Engineering the plastic degradation enzyme Ple629 from marine consortium to improve its thermal stability / 生物工程学报

Petrochemical-derived polyester plastics such as polyethylene terephthalate (PET) and polybutylene adipate terephthalate (PBAT) have been widely used. However, the difficulty to be degraded in nature (PET) or the long biodegradation cycle (PBAT) resulted in serious environmental pollution. In this connection, treating these plastic wastes properly becomes one of the challenges of environment protection. From the perspective of circular economy, biologically depolymerizing the waste of polyester plastics and reusing the depolymerized products is one of the most promising directions. Recent years have seen many reports on polyester plastics degrading organisms and enzymes. Highly efficient degrading enzymes, especially those with better thermal stability, will be conducive to their application. The mesophilic plastic-degrading enzyme Ple629 from the marine microbial metagenome is capable of degrading PET and PBAT at room temperature, but it cannot tolerate high temperature, which hampers its potential application. On the basis of the three-dimensional structure of Ple629 obtained from our previous study, we identified some sites which might be important for its thermal stability by structural comparison and mutation energy analysis. We carried out transformation design, and performed expression, purification and thermal stability determination of the mutants. The melting temperature (Tm) values of mutants V80C and D226C/S281C were increased by 5.2 ℃ and 6.9 ℃, respectively, and the activity of mutant D226C/S281C was also increased by 1.5 times compared with that of the wild-type enzyme. These results provide useful information for future engineering and application of Ple629 in polyester plastic degradation.

Expression, purification and characterization of a novel bis (hydroxyethyl) terephthalate hydrolase from Hydrogenobacter thermophilus / 生物工程学报

PET (polyethylene terephthalate) is one of the most important petrochemicals that is widely used in mineral water bottles, food and beverage packaging and textile industry. Because of its stability under environmental conditions, the massive amount of PET wastes caused serious environmental pollution. The use of enzymes to depolymerize PET wastes and upcycling is one of the important directions for plastics pollution control, among which the key is the depolymerization efficiency of PET by PET hydrolase. BHET (bis(hydroxyethyl) terephthalate) is the main intermediate of PET hydrolysis, its accumulation can hinder the degradation efficiency of PET hydrolase significantly, and the synergistic use of PET hydrolase and BHET hydrolase can improve the PET hydrolysis efficiency. In this study, a dienolactone hydrolase from Hydrogenobacter thermophilus which can degrade BHET (HtBHETase) was identified. After heterologous expression in Escherichia coli and purification, the enzymatic properties of HtBHETase were studied. HtBHETase shows higher catalytic activity towards esters with short carbon chains such as p-nitrophenol acetate. The optimal pH and temperature of the reaction with BHET were 5.0 and 55 ℃, respectively. HtBHETase exhibited excellent thermostability, and retained over 80% residual activity after treatment at 80 ℃ for 1 hour. These results indicate that HtBHETase has potential in biological PET depolymerization, which may facilitate the enzymatic degradation of PET.

Effect of FTY-720 on Pulmonary Fibrosis in Mice via the TGF-β1Signaling Pathway and Autophagy

We investigated whether FTY-720 might have an effect on bleomycin-induced pulmonary fibrosis through inhibiting TGF-β1 pathway, and up-regulating autophagy. The pulmonary fibrosis was induced by bleomycin. FTY-720 (1 mg/kg) drug was intraperitoneally injected into mice. Histological changes and inflammatory factors were observed, and EMT and autophagy protein markers were studied by immunohistochemistry and immunofluorescence. The effects of bleomycin on MLE-12 cells were detected by MTT assay and flow cytometry, and the related molecular mechanisms were studied by Western Blot. FTY-720 considerably attenuated bleomycin-induced disorganization of alveolar tissue, extracellular collagen deposition, and α-SMA and E-cadherin levels in mice. The levels of IL-1β, TNF-α, and IL-6 cytokines were attenuated in bronchoalveolar lavage fluid, as well as protein content and leukocyte count. COL1A1 and MMP9 protein expressions in lung tissue were significantly reduced. Additionally, FTY-720 treatment effectively inhibited the expressions of key proteins in TGF-β1/TAK1/P38MAPK pathway and regulated autophagy proteins. Similar results were additionally found in cellular assays with mouse alveolar epithelial cells. Our study provides proof for a new mechanism for FTY-720 to suppress pulmonary fibrosis. FTY-720 is also a target for treating pulmonary fibrosis.

