Objective To investigate the significance of the expression of serum LDH and β2-MG in predicting survival of NHL patients and there correlations with clinical parameters. Methods The serum levels of LDH and β2-MG of 429 patients with NHL were measured by rate method and immunoturbidimetry.Besides,the clinical reference of the patients such as gender,age,B symptoms,clinical stages,extranodal sites and Karnofsky evaluation were summarized and grouped.All patients' survival status were achieved via phone and letter. All statistical analyses were performed using the SPSS program for Windows (version 16.0).Results The levels of LDH and β2-MG in age,B symptom,the advanced stage,extranodal sites≥2,Karnofsky evaluation were obviously higher than their counterpart and the difference was significant (P＜0.001). The median of LDH and β2-MG lever in age group was 229 U/L,190 U/L, 2.4 mg/L,1.7 mg/L. In B symptom was 260 U/L,192 U/L,2.3 mg/L, 1.7 mg/L. In clinical stage was 252 U/L, 175 U/L, 2.5 mg/L, 1.6 mg/L. In extranodal sites group was 278 U/L,195 U/L,2.4 mg/L,1.8 mg/L.In Karnofsky evaluation group was 250 mg/L,195 mg/L,2.2 mg/L,1.8 mg/L.LDH level of invasive lymphoma was significantly higher than that of indolent ones (median 211U/L,175 U/L,P=0.002),but there were no difference in the 32-MG level (P=0.937). There were no statistical significance about LDH and β2-MG levels in gender and cell type (P＞0.05).Overall survival rates were different between the abnormal LDH group and the normal LDH group (χ2=119.029, P＜0.001).Overall survival rates were different between the elevated β2-MG group and the normal β2-MG group (χ2 =104.733,P＜0.001).Overall survival rates of the abnormal LDH/β2-MG group was significantly lower than that of normal group (x2=192.326,P＜0.001).Multivariate analysis in Cox regression showed that LDH,β2-MG,age,clinical stages, extranodal sites, Karnmofsky evaluation and aggressive were independent prognostic factors.Conclusion The level of serum LDH and β2-MG can be taken as an auxiliary clinical index to evaluate the prognosis of the NHL patients.

To explore the operation methods and indications of the duodenoscopic papillotomy (IEST) with endoscopic nasobiliary drainage (IENBD) for the treatment of duodenal papilla stenosis during the course of common bile duct operation.The clinical data of 219 cases of cholecystolithiasis with choledocholith and the stenosis of papillary underwent endoscopic sphincterotomy (IEST) plus endoscopic nasobiliary drainage (IENBD) in the Second People's Hospital of Chengdu were retrospectively analyzed.It was successful in 198 cases who had the gallbladder and common bile duct stones removed,and endoscopic papillary dissection was performed and the nasobiliary tube was successfully inserted.Nasobiliary drainage was successful in 186 cases (93.9％) of 198 cases.No liquid outflow was observed in nasobiliary drainage in 7 cases (3.5％).Nasal bile duct slipped early in 5 case (2.5％).Primary closure of bile duct incision was completed in 198 cases.It failed in 4 cases (2.0％) who had the bile leakage with primary closure of duct incision.Mild pancreatitis after operation occurred in 3 cases (1.5％).Nose bile duct ligation was performed in 1 case (0.5％).The overall postoperative complication rate was 4.0％ (8/198).IEST + IENBD in open laparotomy was successful in 21 cases.No perforation of intestine and bile duct,bleeding,severe pancreatitis and other complications and death were detected postoperatively in two groups.During the course of laparoscopy and open laparotomy,IEST + IENBD in treating cholecystolithiasis with choledocholith and the stenosis of papillary and primary closure of duct incision after the endoscopic nasobiliary drainage is safe and effective.

