIVETo investigate the differences between clinicopathological features and prognosis of alpha-fetoprotein (AFP) negative (AFP < 20 ng/ml) and positive (AFP ≥ 20 ng/ml) hepatocellular carcinoma (HCC) patients.

METHODSClinicopathological data of 142 AFP-negative and 109 AFP-positive HCC patients who underwent RO radical hepatectomy in the Cancer Hospital of Chinese Academy of Medical Sciences between January 2006 and December 2011 were retrospectively reviewed and analyzed in this study.

RESULTSCompared with the AFP-negative patients, a higher female to male sex ratio, the later Barcelona Clinic Liver Cancer ( BCLC) stage, more liver capsule invasion and poorer Edmondson-Steiner grade were in the AFP-positive cases (P < 0.05 for all). Furthermore, the 1-, 3-, and 5- year overall survival rates were 94.4%, 82.4% and 61.0% in the AFP-negative group and 87.2%, 61.1% and 40.2%, respectively, in the AFP-positive group (P < 0.001). The multivariate analysis with Cox's proportional hazards model showed that AFP status, tumor size and Edmondson-Steiner grade are independent risk factors for survival of all the patients (P < 0.05) , and large tumor and Edmondson-Steiner grades III/IV are independent risk factors for worse survival in AFP-negative patients (P < 0.05). However, large tumor diameter was proved to be an independent risk factor leading to poor prognosis of AFP-positive cases (P < 0.05).

CONCLUSIONHigh levels of AFP indicate that the tumors are more malignant and with unfavorable prognosis.

Asian Continental Ancestry Group ; Carcinoma, Hepatocellular ; chemistry ; mortality ; pathology ; surgery ; Female ; Hepatectomy ; Humans ; Liver Neoplasms ; chemistry ; mortality ; pathology ; surgery ; Male ; Multivariate Analysis ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Risk Factors ; Survival Rate ; alpha-Fetoproteins ; analysis

To explore a new method to identify Moutan Cortex to guarantee its safe use, internal transcribed spacer 2 (ITS2) sequence was used to identify Moutan Cortex and its adulterants. DNA was extracted and target fragments were amplified. Sequences were analyzed and assembled by CodonCode Aligner V3.7.1. Genetic distances were computed and phylogenetic tree was constructed based on kimura 2-parameter (K2P) model by MEGA 5.0. The length of the 20 ITS2 sequences of Moutan Cortex from nine different places is 227 bp, and no variation site was detected. The maximum inter-specificK2P distance of Moutan Cortex is 0, the minimum intra-specific K2P distance is 0.041, the average intra-specific K2P distance is 0.222. According to NJ analysis, Moutan Cortex from different places can get together as one branch with bootstrap support values 99%, which indicates Moutan Cortex can be easily distinguished from its adulterants. Using ITS2 sequence can accurately identify Moutan Cortex and its adulterants, it is an effective supplementary to traditional identification methods.
Base Sequence ; China ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Paeonia ; classification ; genetics ; Phylogeny ; Quality Control

Objective:To study the relationship between somatization symptoms and body mass index (BMI), sleep and cognitive function in patients with depression.Methods:A total of 119 patients with depression were selected from January to December in 2019.According to the score of patient health questionnaire-15(PHQ15), they were divided into mild somatization group ( n=75) and moderate severe somatization group ( n=44). Hamilton depression scale-24(HAMD-24), patient health questionnaire-15, Pittsburgh sleep quality index(PSQI) and Montreal cognitive assessment(MoCA) were used to evaluate all subjects.SPSS 23.0 software was used for data analysis.Independent sample t-test was used to compare BMI, sleep and cognitive function scores between the two groups.Pearson correlation analysis was used to study the correlation between somatization symptoms and sleep quality and cognitive function. Results:There were significant differences in BMI((21.70±3.09)kg/m 2, (23.31±3.51)kg/m 2), PSQI((12.56±4.37), (14.37±3.72)), sleep quality(1.87±0.86), (2.21±0.80)), sleep disorder ((1.24±0.59), (1.65±0.53))and daytime dysfunction((2.45±0.81), (2.77±0.48)) between the two groups ( t=-3.783--2.133, all P<0.05), but no difference was found in cognition ( P>0.05). Correlation analysis showed that after controlling HAMD, PHQ-15 was positively correlated with PSQI, sleep quality, sleep disorder, daytime dysfunction and language score in MoCA ( r=0.205-0.298, all P<0.05). Conclusion:The severity of somatization in patients with depression is related to BMI, sleep quality, sleep disorder, daytime dysfunction and language function, suggesting that they may play an important role in the pathogenesis of depression with somatization.

OBJECTIVETo establish a long-term in vitro culture of the fibroblasts obtained from burn wounds.