Clinical and endoscopic characteristics of adult celiac disease / 中华内科杂志

Objective:The study aimed to analyze the clinical and endoscopic characteristics of adult celiac disease (CD) to provide a scientific basis for more effective CD diagnosis and treatment.Methods:In this cross-sectional study, the clinical and endoscopic data of 96 adult CD patients treated in the Department of Gastroenterology of the People′s Hospital of Xinjiang Uygur Autonomous Region from March 2016 to December 2021 were retrospectively collected and analyzed.Results:A total of 96 CD patients were diagnosed, including 33 men and 63 women. The average age was 47±14 years (range, 18-81 years). The disease occurred mainly in the age group of 31-60 years. The median course of the disease was 2.0 (0.2-40.0) years. There were 41 (42.7%) classical and 55 (57.3%) non-classical CD patients. All patients with classical CD showed chronic diarrhea, often accompanied by abdominal pain (46.3%, 19/41), abdominal distension (17.1%, 7/41), anemia (65.9%, 27/41), and chronic fatigue (48.8%, 20/41). The main manifestations of non-classical CD were chronic abdominal pain (58.2%, 32/55), abdominal distension (32.7%, 18/55), anemia (40.0%, 22/55), and osteopenia/osteoporosis (38.2%, 21/55). Compared with non-classical CD, anemia developed more frequently in classical CD, and the difference was statistically significant ( P = 0.012). The incidence of complications in CD patients was 36.5% (35/96), and the main complications were thyroid disease (19.8%, 19/96), connective tissue disease (6.2%, 6/96), and kidney disease (6.2%, 6/96). There was no significant difference between classical and non-classical CD ( P>0.05). The frequency of endoscopic manifestations in CD patients was 84.4% (81/96). Duodenal bulb endoscopy showed nodular changes (72.9%, 70/96), grooved changes (10.4%, 10/96), and focal villous atrophy (9.4%, 9/96). The main manifestations of descending endoscopy were the decrease, flattening, or disappearance of duodenal folds (43.8%, 42/96), scallop-like changes (38.5%, 37/96), and nodular changes (34.4%, 33/96). Conclusions:Adult CD patients are mostly female. CD occurred mainly in the age group of 31-60 years. The clinical manifestations were mainly those of non-classical CD. Some patients often had other autoimmune diseases. Patients with characteristic endoscopic manifestations should be warned about the possibility of developing CD. Clinicians should strengthen the understanding of CD and reduce the related rates of missed diagnosis.

Function of AXL and molecular mechanisms in regulation of nasopharyngeal carcinoma / 中南大学学报(医学版)