Objective:To investigate the clinical characteristics and gene mutation of seven cases of CDKL5 gene related early-onset epileptic encephalopathy diagnosed by next-generation sequencing.Methods:The clinical data of children with early-onset epileptic encephalopathy from February 2018 to December 2019 in the Department of Neurology, Children′s Hospital Affiliated to Zhengzhou University were retrospectively analyzed. The whole exome sequencing method was used to analyze the entire exome of the proband, and seven cases of CDKL5 gene mutation positive were screened out, and Sanger sequencing verification on family members was performed to identify the source and the characteristics of gene mutations were analyzed.Results:Among the seven children diagnosed with CDKL5 gene related early-onset epileptic encephalopathy, the ratio of male to female was 2∶5, and the age of onset was 15 days to five months of birth. The clinical phenotypes all included different degrees of developmental delay and repeated seizures, which were manifested as general seizures, myoclonic seizures, convulsive seizures or focal seizures; the outcome of use of antiepileptic drugs to control seizures was poor, and some applications of ketogenic diet had better effects. CDKL5 gene mutation sites were all denovo mutations, including NM_003159: c.772_776del (p.K258Efs *10) frameshift mutation, NM_003159.2 (exon: 9-15) heterozygous deletion, CDKL5 hemizygous deletion, NM_003159: c.268 (exon5) G>T (p.E90 *, 941) and NM_003159: c.2578C>T (p.Q860 *, 171) nonsense mutation, NM_003159: c.211A>G (p.Asn71Asp) and NM_001323289: c.545T>C (p.L182P) missense mutation. Among them, c.772_776del (p.K258Efs *10), c.268 (exon5)G>T and c.2578C>T (p.Q860 *, 171) have not been reported. Conclusions:CDKL5 gene related early-onset epileptic encephalopathy is an early onset epilepsy, which is more common in women, and has different forms of seizures. The early electroencephalogram is characterized as severe abnormal brain discharge, and the disease progresses in various forms. There are no specific changes in head magnetic resonance imaging. Different gene mutation sites may lead to different phenotypes and prognostic differences. Many anti-epileptic treatments are ineffective, and ketogenic diets are effective for some patients.

OBJECTIVETo investigate the effect of Hath1 (human atonal homolog 1) overexpression on greater epithelial ridge (GER) cells from postnatal rat cochlea in vitro.

METHODSGER cells were isolated by using a combinatorial approach of enzymatic digestion and mechanical separation from P1 rat cochlear. The GER cell cultures were infected by adenovirus containing Hath1 and enhanced green fluorescent protein (ad-Hath1-EGFP), while transfecting EGFP(ad-EGFP) was as controls. Immunostaining were performed at different time points after adenovirus infection.

RESULTSSome of the infected GER cells became myosin VIIa-positive following ad-Hath1-EGFP infection. The earliest time point to see induction of hair cell differentiation (hair cell marker expression) by ad-Hathl was 5 days post-infection. In contrast, infection of the GER sheet cultures with ad-EGFP control virus did not show any myosin VIIa-positive cells at 3-12 days post-infection in all cultures examined.

CONCLUSIONSGER cells may potentially serve as hair cell progenitors and they are capable of differentiating hair cell-like cells when forced to express Hath1.

Animals ; Animals, Newborn ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; Cell Differentiation ; Cells, Cultured ; Cochlea ; cytology ; Epithelial Cells ; cytology ; Hair Cells, Auditory ; cytology ; Rats ; Rats, Sprague-Dawley ; Transfection

OBJECTIVETo detect the expression of Math1, Hes1 and Hes5 in greater epithelial ridge (GER) cells of rat cochlear and explore their influence on hair cell differentiation.

METHODSPostnatal day 0 (P0), day 1 (P1) , day 3 (P3) day 4 (P4) and day 5 (P5) rat cochlear were dissected respectively and then pure GER cells were separated by a combinatorial approach of attachment and mechanical separation. The total RNA of GER cells was extracted by Trizol one step method and the expression of Math1, Hes1 and Hes5 in GER cells was detected with reverse transcription polymerase chain reaction.

RESULTSMath1 was expressed in P0 - P5 rat GER cells and Hes1 was expressed only in PO - P3 rat GER cells, while there was no expression of Hes5 in P0 - P5 rat GER cells.

CONCLUSIONSProbably only when the expression of Math1 reaches a certain level can it induce GER cells to differentiate into hair cells. Meanwhile this process might controlled by Hes1 to some extent.