METHODSSkin samples were harvested from normal volunteers and the deep partial thickness burn wound in burn patients on the 5th, 10th, 21st, 28th and 35th postburn days (PBDs). The non-dermal tissue was removed from the samples and primed by chlorhexidine solution in concentration of 2.5 g/L. The skin sample was then digested by trypsin-EDTA in concentration of 1.25 g/L and was centrifuged before the cells were harvested and cultured. When the cells grew nearly to form sheet, multiple passage culture, freezing storage and revivification were carried out with routine methods. The cell morphology was continuously observed during the culture. And the cell doubling time was calculated.

RESULTSThe wound-origin fibroblasts exhibited higher purity and better activity. The cellular growth features and gross morphology kept stable during primary and secondary culture, and during freezing storage and after revivification. The cells kept their activity above 80% of their original after many times of revivification.

CONCLUSIONThe establishment of the in vitro culture of fibroblasts from burn wounds might be useful in the exploration of the pathogenesis and therapeutic measures of scars.

Burns ; metabolism ; pathology ; Cell Culture Techniques ; methods ; Cell Division ; Cell Survival ; Cells, Cultured ; Cryopreservation ; Factor VIII ; analysis ; Fibroblasts ; chemistry ; cytology ; Humans ; Immunohistochemistry ; Time Factors

OBJECTIVETo explore the in vitro methods of isolation and culture of human fetal epidermal stem cells (HFESCs) and the feasibility of the cultured cells as the target cells for gene transfection.

METHODSThe HFESCs were isolated by means of type IV collagen rapid adhering method. The culture medium for HFESCs was prepared according to that for human fetal fibroblasts. The cultured cells were identified by immunohistochemistry staining of keratin-19 and integrin-beta1, cell cycle analysis and clone forming rate determination. Then the cultured cells were gene transfected in vitro by liposome mediating method in which eukaryon expression vector pcDNA3.1/VEGF165 containing vascular endothelial growth factor 165 (VEGF165) were transfected into cultured cells, or by virus vector mediating method in which recombinant adenovirus accompanied vector (raav) containing green fluorescent protein (GFP) (raav/GFP) were transfected into the cultured cells, respectively. The results of in vitro gene transfection of HFESCs were observed by immunohistochemisty staining and fluorescence microscope.

RESULTSHFESCs grew well and formed large clones with higher cloning efficiency and higher ratio of G1 cells than keratinocytes. The cultured cells were strongly positive with immunohistochemistry staining of keratin-19 and integrin-beta1. After being gene-transfected by pcDNA3.1/VEGF165, the VEGF165 of HFESCs showed positive immunohistochemistry staining property, while the HFESCs transfected by raav/GFP exhibited strong fluorescence.

CONCLUSIONHFESCs could be isolated and cultured in vitro by means of rapid adherence to type IV collagen. It seemed feasible that HFESCs were gene transfected with liposome or adeno-associated virus as the vector.

Cell Adhesion ; Cell Cycle ; physiology ; Cells, Cultured ; Endothelial Growth Factors ; genetics ; metabolism ; Epidermis ; Fetus ; G1 Phase ; Green Fluorescent Proteins ; Humans ; Immunohistochemistry ; Integrin beta1 ; analysis ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Keratinocytes ; cytology ; Keratins ; analysis ; Luminescent Proteins ; genetics ; metabolism ; Lymphokines ; genetics ; metabolism ; Microscopy, Fluorescence ; Plasmids ; genetics ; Stem Cells ; chemistry ; cytology ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

OBJECTIVETo investigate the therapeutic effect of nanometer silver impregnated dressing on gunshot wounds after being immersed in brine and tapwater in rabbits.

METHODSRabbits were randomly divided into two groups after receiving gunshot wounds in both lower limbs. In group 1, the wounded limbs on the experimental side were immersed in brine for 5 h; in group 2, the wounded limbs on experimental side were immersed in tapwater for 5 h. All the wounds were treated with nanometer silver impregnated dressing on the experimental sides, while those of the control sides were treated with vaseline dressing. Biopsy was done after 30 min and 1, 2, 3, 4, 5 h, respectively.

RESULTSIn group 1, the onset of inflammation around the wounds of the experimental sides was delayed, the inflammatory response was less serious, and the wounds were dry with less exudation compared to the controls. The mean healing time of the entry wounds on experimental and control sides was (29.4 +/- 6.6) d and (36.3 +/- 6.0) d (P < 0.01), respectively, and that of the exit wounds on experimental and control sides was (20.1 +/- 6.0) d and (27.3 +/- 5.7) d (P < 0.01), respectively. In group 2, only one of the experimental wounds showed mild inflammation, while all of the control wounds showed serious inflammation with much exudation. The mean healing time of the entry wounds on experimentsides was (13.0 +/- 1.52) d, while that on control sides was (16.0 +/- 3.10) d (P < 0.01). The mean healing time of exit wounds on experimental sides was (11.0 +/- 2.75) d, and those of the control sides was (15.6 +/- 2.85) d (P < 0.01).

