Objective:To analyze changes of donor corneal endothelial cell density (ECD) and morphology from eye bank before and after keratolasty and the influencing factors.Methods:An observational case series study was performed.A total of 118 donor corneas, retrieved by the Shandong Province Eye Bank between July 2020 and June 2021 for penetrating keratoplasty (PKP) and endothelial keratoplasty (EK) were included.Among them, 99 corneas (83.90%) were used for PKP, and 19(16.10%) for EK.The basic information of donors and the results of corneal quality tests were analyzed and compared with ECD measured by endothelial microscopy one month after keratolasty.Morphological changes in endothelial cells before and after surgery were observed, and factors influencing corneal ECD and morphology were analyzed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Shandong Eye Hospital (No.SDSYKYY20170319).Written informed consent was obtained from each subject.Results:Among the 99 donor corneas for PKP, there were statistically significant differences in preoperative donor corneal ECD and 1-month postoperative ECD of implant among different age groups ( F=18.136, 5.936; both P<0.01), which were lower in the 31-60-year-old group and the >60-year-old group than in the 0-30-year-old group and higher in the 31-60-year-old group than in the >60-year-old group, with statistically significant differences (all P<0.01).There were statistically significant differences in the preoperative donor corneal ECD among different donor cause of death groups ( F=4.524, P<0.01), which was higher in the traumatic accident group compared to the cardiovascular and cerebrovascular disease group, chronic organ failure group and malignant tumor group (all P<0.01).The preoperative donor ECD in the death-tissue retrieval time ≤6 hours group was (2 577.66±284.63)cells/mm 2, which was higher than (2 372.46±399.75)cells/mm 2 in the death-tissue retrieval time >6 hours group, with a statistically significant difference ( t=2.289, P<0.05).There were statistically significant differences in 1-month postoperative ECD among the preservation-surgery time ≤3 days, 3-6 days, and >6 days groups ( F=6.201, P<0.01), with higher ECD in preservation-surgery time ≤3 days groups than in 3-6 days and >6 days groups (both P<0.01).The preoperative donor corneal ECD applied to EK was significantly higher than that applied to PKP ( t=-2.660, P<0.01).ECD at 1 month after surgery applied to PKP was significantly higher than that applied to EK ( t=4.286, P<0.01).The ECD reduction rate was 7.14% (0.01%, 17.69%) and 31.07% (22.11%, 45.86%) in PKP group and EK group, respectively, with a statistically significant difference ( Z=4.969, P<0.01).The ECD was lower in the group with dark area than in the non-dark area group before PKP, with a statistically significant difference ( t=6.789, P=0.011).There was no significant difference in ECD at 1 month after keratoplasty between the two groups ( t=0.005, P=0.945).Multivariate logistic regression model results showed that preservation-surgery time >6 days and the cause of donor death being malignant tumor were risk factors for the appearance of dark areas in donor corneal endothelium ( OR=9.038, P=0.030; OR=6.577, P=0.018). Conclusions:The older the donor, the lower the ECD.Prolonged preservation-surgery time (>6 days) is the main factor contributing to the decline in ECD after keratolasty.Compared to PKP, there is a higher endothelial cell loss after EK.The tissue preservation-surgery time >6 days and the cause of donor death being malignant tumor are the main risk factors affecting the appearance of dark areas in the donor corneal endothelium.But the presence of physiological dark areas does not significantly influence the ECD after surgery.