Animals ; Basic Helix-Loop-Helix Transcription Factors ; genetics ; metabolism ; Cell Differentiation ; Cochlea ; cytology ; metabolism ; Epithelial Cells ; metabolism ; Gene Expression Regulation, Developmental ; Hair Cells, Auditory ; cytology ; metabolism ; Homeodomain Proteins ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Transcription Factor HES-1

OBJECTIVETo establish in vitro culture systems of greater epithelial ridge (GER) cells from rat cochlear and to investigate the characterization, growth pattern and ultrastructure of GER cells.

METHODSUsing a combinatorial approach of enzymatic digestion and mechanical separation to allow isolation and culture of GER cells from P1 rat cochleae. The dissociated GER cells were cultured in serum-free or 10% fetal bovine serum DMEM respectively. BrdU, phalloidin, ZO1, calretinin and myosin VIIa immunostaining and scanning electron microscope observation were performed in GER cell cultures.

RESULTSThe dissociated GER cell cultures showed positive to ZO1, phalloidin and BrdU staining, but negative to myosin VIIa and calretinin. They assumed a polygonal morphology which was similar to epithelial cells and grew in islands-like patches in medium containing 10% fetal bovine serum while forming spheres in serum-free medium. The GER cells presented significant ability to proliferate in both conditions. Scanning electron microscope showed that there was microvillus and centre bodies but not hair cell specific stereociliary bundles on the surface of GER cultures.

CONCLUSIONSThe GER cell cultures showed significant ability to proliferate and grew in islands-like patches in medium containing 10% fetal bovine serum while forming spheres in serum-free medium. The dissociated GER cells expressed epithelial cell specific marker but not marker of hair cells.

Animals ; Animals, Newborn ; Cell Culture Techniques ; Cells, Cultured ; Cochlea ; cytology ; Epithelial Cells ; cytology ; Hair Cells, Auditory ; cytology ; Rats ; Rats, Sprague-Dawley

The purpose of this study was to transplant neonatal rat with human cord blood Lin(-) cells to test the possibility of this xenograft model. The Lin(-) cells were purified from human cord blood (CB) using negative selection strategy based on different lineage-specific antigens. The Lin(-) cells were injected into the liver of neonatal rats using a microinjector at an average of 5 x 10(5) cells for each. Peripheral blood (PB) and spleen were collected at 2,4 and 8 weeks after injection. Flow cytometry was performed to detect human cells in the rat PB, PCR was used to detect human cells in PB as well as spleen. The results showed that a definite proportion of human cells existed in peripheral blood of chimeric rat and the human specific beta2 microglobulin gene fragments were detected in spleen genomic DNA of chimeric rat. It is concluded that human/rat chimera model can be developed with neonatal rats. Human/rat xenograft model may provide a useful and convenient method for human hematopoietic stem cell assay in vivo.
Animals ; Animals, Newborn ; Cord Blood Stem Cell Transplantation ; DNA ; genetics ; Flow Cytometry ; Humans ; Leukocyte Common Antigens ; blood ; Polymerase Chain Reaction ; Rats ; Rats, Sprague-Dawley ; Spleen ; metabolism ; Transplantation Chimera ; blood ; genetics ; immunology ; Transplantation, Heterologous ; beta 2-Microglobulin ; genetics