CONCLUSIONThe nanometer silver impregnated dressing can control infection and accelerate healing in gunshot wounds in rabbits.

Animals ; Bacterial Infections ; etiology ; prevention & control ; Bandages ; Female ; Immersion ; Male ; Models, Animal ; Nanotechnology ; Rabbits ; Random Allocation ; Salts ; adverse effects ; Seawater ; adverse effects ; Silver ; pharmacology ; therapeutic use ; Treatment Outcome ; Water ; adverse effects ; Wound Healing ; drug effects ; Wounds, Gunshot ; microbiology ; pathology ; therapy

OBJECTIVETo investigate the feasibility of fabricating tissue engineering skin with human hair follicle bulge cells (HFBCs) to repair full-thickness skin wound.

METHODSHFBCs and dermal papilla cells (DPCs) isolated from human fetal hair follicles by collagenase digestion were cultured, purified and passaged. PGA-collagen scaffolds as bioengineered dermis were randomly divided into A and B groups. The HFBCs and DPCs (1 : 2) were seeded in scaffolds of group A and the equal amount of DPCs was seeded in scaffolds of group B as control. Then the keratinocyte sheets were seeded onto the surfaces of the scaffolds as bioengineered epidermis. The tissue engineering skins were then transplanted to repair the full-thickness wound on the back of nude mice. The wound healing process was observed and the plant histological changes of the transplanted engineered skin was observed with light microscope on 2, 4, 6 post-operation weeks (POW).

RESULTSThe full-thickness defect of nude mice in A and B groups could be effectively repaired by bioengineered skins. On 2 POW, integral epidermal and dermal structures were observed in the wounds in A and B groups, with thin epithelial layer and basement membrane. On 4 POW, epithelial layer became thickening and rete pegs formation was observed in basement membrane in A group, but only thickening of epithelial layer was observed in B group. On 6 POW, rete pegs structure was seen to descend and hair-follicle-like structure was formed, while only thickened epithelial layer with flat basement membrane were formed in B group.

CONCLUSIONFrom the composite skin engineered with PGA-collagen hybrid scaffolds and keratinocytes, HFBCs and DPCs could effectively repair the full-thickness skin defect of nude mice. The hair follicle stem cells participate in the process of anatomic repair of wound, and might be able to induce the repair of skin structure and function.

Animals ; Cell Culture Techniques ; Cells, Cultured ; Dermis ; cytology ; Fetus ; cytology ; Hair Follicle ; cytology ; Humans ; Male ; Mice ; Mice, Nude ; Skin Transplantation ; Skin, Artificial ; Tissue Engineering ; methods

OBJECTIVETo develop a rapid and reproducible method for the culture of human fetal hair follicle bulge cells, and observe the plasticity of its differentiation into sebaceous gland in vitro.

METHODSThe bulge cells isolated from fetal human hair follicles by enzymatic digestion (digestion method) and manual microdissection (conventional method) were cultured and passaged respectively, the efficiency and biological features of cells were investigated , the clone forming efficiency was assayed by MTT, and the expression of K19 was further compared by immunocytochemistry (ABC). The morphological change and the expression of EMA of bulge cells were also observed after induction.

RESULTSBy conventional method, 8-10 bulges were harvested in one hour, 40%-50% of their cells were found to adhere to the culture plate after culturing for 48h, and they became confluent after 14 days. In comparison, about 100 bulges were harvested in one hour by digestion method, the adherence efficiency of their cells was 30% after cultivation for 12h and became confluent after 7 days. The cells grew larger with time, with irregular shape and droplets of lipid around the nucleus. The clone forming efficiency of bulge cells cultured by digestion method was (18.2 +/- 2.1) %, which was much higher than that of cells obtained by conventional method[ (12.7 +/- 3.4) %, P < 0.05]. Immunocytochemistry staining showed that positive staining of K19 was observed in most of the bulge cells, with a large amount of brown granules in the cytoplasm.

CONCLUSIONHuman hair follicle bulge cells can be efficiently cultured and multiplied in vitro, and they retained the characteristics of stem cells. And they have the potential to differentiate into sebaceous glands by induction in vitro.

Cell Culture Techniques ; Cell Differentiation ; Cells, Cultured ; Fetal Stem Cells ; cytology ; Hair Follicle ; cytology ; Humans ; Sebaceous Glands ; cytology