Objective:To analyze changes of donor corneal endothelial cell density (ECD) and morphology from eye bank before and after keratolasty and the influencing factors.Methods:An observational case series study was performed.A total of 118 donor corneas, retrieved by the Shandong Province Eye Bank between July 2020 and June 2021 for penetrating keratoplasty (PKP) and endothelial keratoplasty (EK) were included.Among them, 99 corneas (83.90%) were used for PKP, and 19(16.10%) for EK.The basic information of donors and the results of corneal quality tests were analyzed and compared with ECD measured by endothelial microscopy one month after keratolasty.Morphological changes in endothelial cells before and after surgery were observed, and factors influencing corneal ECD and morphology were analyzed.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Shandong Eye Hospital (No.SDSYKYY20170319).Written informed consent was obtained from each subject.Results:Among the 99 donor corneas for PKP, there were statistically significant differences in preoperative donor corneal ECD and 1-month postoperative ECD of implant among different age groups ( F=18.136, 5.936; both P<0.01), which were lower in the 31-60-year-old group and the >60-year-old group than in the 0-30-year-old group and higher in the 31-60-year-old group than in the >60-year-old group, with statistically significant differences (all P<0.01).There were statistically significant differences in the preoperative donor corneal ECD among different donor cause of death groups ( F=4.524, P<0.01), which was higher in the traumatic accident group compared to the cardiovascular and cerebrovascular disease group, chronic organ failure group and malignant tumor group (all P<0.01).The preoperative donor ECD in the death-tissue retrieval time ≤6 hours group was (2 577.66±284.63)cells/mm 2, which was higher than (2 372.46±399.75)cells/mm 2 in the death-tissue retrieval time >6 hours group, with a statistically significant difference ( t=2.289, P<0.05).There were statistically significant differences in 1-month postoperative ECD among the preservation-surgery time ≤3 days, 3-6 days, and >6 days groups ( F=6.201, P<0.01), with higher ECD in preservation-surgery time ≤3 days groups than in 3-6 days and >6 days groups (both P<0.01).The preoperative donor corneal ECD applied to EK was significantly higher than that applied to PKP ( t=-2.660, P<0.01).ECD at 1 month after surgery applied to PKP was significantly higher than that applied to EK ( t=4.286, P<0.01).The ECD reduction rate was 7.14% (0.01%, 17.69%) and 31.07% (22.11%, 45.86%) in PKP group and EK group, respectively, with a statistically significant difference ( Z=4.969, P<0.01).The ECD was lower in the group with dark area than in the non-dark area group before PKP, with a statistically significant difference ( t=6.789, P=0.011).There was no significant difference in ECD at 1 month after keratoplasty between the two groups ( t=0.005, P=0.945).Multivariate logistic regression model results showed that preservation-surgery time >6 days and the cause of donor death being malignant tumor were risk factors for the appearance of dark areas in donor corneal endothelium ( OR=9.038, P=0.030; OR=6.577, P=0.018). Conclusions:The older the donor, the lower the ECD.Prolonged preservation-surgery time (>6 days) is the main factor contributing to the decline in ECD after keratolasty.Compared to PKP, there is a higher endothelial cell loss after EK.The tissue preservation-surgery time >6 days and the cause of donor death being malignant tumor are the main risk factors affecting the appearance of dark areas in the donor corneal endothelium.But the presence of physiological dark areas does not significantly influence the ECD after surgery.

Objective To elucidate the clinical and pathological characteristics of the patients with thymoma-associated glomerulonephropathy.Methods In this retrospective study,the clinicopathologic characteristics of patients diagnosed as thymoma-associated glomerulonephropathy inPeking University First Hospital during the period between Oct 2008 and Jun 2017 were analyzed,including the histological classfication of thymoma,the clinicopathological features and the short-term prognosis.Results Altogether twelve patients were included with an average age of (55+ 16) years;male/female ratio was 3∶ 1.The B2 type thymoma was the most common type.Nine cases also suffered from myasthenia gravis,and eight cases of glomerulopathy accompanied by thymoma activity.The clinical presentation of glomerulopathy included nephrotic syndrome (11/12),acute kidney injury (10/12).Eleven patients received renal biopsy,among which five cases were minimal change nephropathy,three cases were membranous nephropathy,and the other three cases were focal segmental glomerulosclerosis,thrombotic microangiopathy and endocapillary proliferative glomerulonephritis,respectively.Eleven patients received immunosuppression therapy.After a median 12 months follow up,the proteinuria decreased in 7 cases,and renal function completely or partially recovered in 6 cases.Conclusions Minimal change disease is the most frequent pathological type of thymoma-associated glomerulonephropathy.Immunotherapy with glucocorticoid as first-line drug may be considered for thymoma-associated glomerulonephropathy with surgery,chemoradiation contraindications or non-remission of kidney disease after anti-tumor therapy.