OBJECTIVES@#Nasopharyngeal carcinoma (NPC) is a highly invasive epithelial malignant tumor with unique geographical and ethnic distribution characteristics. NPC is mostly found in south China and Southeast Asia, and its treatment mainly depends on radiotherapy and chemotherapy. However, NPC is usually found in the late stage, and local recurrence and distant metastasis are common, leading to poor prognosis. The receptor tyrosine kinase AXL is up-regulated in various tumors and it is involved in tumor proliferation, migration, invasion, and other processes, which are associated with poor prognosis of tumors. This study aims to detect the expression of AXL in NPC cell lines and tissues, and to investigate its biological function of AXL and the underlying molecular mechanisms in regulation of NPC.@*METHODS@#The expression levels of AXL in normal nasopharyngeal epithelial tissues and NPC tissues were analyzed by GSE68799, GSE12452, and GSE53819 data sets based on Gene Expression Omnibus (GEO) database. The Cancer Genome Atlas (TCGA) database was used to analyze the relationship between AXL and prognosis of head and neck squamous cell carcinoma (HNSC). The indicators of prognosis included overall survival (OS), disease-free interval (DFI), disease-specific survival (DSS), and progression-free interval (PFI). Western blotting assay was used to detect the AXL protein expression levels in normal nasopharyngeal epithelial cell line and NPC cell lines. Immunohistochemical method was used to detect AXL expression levels in normal nasopharyngeal epithelial tissues and NPC tissues. Cell lines with stable AXL knockdown were established by infecting 5-8F and Fadu cells with lentivirus interference vector, and cell lines with stable AXL overexpression were established by infecting C666-1 and HK-1 cells with lentivirus expression vector. Real-time PCR and Western blotting were used to detect the efficiency of knockdown and overexpression in stable cell lines. The effects of AXL knockdown or overexpression on proliferation, migration, and invasion of NPC cells were detected by CCK-8, plate colony formation, and Transwell assays, and the effect of AXL knockdown on tumor growth in nude mice was detected by subcutaneous tumor formation assay. The sequence of AXL upstream 2.0 kb promoter region was obtained by UCSC online database. The PROMO online database was used to predict AXL transcription factors with 0% fault tolerance, and the JASPAR online database was used to predict the binding sites of ETS1 to AXL. Real-time PCR and Western blotting were used to detect the effect of ETS1 on AXL protein and mRNA expression. The AXL upstream 2.0 kb promoter region was divided into 8 fragments, each of which was 250 bp in length. Primers were designed for 8 fragments. The binding of ETS1 to AXL promoter region was detected by chromatin immuno-precipitation (ChIP) assay to determine the direct regulatory relationship between ETS1 and AXL. Rescue assay was used to determine whether ETS1 affected the proliferation, migration, and invasion of NPC cells through AXL.@*RESULTS@#Bioinformatics analysis showed that AXL was highly expressed in NPC tissues (P<0.05), and AXL expression was positively correlated with OS, DFI, DSS, and PFI in HNSC patients. Western blotting and immunohistochemical results showed that AXL was highly expressed in NPC cell lines and tissues compared with the normal nasopharyngeal epithelial cell line and tissues. Real-time PCR and Western blotting results showed that knockdown and overexpression efficiency in the stable cell lines met the requirements of subsequent experiments. The results of CCK-8, plate colony formation, Transwell assays and subcutaneous tumor formation in nude mice showed that down-regulation of AXL significantly inhibited the proliferation, migration, invasion of NPC cells and tumor growth (all P<0.05), and the up-regulation of AXL significantly promoted the proliferation, migration, and invasion of NPC cells (all P<0.05).As predicted by PROMO and JASPAR online databases, ETS1 was a transcription factor of AXL and had multiple binding sites in the AXL promoter region. Real-time PCR and Western blotting results showed that knockdown or overexpression of ETS1 down-regulated or up-regulated AXL protein and mRNA expression levels. ChIP assay result showed that ETS1 bound to AXL promoter region and directly regulate AXL expression. Rescue assay showed that AXL rescued the effects of ETS1 on proliferation, migration and invasion of NPC cells (P<0.05).@*CONCLUSIONS@#AXL is highly expressed in NPC cell lines and tissues, which can promote the malignant progression of NPC, and its expression is regulated by transcription factor ETS1.

Study on vitamin D levels in serum and intestinal mucosa of 37 adult patients with celiac disease / 中华消化杂志

Objective:To analyze the expression of 25-hydroxyvitamin D [25(OH)D] in serum and vitamin D receptor (VDR) in descending duodenum of adult patients with celiac disease and the correlation between 25 (OH) D, VDR and the clinical characteristics and indexes of celiac disease.Methods:From September 1, 2020 to May 1, 2022, 37 patients who were diagnosed with celiac disease (celiac disease group) in the Department of Gastroenterology of People′s Hospital of Xinjiang Uygur Autonomous Region were enrolled. During the same period, according to gender, age and nationality matched at a ratio of 1 to 1, 37 patients who visited the hospital with suspected symptoms of celiac disease and finally were excluded from the diagnosis of celiac disease after screening (non-celiac disease group) were selected. General data and clinical characteristics of all the patients were collected, including diarrhea, bone mineral density (BMD), Marsh stage. Serum 25(OH)D levels were detected, and the expression of VDR(high or low expression) in the descending duodenum was evaluated by immunohistochemical staining. Independent sample t test, chi-square test and Spearman correlation analysis were used for statistical analysis. Results:The serum 25(OH)D level and the proportion of patients with high VDR expression in the celiac disease group were both lower than those of the non-celiac disease group ((12.40±8.93) μg/L vs. (16.78±7.09) μg/L; 45.9%, 17/37 vs. 75.7%, 28/37), and the proportion of patients with diarrhea was higher than that of the non-celiac disease group (45.9%, 17/37 vs. 21.6%, 8/37), and the differences were statistically significant ( t=-2.52, χ2=6.86 and 4.89, all P<0.05). The serum 25(OH)D level was positively correlated with the VDR expression level in the descending duodenum both in the celiac disease group and the non-celiac disease group ( r=0.75 and 0.64, both P<0.001), and was not correlated with the diarrhea symptoms in the 2 groups (both P>0.05). There was a positive correlation between serum 25(OH)D and BMD in the celiac disease group ( r=0.48, P=0.017), and serum 25(OH)D was also correlated with BMD in the non-celiac disease group ( r=0.93, P<0.001). The VDR expression level in the descending duodenum was negatively correlated with the Marsh stage in the celiac disease group ( r=-0.36, P=0.031), while the VDR expression level was not related to the Marsh stage in the non-celiac disease group ( P>0.05). Conclusions:Vitamin D metabolism imbalance exists in the serum and duodenal mucosa of adult patients with celiac disease, which is related to the severity of osteoporosis and histopathology. It is suggested that patients with celiac disease should receive vitamin D metabolism regulation treatment.

