Objective To observe the effects of velcade on inflammatory reaction and cell apoptosis after ischemia-reperfusion injury, and to explore the neuroprotective mechanism of velcade.Methods The 15 male Wistar rats were randomly divided into sham-operated group, physiological saline control group and velcade-treated group (n= 5, each). The model of temporary middle cerebral artery occlusion (MCAO) was applied and reperfused after 2 hours. Immediately after the reperfusion, all rats were performed intraperitoneal injection with velcade 0. 2 mg/kg in velcadetreated group, and with the same volume of physiological saline in control group. After 24 hours, the rats were decapitated in all groups. The apoptosis cells were found by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) and the expressions of nuclear factor-κBp65 (NF-κBp65) and interleukinh-1β (IL-1β) were detected by immunohistochemistry. Results The immunologically positive cells of NF-κBp65, IL-1β and apoptosis cells were occasionally found in shamoperated group [(1.21 ± 0. 16)/400 power, (11.56 ± 0. 99)/400 power and (2. 88 ± 0. 27)/400 power], while a lot of immunologically positive cells of NF-κBp65, IL-1β and apoptosis cells were found in velcade-treated group and control group. The control with compared group, these cells were significantly more in the velcade-treated group [(56.28± 1.95)/400 power vs. (29. 76±2.53)/400 power, (47. 64±2.06)/400 power vs. (29.6±1. 61)/400 power and (51. 05±4. 23)/400 power vs.(33.44±2.06)/400 power, all P＜0. 05]. Conclusions The velcade could decrease the expressions of the NF-κBp65 and IL-1β and diminish the neuronal apoptosis. The neuroprotective mechanism of velcade may lie in decreasing apoptosis through inhibiting inflammation.

Objective To evaluate clinical value of double contrast-enhanced ultrasonography(DCUS)in diagnosing gastric stromal tumors(GST). Methods The medical records of 26 patients with a histological diagnosis of GST were retrospectively reviewed. The correlation between DCUS features and pathological findings of the lesions was compared. Results Total 26 cases of GST were divided into low risk group( 16 cases) and high risk group (10 cases). The size,contour,liquefactive necrosis and enhancement distribution of lesions were markedly correlated with the pathobiologic behaviour of the tumor ( P ＜0.05). There was no significant difference in border and metastasis of lesions between two groups( P ＞0.05). It was shown that DCUS supplied statistically significant improvement in the localization diagnosis and detection of liquefactive necrosis versus oral contrast-enhanced ultrasonography ( P ＜ 0.05 ). Conclusions DCUS is considered as a valuable tool in diagnosing location as well as pathobiological behaviour of GST. It can provide the guidance and reference comments for treatment algorithms.

There is growing interest in biodiesel and this results in the accumulation of glycerol. The exploitation and application of glycerol has attracted more and more attention. In the current study, glycerol was biotransformed to produce 3-hydroxypropionaldehyde by genetic engineering bacteria. It is known that 3-hydroxypopionaldehyde has been widely used as an important intermediate for chemicals, effective antimicrobial agent, and fix agent for tissues. A pair of primers was designed on the basis of the sequence of both NH2-terminus and the amino acid sequence of glycerol dehydratase reported by NCBI, and a fragment about 1.6 kb was obtained by PCR amplification using the total genome DNA of Lactobacillus reuteri as template, then the fragment was cloned to the pMD18-T vector and sequenced. Two specific primers were designed according to the obtained sequence, and a fragment with length of 1674 bp was amplified using PCR with these two specific primers. Consequently, the resulting products were digested with EcoR I and Hind III and ligated using T4 DNA ligase to the pET28b vector digested with the same enzymes. The recombinant plasmid, named pET28b-dhaB, was transformed into E. coli BL21. The positive clones were induced with IPTG and the expression products were further analyzed by SDS-PAGE, indicating that protein with a molecule weight of around 65 kD was obtained. Furthermore, the glycerol dehydratase activity was evaluated and compared with the wild type strain as well.
Cloning, Molecular ; Escherichia coli ; genetics ; metabolism ; Glyceraldehyde ; analogs & derivatives ; chemistry ; metabolism ; Hydro-Lyases ; biosynthesis ; genetics ; Lactobacillus reuteri ; enzymology ; genetics ; Propane ; chemistry ; metabolism ; Recombinant Proteins ; biosynthesis ; genetics

Objective To discuss the application of prospective risk nursing measures in reducing the occurrence of postoperative local hematoma at femoral artery puncture site. Methods A total of 605 patients receiving femoral artery puncture for interventional management were collected as the control group , and 322 patients were used as the intervention group. The risk factors causing postoperative local hematoma at femoral artery puncture site in the control group were recorded, and based on which risk estimation survey table was designed. According to risk estimation survey, the prospective intervention measures, including risk assessment, standardization of professional training, individual health education, the improvement of care appliance, etc. were carried out for the patients of the intervention group. The occurrences of postoperative local hematoma at femoral artery puncture site were recorded and the results were compared between the two groups. Results No statistically significant differences in demographic and clinical data existed between the two groups. However, the incidence of hematoma in the intervention group was significantly lower than that in the control group (χ2 = 4.652, P < 0.05), although the difference in the severity of hematoma was not significant between the two groups. Conclusion The use of prospective risk nursing measures can effectively reduce the incidence of postoperative hematoma at femoral artery puncture site.

usions DCUS could initial estimate the pathological typing of advanced gastric caner before operation.