Objective: To understand the potential viral pathogens other than enteroviruses existing in samples of hand foot and mouth disease (HFMD) patient and study their molecular feature and genotype. Methods: The deep sequencing analysis of a fecal specimen collected from HFMD patient was conducted by metagenomics and bioinformatics. Results: Enterovirus A71 and sapovirus mixed infection was found in this case. The nucleic acid of sapovirus was confirmed positive by RT-PCR and the 7 429 bp complete genome sequence of sapovirus was obtained by assembling sequencing reads which consisted of 3 open reading frames. Phylogenetic analysis revealed that this strain of virus should belong to the genotype 1 of sapovirus having a homology of 99.4% with sapovirus Hu/G1/Zhejiang1/China/2014 strain, which is a currently predominant genotype circulating in China. Conclusions The sapovirus, which is a predominant strain circulating in China, was a mixed infected causative agent existing in HFMD sample identified by deep sequencing. This study will serve as a reference for pathogen detection of HFMD and diarrheal related diseases, as well as provide a sequence reference for molecular feature study of sapovirus in China in the future.

Objective To explore the application effects of solution focused approach in psychological intervention to the anxiety and depression before operation in lung cancer patients. Methods A total of 92 lung cancer patients were divided into control group and observation group, with each group 46 cases. The patients of control group was given routine psychological nursing, while the patients of observation group received a 5 -steps psychological intervention of solution focused approach on the basis of routine psychological nursing. The patients were elevated for the level of anxiety and depression by the self-rating anxiety scale ( SAS) and the self-rating depression scale ( SDS) on the day of admission and the day before operation, and results were compared between two groups. Results The differences of SAS and SDS scores between two groups were not statistically significant before intervention (P >0. 05). After psychological intervention, the scores of SAS and SDS of observation group were ( 45. 85 ± 7. 49 ) and ( 49. 03 ± 7. 46 ) points respectively, which were lower than (50.62±8.01)and(54.36±6.72)inthecontrolgroup(t=2.950,3.600;P<0.05).Conclusions The application of solution focused approach in preoperative psychological intervention can effectively relieve the anxiety and depression before operation in patients with lung cancer.

Purpose To study the clinicopathological characteristics of primary pancreatic neuroendocrine neoplasms. Method 60 cases of resected pancreatic neuroendocrine neoplasms according to the WHO (2010) classification of the digestive system of neuroen-docrine tumor to evaluate morphological standard, and combining with the literature to discuss the clinicopathological characteristics. Results Among the 60 cases, 23 cases were male patients, the rest were females, with male and female ratio of 1 ∶ 1. 61. The age of the patients were ranged from 19 to 69 years, with mean age of 49. 38 ± 11. 60 years. Tumor maximum diameter ranged from 0. 5 to 16 cm, and the mean diameter was 3. 29 ± 3. 53 cm. 30 cases located in the pancreatic head, 27 cases in the body and end of the pancre-as and 3 cases in the neck. Pathological examination showed the G1 (24 cases), G2 (25 cases), G3 (9 cases), and mixed adenon-euroendocrine carcinoma ( MANEC) in 2 cases. Immunohistochemical staining showed that NSE, CgA, Syn, and CD56 were diffusely positive expression. 45 patients were followed up for 4~80 months, 7 cases died, of which 1 case was G2, 4 cases were G3, and 2 ca-ses were MANEC. Conclusion Primary pancreatic neuroendocrine neoplasms is a relatively rare pancreatic malignant tumor, and the diagnosis is based primarily on histologic features and immunohistochemical examination. Accurate pathological assessment has impor-tant value to guide clinical treatment and prognosis.