Chemotherapy and stroke in cancer patients / 国际脑血管病杂志

There is a certain relationship between chemotherapy and stroke in cancer patients. Its mechanism may be associated with the increase of the prevalence of traditional vascular factors, the promotion of coagulation dysfunction, the induction of anemia, the impairment of cardiac function, and vascular inflammation. The pathophysiological mechanism of chemotherapy-associated stroke is still in the exploratory stage. This article reviews the pathophysiological mechanism, monitoring indicators, and diagnosis and treatment progress of stroke in cancer patients during chemotherapy.

Structure-based engineering of PET hydrolase from Ideonella sakaiensis / 生物工程学报

Polyethylene terephthalate (PET) is a synthetic polymer consisting of ester bond-linked terephthalate and ethylene glycol. Tremendous amounts of PET have been produced and majority of them enters terrestrial and marine environment as wastes, posing serious threats to the global ecosystems. In 2016, a PET hydrolase from a PET-assimilating bacterium Ideonalla sakaiensis was reported and termed as IsPETase. This enzyme outperforms other PET-hydrolyzing enzymes in terms of its PET hydrolytic activity at ambient temperature, thus holds a great promise for PET biodegradation. In order to improve IsPETase activity, we conducted structure-based engineering to modify the putative substrate-binding tunnel. Among the several variants to the N233 residue of IsPETase, we discovered that the substitution of N233 with alanine increases its PET hydrolytic activity, which can be further enhanced when combined with a R280A mutation. We also determined the X-ray crystal structure of the IsPETase N233A variant, which shares nearly identical fold to the WT protein, except for an open end of subsite Ⅱ. We hypothesized that the smaller side chain of N233A variant might lead to an extended subsite Ⅱ for PET binding, which subsequently increases the enzymatic activity. Thus, this study provides new clues for further structure-based engineering of PETase.

Clinical efficacy of platelet rich plasma combined with Bio-oss artificial bone powder in repairing bone defects around implant / 中国基层医药