Objective To investigate the effect of Sirt1 gene knockout on chronic kidney disease induced by 5/6 nephrectomy in mice and vascular endothelial growth factor (VEGF)/fetal liver kinase-1 (Flk-1) signaling pathway.Methods Twenty four male Sirt1 +/+ and Sirt1 +/-mice wererandomly divided into four groups:Sirt1+/+ mice with sham-operation (WT-Sham,n=6),Sirt1+/-mice with sham-operation (KO-Sham,n=6),Sirt1 +/+ mice with 5/6 nephrectomy (WT-Nx,n=6) and Sirt1 +/-mice with 5/6 nephrectomy (KO-Nx,n=6).Proteinuria was determined by urine collection from 8:00 to 8:00 the next day at 20 weeks.Serum creatinine (Scr),urea nitrogen (BUN) and the renal pathological changes were measured after 20 weeks.Expressions of Sirt1,collagen Ⅰ and transforming growth factor β(TGF-β) were used to analyze the changes of renal fibrosis by immunohistochemistry staining.Real-time PCR and Western blotting were used to measure the mRNA and protein expressions of Sirt1,fibronectin,collagen Ⅰ,VEGF and Flk-1 in kidney.Results Sirt1 expressed in glomernlar endothelial cells,podocytes,mesangial cells and renal tubular epithelial cells in Sirt1 +/+ mice,while Sirt1 expression intensity was significantly reduced in Sirt1 +/-mice.Compared with the WT-Sham group,WT-Nx group had increased proteinuria,BUN,Scr,glomernlar sclerosis index and tubulointerstitial fibrosis index at 12 weeks after operation (all P ＜ 0.01),and KO-Nx group had exacerbated the above up-regulations (all P ＜ 0.01).Compared with those in WT-Sham group,the expressions of fibronectin,collagen Ⅰ and TGF-β were up-regulated in WT-Nx group (all P ＜ 0.01),and were significantly augmented in KO-Nx group (all P ＜ 0.01).Compared with those in WT-Sham group,renal mRNA and protein expressions of VEGF and Flk-1 were decreased in WT-Nx group,and KO-Nx group aggravated their down-regulation (all P ＜ 0.01).Conclusions Sirt1 gene knockout can increase proteinuria and Scr,and aggravate renal pathology and renal fibrosis in 5/6 nephrectomized mice,which is associated with the inhibition of VEGF/Flk-1 signaling pathway.It is suggested that Sirt1 may be a potential therapeutic target of chronic kidney disease.

Nitriles are an important type of synthetic intermediates in the production of fine chemicals because of their easy preparations and versatile transformations. The traditional chemical conversion of nitriles to carboxylic acids and amides is feasible but it requires relatively harsh conditions of heat, acid or alkali. Nitrile converting enzymes (nitrilase, nitrile hydratase and amidase) which are used as biocatalyst for the production of fine chemicals have attracted substantial interest because of their ability to convert readily available nitriles into the corresponding higher value amides or acids under mild conditions with excellent chemo-, regio- and stereo-selectivities. Many nitrile converting enzymes have been explored and widely used for the production of fine chemicals. In this paper, various examples of biocatalytic synthesis of pharmaceuticals and their intermediates, agrochemicals and their intermediates, food and feed additives, and other fine chemicals are presented. In the near future, an increasing number of novel nitrile converting enzymes will be screened and their potential in the production of useful fine chemicals will be further exploited.
Amides ; metabolism ; Amidohydrolases ; metabolism ; Aminohydrolases ; metabolism ; Carboxylic Acids ; metabolism ; Chemical Industry ; methods ; Hydro-Lyases ; metabolism ; Nitriles ; chemistry