Objective To investigate the effect of different loading doses of atorvastatin in patients with stable plaques.Methods Consecutive 174 patients with stable plaque who underwent coronary arteriongraphy (CAG) and intravascular unltrasound(IVUS) were randomly assigned to receive 10 mg atorvastatin treatment (group 10 mg,n =47),20 mg atorvastatin treatment(group 20 mag,n =45),40 mg atorvastatin treatment (group 40 mg,n =43) and 80 mg atorvastatin treatment (group 80 mg,n =39).The endpoints including low density lipoprotein-cholesterol(LDL-C),high density lipoprotein-cholesterol (HDL-C),high-sensitivity C-reactive protein (hs-CRP) levels,necrotic and plaque volumes,were assessed after 3-6 months' follow-up.Results Mean LDL-C,HDL-C,hs-CRP,the percentage of necrotic,plaques volumes were similar at baseline (P ＞ 0.05).During 3-6 months of follow up:(1)LDL-C levels in group 10 mg,20 mg,40 mg,80 mg were lower than at baseline (t =3.12,4.23,3.26 and 5.21 respectively,P ＜ 0.01).There was significant difference between the 20 mg group and the 40 mg group(P ＜ 0.05) ; (2) There was no significant difference of HDL-C levels in 10 mg,20 mg and 40 mg groups after atorvastatin treatment.However,its level was significantly higher in the 80 mg group after atorvastatin treatment than other dose groups(P ＜ 0.05) and were higher than at baseline(t =2.35,P ＜ 0.01) ;(3)the 80 mg group's hs-CRP levels decreased significantly after treatment than at baseline((3.59 + 1.07)mg/L vs (6.10 + 2.12) mg/L,t =2.37,P ＜ 0.01);(4)According to the VH of IVUS,the percentage of necrotic in 10 mg group became higher than at baseline ((16.54 + 1.76) ％ vs.(7.83 + 1.03) ％,t =2.38,P ＜0.01) and conformed to unstable plaques diagnostic criteria(＞ 10％).There was no significant difference in group 20 mg,40 mg and 80 mg with at baseline (t =1.24,0.21,0.69 respectively,P =0.069,0.846,0.643respectively) ; (5)Plaques volumes in group 10 mg,20 mg were not larger than at baseline.However,in group 40mg and 80 mg.Plaques volumes were smaller than at baseline ((30.69 ± 8.12) mm3 vs (37.09 + 12.01)mm3,t=l.29,P=0.019;(24.99±l.01) mm3 vs (36.47+14.68) mm3,t =2.62,P＜0.01).Conclusion The effects of atorvastatin on stable plaques vary with doses.(1) For LDL-C,the use of atorvastation 20 mg/d can make LDL-C reaching standard,and atorvastation 40 mg/d was superior to 20 mg/d and similar to 80 mg/d.(2)For HDL-C,atorvastation 80 mg/d can make it higher.(3)Atorvastation 80 mg/d can make hs-CRP lower.(4)Atorvastation≥20 mg/d can make plaques stable and atorvastation 80 mg/d was superior to 20 mg/d and 40mg/d.Atorvastation 40-80 mg/d can make plaques dwindled.