Objective@#To research the clinical effect of platelet rich plasma combined with Bio-oss artificial bone meal in repairing bone defects around implants.@*Methods@#From January 2015 to January 2017, 70 cases who completed the implant repair in Lishui People's Hospital, and later confirmed the defects around the implant were selected in the research.They were equally divided into observation group and control group according to admission, with 35 cases in each group.The control group was treated with Bio-oss artificial bone powder.The observation group was treated with platelet rich plasma combined with Bio-oss artificial bone powder.The clinical effects of the two groups were compared and analyzed.@*Results@#Before treatment, the probe depth (PD), clinical attachment loss (CAL), plaque index (PI), gingival sulcus hemorrhage index (SBI) of the observation group and the control group were (7.18±1.31)mm, (5.68±0.87)mm, (1.23±0.31), (2.48±0.34) and (7.22±1.28)mm, (5.74±0.83)mm, (1.27±0.33), (2.53±0.36), respectively, which were significantly higher than those after treatment (all P<0.05). Before treatment, the bone mineral density (BMD) levels of the two groups were (35.48±4.58)g/cm3 and (35.74±4.61)g/cm3, respectively, which were significantly lower than those after treatment (all P<0.05). The levels of PD, CAL, PI and SBI in the observation group at 3 months and 6 months after treatment were (4.48±1.01)mm, (4.27±0.53)mm, (0.68±0.21), (1.12±0.13) and (3.43±0.76)mm, (3.58±0.54)mm, (0.38±0.07), (0.41±0.08), respectively, which were significantly lower than those in the control group [(5.19±0.83)mm, (4.83±0.71)mm, (0.89±0.27), (1.46±0.21) and (4.14±0.98)mm, (4.21±0.62)mm, (0.49±0.11), (0.63±0.13)], the differences were statistically significant (t=3.213, 3.739, 3.632, 8.144 and 3.387, 4.533, 4.991, 8.526, P=0.002, 0.000, 0.000, 0.000, 0.001, 0.000, 0.000, 0.000, 0.000, 0.000). The levels of TA anterior temporalis muscle EMG and MA masseter muscle EMG in the two groups at 6 months after treatment were (76.81±15.17)mV, (67.56±14.43)mV, (74.24±13.28)mV, (68.27±13.03)mV, (66.44±13.28)mV, (58.76±13.07)mV, (65.58±11.55)mV, (59.04±10.76)mV, respectively, which were significantly higher than those before treatment [(46.48±11.15) mV, (38.19±13.04) mV, (54.83±12.57) mV, (56.48±13.13) mV and (45.71±10.98) mV, (38.23±13.11)mV, (55.14±12.69) mV, (56.61±13.08) mV] (all P<0.05). After treatment, the ICP-HB, ICMAP-MC, TA, EMG levels in the observation group were significantly higher than those in the control group, the differences were statistically significant (t=3.042, 2.674, 2.911, 2.994, P=0.003, 0.009, 0.004 and 0.003).@*Conclusion@#The effect of rich plate plasma combined with Bio-oss artificial bone powder on the repair of bone defect around the implant is significant.The platelet rich plasma can speed up and improve the repair speed and repair quality of bone defect around the implant, so that the therapeutic effect of implant repair can be maintained for a long time.

Interpretation of clinical practice guideline for anorectal day surgery 2019 edition / 中华胃肠外科杂志

As the rapid development of minimally invasive techniques, anesthesia, and enhanced recovery after surgery (ERAS), anorectal day surgery receiving more and more attention by improving efficiency of medical care while reducing cost and hospitalized infection. However, day surgery also faces the challenge of completing the whole process from patient admission to discharge within 24 hours. Therefore, establishing a reasonable and detailed day surgery process is the cornerstone to guarantee safe medical practice and patients satisfaction. National Clinical Research Center for Geriatric Disorders (Xiangya), together with China Ambulatory Surgery Alliance formulates the clinical practice guideline for anorectal day surgery 2019 edition. Here we make some interpretations of the guidelines on the detailed process of anorectal day surgery, including indication, preoperative examination, preoperative risk evaluation, health education, assessment of day surgery anesthesia and before leaving postanesthesia care unit (PACU), postoperative management, assessment of discharge and follow-up, for the convenience of various medical centers.

Association between circular RNAs expression in serum and gastric cancer / 中华流行病学杂志

Objective@#To analyze the association between circular RNAs expression in serum and gastric cancer and evaluate the potential of the related markers in early diagnosis of gastric cancer.@*Methods@#Forty eight gastric cancer cases in Linqu County People’s Hospital were selected as case group, and 48 controls matched by age and sex were randomly selected in the gastric cancer screening cohort during the same period. The expression levels of hsa_circ_002059, hsa_circ_0000096 and hsa_circ_0001895 were detected by quantitative real-time PCR. The results were compared between case group and control group.@*Results@#The positive expression rates of hsa_circ_002059, hsa_circ_0000096 and hsa_circ_0001895 were 70.8%, 47.9%, 75.0% in case group, slightly higher than those in control group (58.3%, 31.3%, 60.4%), although P values were all more than 0.05. The expression level medians of the 3 candidate circRNAs expression levels were 1.60% (0-5.64%), 0 (0-0.61%), 0.91% (0.06%-1.88%) in case group, while 0.05% (0-6.07%), 0 (0-0.34%), 0.42% (0-1.39%) in control group, respectively. Conditional logistic regression analysis showed that the association strength of high expressions of 3 candidate circRNAs with gastric cancer showed an increase trend, but the differences had no significance after adjusted by Helicobacter pylori infection, smoking and drinking status (all P>0.05). Further analysis by combining the 3 candidate circRNAs showed the increased strength of association between circRNAs and gastric cancer with the elevated number of positive circRNAs in serum (trend test P=0.040) compared with circRNAs negative persons.@*Conclusion@#Our study preliminarily suggested that the expression of hsa_circ_002059, hsa_circ_0000096 and hsa_circ_0001895 in serum might be correlated with gastric cancer.