Objective:To evaluate the value of indirect immunofluorescence on salt-split skin (IIF-SSS) in the diagnosis of bullous pemphigoid (BP) .Methods:A single-center clinical retrospective study was conducted. Totally, 163 patients with newly diagnosed BP were collected from Hospital of Dermatology, Chinese Academy of Medical Sciences from January 2013 to January 2019, so were 404 controls, including 161 with pemphigus, 67 with eczema, 26 with drug eruption, 23 with erythema multiforme, 18 with prurigo nodularis, etc. Blood samples were collected before the treatment, and IIF-SSS, BP180 NC16A enzyme-linked immunosorbent assay (ELISA) and direct immunofluorescence (DIF) assay were performed to evaluate the value of IIF-SSS in the diagnosis of BP. Measurement data were compared by using t test and Mann-Whitney test, and enumeration data were compared by using chi-square test and Fisher′s exact test or McNemar test. Results:The number of cases positive for IIF-SSS, BP180 NC16A ELISA and DIF assay was 160, 153 and 127 respectively in the BP group, and 0, 18 and 26 respectively in the control group. The sensitivities of IIF-SSS, BP180 NC16A ELISA and DIF assay for the diagnosis of BP were 98.15%, 93.86% and 77.91% respectively, and their specificities were 100%, 95.54% and 93.56% respectively. There was strong consistency in the diagnosis of BP between IIF-SSS and DIF (Kappa coefficient= 0.767, P < 0.001) . Conclusion:IIF-SSS has relatively high sensitivity and specificity for the diagnosis of BP, and can serve as a routine method for diagnosing BP.

Digestive system malignant tumor is one of the common malignant tumors in humans,and its high morbidity and low survival rate at advanced stages bring heavy disease burden to patients,families and society.However,current tumor screening technologies are not suitable for screening in large-scale populations and long-term follow-up because of the invasiveness or complexity.Thus,liquid biopsy,which based on biomarkers such as circulating tumor DNA,circulating tumor cells,exosomes and other new biomarkers,has broad prospects for development in tumor screening.Exosome,secreted by living cells,is a type of extracellular vesicle with the lipid bilayer.Compared to other biomarkers,exosome has the advantages of high stability,wide distribution,and high quantity.The various proteins carried by exosome can reflect the characteristics of the origin cells,and exosome has important research value for the early diagnosis of tumors.This article reviews the studies of exosomal proteins as biomarkers for early diagnosis of digestive system malignant tumors in the past five years,and summarizes the characteristics and limitations of the above studies,so as to provide reference for promoting the clinical transformation of exosomal proteins.

OBJECTIVETo compare the difference of vasomotor functions in aortas segments from Wistar rats between 1-hour and 6-hours after exposure of water-soluble components of fine particulate matter (PM2.5).

METHODSAll 30 Wistar rats were assigned to five groups (n=6 for each group) at random: the blank control group, control group for 1-hour and 6-hours, exposure group for 1-hour and 6-hours. The rats were sacrificed 1-hour or 6-hours later and aorta ring segments were mounted on wire myographs.

RESULTS(1) There was no significant difference in vasomotor functions among three control groups (P>0.05). (2) 1-hour or 6-hours after exposure there was a decrease of contraction elicited by 60 mmol/L KCl in contrast to the control group (P<0.05), whereas no significant change between the exposure group for 1-hour and 6-hours (P>0.05). (3) On the level of 10(-5) or 10(-7) mol/L, 1-hour after exposure there was a decrease in endothelium-dependent acetylcholine (ACh) elicited relaxation precontracted by 10(-6) mol/L NE compared with the control group (P<0.01 or P<0.05), on the level from 10(-5) to 10(-7) mol/L there was a decrease compared with the exposure group for 6-hours (P<0.05), whereas no difference between the exposure group for 6-hours and the control group (P>0.05). On the level from 10(-5) to 10(-9) mol/L, 1-hour after exposure there was a decrease in endothelium-independent sodium nitroprusside (SNP) elicited relaxation precontracted by 10(-6) mol/L NE as compared with the control group (P<0.01 or P<0.05) and a decrease on the level of 10(-6) or 10(-9) mol/L compared with the exposure group for 6-hours (P<0.05), 6-hours after exposure a decrease was caused as compared with the control group on the level from 10(-5) to 10(-7) mol/L (P<0.01 or P<0.05).

CONCLUSIONSInhibition of contraction and impairment of relaxation in aortas should be caused 1-hour after exposure to water-soluble components of PM2.5 in the air, which is weaken 6-hours after exposure.

Acetylcholine ; blood ; Air Pollutants ; toxicity ; Animals ; Aorta, Thoracic ; Male ; Particulate Matter ; toxicity ; Rats ; Rats, Wistar ; Vasoconstriction ; drug effects ; Vasodilation ; drug effects