Objective:To evaluate the screening value of serum pepsinogen (PG) Ⅰ, pepsinogen ratio (PGR, PG Ⅰ/PG Ⅱ) and gastrin 17 (G17) levels combined with gastroscopy for early-stage gastric cancer in high incidence areas of gastric cancer in Qinghai Province.Methods:A total of 2 700 cases were identified as the appropriate age (40-69 years) target population through the questionnaire survey from 25 000 local residents in high incidence areas of gastric cancer in Qinghai Province. The serum PGⅠ, PGⅡ and G17 levels of the 2 700 target population were determined by ELISA, and PGR were calculated. And then 949 patients with abnormal levels of PG and G17 were screened out as a high-risk group of gastric cancer to receive gastroscopy and pathologic biopsy. According to the results of gastroscopy and biopsy, the patients were divided into non-atrophic gastritis group, atrophic gastritis group, peptic ulcer group, early-stage gastric cancer group, and advanced gastric cancer group. The optimal threshold and its sensitivity and specificity of serum PG Ⅰ, PGR and G17 levels for diagnosis of early-stage and advanced gastric cancer were determined based on the receiver operator characteristic curve (ROC).Results:Totally 949 cases received gastroscopy and 649 cases received pathological biopsy, including 239 cases of non-atrophic gastritis, 500 cases of atrophic gastritis, 197 cases of peptic ulcer, 5 cases of early-stage gastric cancer, and 8 cases of advanced gastric cancer. The level of serum PG Ⅰ in the early-stage gastric cancer group (70.00±12.35 μg/L) and advanced gastric cancer group (38.39±2.77 μg/L) was significant lower than that in the non-atrophic gastritis group (103.89±37.45 μg/L, both P<0.05), and the value of early-stage gastric cancer group was obviously higher than that of advanced gastric cancer group ( P<0.05). The PGR of the early-stage gastric cancer group (3.74±1.40) and the advanced gastric cancer group (2.05±0.59) was significantly lower than that in the non-atrophic gastritis group (9.18±4.10, both P<0.05), and the value of early-stage gastric cancer group was significantly higher than that of the advanced gastric cancer group ( P<0.05). The level of serum G17 in the early gastric cancer group (18.03±4.52 pmol/L) and the advanced gastric cancer group (25.15±3.76 pmol/L) was significantly higher than that in the non-atrophic gastritis group (14.99±7.12 pmol/L, both P<0.05), and the level of early-stage gastric cancer group was significantly lower than that of advanced gastric cancer group ( P<0.05). According to the analysis of ROC in the diagnosis of early-stage gastric cancer, the best threshold of PG Ⅰ, PGR and G17 was 71.85 μg/L, 5.04, and 15.65 pmol/L, respectively, and the corresponding sensitivity and specificity was 80.0% and 59.0%, 100.0% and 70.4%, and 80.0% and 69.3%, respectively, for PG Ⅰ, PGR and G17. The analysis of ROC in the diagnosis of advanced gastric cancer showd that the best critical value of PG Ⅰ, PGR and G17 was 42.55 μg/L, 2.79 and 20.55 pmol/L, respectively, and the corresponding sensitivity and specificity was 100.0% and 95.3%, 100.0% and 92.1%, and 100.0% and 89.7%, respectively. Conclusion:Using serological detection of PG and G17 to screen high-risk group of gastric cancer, and then making diagnosis by gastroscopy and biopsy is an effective, low-cost and non-invasive approach for the early-stage gastric cancer in high incidence areas of gastric cancer in Qinghai Province.

OBJECTIVE To identify the potential pathogenic mutation in a Chinese family with split hand/foot malformation (SHFM). METHODS Affymetrix SNP 6.0 array was used to perform a genome-wide copy number variations scan, and quantitative real-time PCR (qPCR) was applied to validate the identified genomic duplication. RESULTS A ~560 kb microduplication on the chromosome 10q24 was identified. The qPCR assay confirmed the presence of this microduplication in all the available affected family members. CONCLUSION The ~560 kb microduplication is probably the pathogenic mutation underlying the SHFM phenotype in the studied family.
Adolescent ; Adult ; Asian Continental Ancestry Group ; genetics ; China ; Chromosome Duplication ; Chromosomes, Human, Pair 10 ; genetics ; DNA Copy Number Variations ; Foot Deformities, Congenital ; genetics ; Hand Deformities, Congenital ; genetics ; Humans ; Male ; Middle Aged ; Pedigree ; Young Adult

Objective:To assess the clinical significance of next-generation sequencing (NGS)-based IGH/ IGK gene rearrangement analysis versus flow cytometry (FCM) in diagnosing minimal residual disease (MRD) of children with acute B-cell lymphoblastic leukemia (B-ALL). Methods:Clinical data, NGS-MRD and FCM-MRD findings at the initial diagnosis and after induction chemotherapy of 85 children diagnosed as B-ALL in Children′s Hospital of Nanjing Medical University from July 2019 to July 2021, were retrospectively analyzed.The sensitivity of the two methods, and the positive rate were compared by χ2 test or Fisher′ s test.The correlation was identified by Spearman correlation analysis. Results:Dominant clone sequences were detected in all children at the initial diagnosis by NGS, while selection markers were identified by FCM in 75(88.2%) patients.Positive MRD rate detected by NGS-MRD was significantly higher than that of FCM-MRD at the same time point after induction chemotherapy[31.8%(27/85) vs.9.4%(8/85), P<0.001]. Compared with those of FCM-MRD, NGS-MRD had good sensitivity (100.0%), specificity (75.3%) and negative predictive value (100.0%), and the positive predictive value was 29.6%.MRD results detected by NGS were consistent with that of FCM ( r=0.569, P<0.001). By July 27, 2022, 2 patients with NGS-MRD (+ )FCM-MRD (-)relapsed during maintenance chemotherapy. Conclusions:NGS is highly consistent with FCM in the detection of MRD in children with B-ALL, which is more sensitive.The combination of NGS-MRD and FCM-MRD benefits more in monitoring MRD in children with B-ALL after induction chemotherapy.