Clinical efficacy of platelet rich plasma combined with Bio-oss artificial bone powder in repairing bone defects around implant / 中国基层医药

Objective To research the clinical effect of platelet rich plasma combined with Bio-oss artificial bone meal in repairing bone defects around implants.Methods From January 2015 to January 2017,70 cases who completed the implant repair in Lishui People's Hospital,and later confirmed the defects around the implant were selected in the research.They were equally divided into observation group and control group according to admission,with 35 cases in each group.The control group was treated with Bio-oss artificial bone powder.The observation group was treated with platelet rich plasma combined with Bio-oss artificial bone powder.The clinical effects of the two groups were compared and analyzed.Results Before treatment,the probe depth (PD),clinical attachment loss (CAL),plaque index (PI),gingival sulcus hemorrhage index (SBI) of the observation group and the control group were (7.18 ± 1.31)mm,(5.68 ±0.87)mm,(1.23 ±0.31),(2.48 ±0.34) and (7.22 ± 1.28)mm,(5.74 ±0.83) mm,(1.27 ± 0.33),(2.53 ± 0.36),respectively,which were significantly higher than those after treatment (all P ＜ 0.05).Before treatment,the bone mineral density (BMD) levels of the two groups were (35.48 ± 4.58) g/cm3 and (35.74 ±4.61) g/cm3,respectively,which were significantly lower than those after treatment (all P ＜ 0.05).The levels of PD,CAL,PI and SBI in the observation group at 3 months and 6 months after treatment were (4.48 ± 1.01) mm,(4.27 ±0.53)mm,(0.68 ±0.21),(1.12 ±0.13) and (3.43 ±0.76) mm,(3.58 ±0.54)mm,(0.38 ±0.07),(0.41 ±0.08),respectively,which were significantly lower than those in the control group [(5.19 ± 0.83) mm,(4.83 ±0.71)mm,(0.89 ±0.27),(1.46 ±0.21) and (4.14 ±0.98)mm,(4.21 ±0.62)mm,(0.49 ±0.11),(0.63 ±0.13)],the differences were statistically significant (t =3.213,3.739,3.632,8.144 and 3.387,4.533,4.991,8.526,P =0.002,0.000,0.000,0.000,0.001,0.000,0.000,0.000,0.000,0.000).The levels of TA anterior temporalis muscle EMG and MA masseter muscle EMG in the two groups at 6 months after treatment were (76.81 ± 15.17) mV,(67.56 ±14.43)mV,(74.24 ± 13.28) mV,(68.27 ± 13.03) mV,(66.44 ± 13.28) mV,(58.76 ± 13.07) mV,(65.58 ±11.55) mV,(59.04 ± 10.76) mV,respectively,which were significantly higher than those before treatment [(46.48 ±11.15) mV,(38.19± 13.04) mV,(54.83 ±12.57) mV,(56.48 ±13.13) mV and (45.71 ±10.98) mV,(38.23 ±13.11)mV,(55.14±12.69) mV,(56.61 ± 13.08) mV] (all P ＜0.05).After treatment,the ICP-HB,ICMAP-MC,TA,EMG levels in the observation group were significantly higher than those in the control group,the differences were statistically significant (t =3.042,2.674,2.911,2.994,P =0.003,0.009,0.004 and 0.003).Conclusion The effect of rich plate plasma combined with Bio-oss artificial bone powder on the repair of bone defect around the implant is significant.The platelet rich plasma can speed up and improve the repair speed and repair quality of bone defect around the implant,so that the therapeutic effect of implant repair can be maintained for a long time.