BACKGROUND: Conventional hemodialysis mainly for cleaning uremic micro molecule substance, such as urea nitrogen or creatinine; however, few hemodialyses can clean uremic middle molecule substances (MMS). With prolonged dialysis duration, MMS accumulates in vivo and induces a series of complications. OBJECTIVE: To compare the efficiency of adsorptive dialysis (hemoperfusion unites hemodialysis) and conventional hemodialysis in cleaning uremic MMS. METHODS: Totally 60 maintenance hemodialysis patients were averagely divided into the adsorptive dialysis group and conventional hemodialysis group. First of all, hemoperfusion apparatus and dialyser were connected in series to take the adsorptive dialysis in the adsorptive dialysis group (hemoperfusion apparatus were equipped before dialyser). 120 minutes later, the hemoperfusion apparatus was toke off and continues to hemodialysis for 120 minutes. Duration of conventional hemodialysis was 240 minutes. Changes in clinical symptoms and levels of liver function, kidney function, serum electrolytes, hemocytes and uremic MMS were observed prior to and after treatment. RESULTS AND CONCLUSION: Adsorptive dialysis could remove the MMS notably. Compared with the conventional hemodialysis group, a single 120 minutes treatment could decrease MMS significantly (P < 0.05). The platelet levels were obviously decreased in the adsorptive dialysis group after treatment (P < 0.05), which were significantly different from the conventional hemodialysis group (P < 0.05). There was no significant difference in liver function, kidney function or serum electrolytes concentration. But related symptoms, such as the skin itch, sleep disorders and myalgia, were relieved more or less.

OBJECTIVETo provide molecular evidences for phylogenetic analysis by studying ITS sequences of Rehmannia glutinosa from different areas.

METHODThe DNAs were extracted from leaves of R. glutinosa by means of CTAB method. The products of PCR amplification were cloned . The data were analyzed by MEGA4.0 software.

RESULTThe results showed that the size of the ITS of R. glutionsa tested was from 613 to 614 bp and the length variation was only 1 bp. The sequence of ITS1 was 224-225 bp, and G + C content varied from 60.4% to 63%. The sequence of ITS2 was 224-225 bp and G + C content varied from 57.1% to 65.3%. The sequence of 5. 8S rDNA was 164 bp, it's very conservative in these species. Phylogram tree based on ITS sequence data indicated that the kinship between Bejing No. 2 R. glutinosa and the others were far. There was obvious diversity within wild R. glutinosa varieties, while there was no different among cultivated R. glutinosa varieties. In cultivated R. glutinosa varieties, there was no diversity between R. glutinosa varieties from Henan and those from others provinces. In wild varieties, R. glutinosa from Shengnongshan and Qingtianhe of Henan province showed a closer systematic relationship with cultivated R. glutinosa from Shandong province, while there was no difference between wild R. glutinosa varieties and cultivated varieties from Henan and Shanxi provinces.

CONCLUSIONThe genetic relationship among R. glutinosa varieties was very close, there was no distinct systematic differentiation.

DNA, Plant ; analysis ; Genetic Variation ; Molecular Sequence Data ; Phylogeny ; Plant Leaves ; genetics ; Polymorphism, Single Nucleotide ; RNA, Ribosomal, 16S ; analysis ; Rehmannia ; classification ; genetics

Heparinase III is an enzyme that specifically cleaves certain sequences of heparan sulfate. Previous reports showed that this enzyme expressed in Escherichia coli was highly prone to aggregation in inclusion bodies and lacks detectable biological activity. In this paper, we fused a glutathione-S-transferase (GST) tag to the N-terminus of heparinase III gene and expressed the fusion protein in Escherichia coli to develop an expression system of soluble heparinase III. As a result, approximately 80% of the fusion protein was soluble. The protein was then purified to near homogeneity via one-step affinity chromatography. A 199.4-fold purification was achieved and the purified enzyme had a specific activity of 101.7 IU/mg protein. This represented 32.3% recovery of the total activity of recombinant GST-heparinase III. The maximum enzyme production was achieved when bacteria were induced with 0.5 mmol/L isopropyl-beta-D-thiogalactoside at 15 degrees C for 12 h. The enzyme showed maximum activity at 30 degrees C and pH 7.5. And the enzyme activity was stimulated by 1 mmol/L Ca2+ and 150 mmol/L NaCl.
Escherichia coli ; genetics ; metabolism ; Flavobacterium ; enzymology ; genetics ; growth & development ; Glutathione Transferase ; biosynthesis ; genetics ; Heparin Lyase ; biosynthesis ; genetics ; isolation & purification ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; isolation & purification