A study on the association between vascular endothelial growth factor A polymorphisms and necrotizing enterocolitis / 中国新生儿科杂志

Objective To study the relationship between single nucleotide polymorphisms (SNPs) of vascular endothelial growth factor A (VEGFA) gene and neonatal necrotizing enterocolitis (NEC).Method From August 2014 to December 2016,preterm infants with a ≥ Ⅱ stage (Modified Bell staging criteria) of NEC admitted to our hospitals were assigned as NEC group.Preterm infants without NEC with similar gestational age and body weight during the same period were assigned as the control group.SNPs of VEGFA including rs1005230,rs833067,rs3025010,rs3025035,rs3025036,rs10434,and rs6905288 were analyzed using SEQUENOM MassARRAY platform and multiplex allele-specific PCR.The concentration of VEGFA in the plasma of the two groups was examined using enzyme-linked immunosorbent assay (ELISA).Result A total of 110 infants were reviewed,including 30 infants in NEC group and 80 infants in the control group.The results showed a significant association of the minor allele frequency (MAF) for T in rs1005230 and C in rs833067 with NEC.The frequencies of C/T (OR=4.810,95％CI 1.742～13.278) and C/T-T/T (OR=4.892,95％CI 1.801～13.246) genotypes in rs1005230,and frequencies of T/C (OR=4.373,95％CI 1.578～12.129) and T/C-C/C (OR=4.000,95％CI 1.484～10.828) genotypes in rs833067 were significantly higher in NEC group than the control group (P＜0.05).Infants with MAF in rs1005230 and rs833067 had significantly lower plasma level of VEGFA than infants without MAF (P＜0.01).Conclusion The SNPs of rs1005230 and rs833067 may be associated with lower level of VEGFA in plasma and higher risk for NEC.

Structure-function relationships of industrial enzymes / 生物工程学报

Industrial enzymes are the "chip" of modern bio-industries, supporting tens- and hundreds-fold of downstream industries development. Elucidating the relationships between enzyme structures and functions is fundamental for industrial applications. Recently, with the advanced developments of protein crystallization and computational simulation technologies, the structure-function relationships have been extensively studied, making the rational design and de novo design become possible. This paper reviews the progress of structure-function relationships of industrial enzymes and applications, and address future developments.

Effects of n-hexane on the serum level of three neurotoxicity biomarkers in rat / 中国职业医学

OBJECTIVE: To observe the effect of n-hexane subchronic exposure on the serum level of neuron specific enolase(NSE), neurofilament light chain protein(NF-L) and nerve growth factor(NGF) in rat, and to explore the feasibility of using NSE, NF-L and NGF as biomarkers of n-hexane neurotoxicity. METHODS: Specific pathogen free male SD rats were randomly divided into control group and low-, medium-and high-dose exposure groups, with 6 rats in each group. Rats in the low-, medium-and high-dose exposure groups were given n-hexane solution at doses of 168, 675 and 2 700 mg/kg body mass, respectively, while rats in the control group were gavaged with 0.9% sodium chloride solution, once a day, 5 days a week for 6 weeks. At week 0, 2, 4, and 6 of exposure, the body mass of the rats was weighed, the gait scores were performed, and the serum levels of NSE, NF-L, and NGF were detected.RESULTS: Body mass, gait score and serum levels of NSE and NF-L in rats were statistically significant in terms of the n-hexane exposure dose and exposure time(P<0.01). At the 6 th week of n-hexane exposure, the body mass of the three dose exposure groups was lower than that of the control group(P<0.05), and the gait score was higher than that of the control group(P<0.05). Moreover, the abnormal gait of the rats showed a dose-effect relationship with the increasing n-hexane poisoning dose. At week 2, 4 and 6, the serum levels of NSE and NF-L in these three dose exposure groups were higher than that in the control group(P<0.05). In addition, the serum level of NF-L in rats in the medium-and high-dose exposure groups increased with the n-hexane exposure time increasing and showed a time-effect relationship(P<0.05). The level of serum NGF in rats was statistically significant in the main effects of n-hexane dose and duration of exposure(P<0.05). The serum NGF level in the high-dose exposure group was lower than that in the control group, the low-dose and medium-dose exposure groups(P<0.05). NGF level in serum of rats at week 6 was lower than that at week 0, 2 and 4(P<0.05). CONCLUSION: Both NSE and NF-L in serum can be used as biomarkers for the early effect of n-hexane on peripheral nerve injury. The feasibility of using serum NGF as a biomarker for the early effect of n-hexane on peripheral nerve injury warrants further investigation.

Relationship between different types of Helicobacter pylori infection and progress of atrophic gastritis / 中华消化杂志

Objective To observe the distribution of different types of Helicobacter pylori (H.pylori)infection in non-atrophic gastritis,atrophic gastritis and intestinal metaplasia.Methods From January 2015 to January 2017,457 hospitalized patients with gastritis were collected.All the inpatients received 14C-urea breath test(14C-UBT), H.pylori antibody typing detection, anesthesia gastroendoscopy and pathological analysis of biopsies.According to the results of H.pylori antibody typing,all patients were divided into typeⅠ H.pylori infection group,typeⅡ H.pylori infection group and H.pylori negative group.The relation among different types of H.pylori infection,age and pathological type were analyzed.Analysis of variance and chi-square test were performed for statistical analysis.Results The mean age of patients in typeⅠH.pylori infection group(n=135),typeⅡ H.pylori infection group(n=98)and H.pylori negative group(n=224)in patients with non-atrophic gastritis was(47.5 ± 9.0),(49.0 ± 9.0)and(52.0 ± 11.0)years,respectively, which were lower than those of patients with atrophy gastritis((56.8 ± 10.3),(57.5 ± 12.4)and(62.6 ± 10.4) years,respectively)and patients with intestinal metaplasia((59.2 ± 11.1),(57.5 ± 12.6)and(57.8 ± 10.0)years,respectively),and the differences were statistically significant(F=17.90,6.82 and 18.30,all P<0.01).The proportion of patients with non-atrophic gastritis in H.pylori negative group,typeⅡ H.pylori infection group and typeⅠ H.pylori infection group decreased in turn(56.3%,126/224;40.8%,40/98 and 34.8%,47/135, respectively),and the differences were statistically significant(χ2 =18.44,P<0.01).The proportion of patients with intestinal metaplasia in H.pylori negative group,type Ⅱ H.pylori infection group and type Ⅰ H.pylori infection group increased in turn(23.2%,52/224;26.5%,26/98 and 42.2%,57/135),and the difference was statistically significant(χ2 =11.44,P=0.003).There was no significant difference in the proportion of patients with atrophic gastritis among H.pylori negative group,type Ⅱ H.pylori infection group and type Ⅰ H.pylori infection group(20.5%,46/224;32.7%,32/98 and 23.0%,31/135;χ2 = 5.60,P=0.061).Conclusions Gastric atrophy is easily induced in patients with H.pylori infection.In patients without symptoms,H.pylori should be eradicated before gastric atrophy,which can provide the greatest benefit to the patients.Different types of H.pylori infection have different role in the progress of atrophic gastritis.TypeⅠ H.pylori infection accelerate the progress of atrophic gastritis,especially from atrophic gastritis to intestinal metaplasia,which should be actively eradicated.

Efficacy and safety of simulated artificial pancreas in modulating stress hyperglycemia in critically ill patients:a prospective randomized controlled study / 中华危重病急救医学

Objective To explore efficacy and safety of simulated artificial pancreas in modulating stress hyperglycemia in critically ill patients. Methods A prospective randomized controlled study was performed. Seventy-two critically ill patients with stress hyperglycemia, aged 18-85 years, acute physiology and chronic health evaluationⅡ(APACHEⅡ) score over 15, two consecutive random blood glucose 11.1 mmol/L or higher, glycated hemoglobin (HbA1C) below 0.065, unable to eat food for 3 days after inclusion, or only accepting parenteral nutrition, admitted to intensive care unit (ICU) in Shanghai Punan Hospital of Pudong New District from January 1st, 2015 to June 30th, 2017 were enrolled. The patients were divided into three groups according to the random number table method, high-intensity group and low-intensity group were injected Novolin R (high-intensity group 2/3 dosage, low-intensity group 1/3 dosage) to modulate stress hyperglycemia by simulated artificial pancreas. Simulated artificial pancreas consisted of Guardian real time glucose monitoring system (GRT system), close-circle control algorithm and micro-pump;subcutaneous injection of Humulin 70/30 was applied to modulate stress hyperglycemia in humulin group. Real-time glucose levels of interstitial fluid in abdominal wall, equivalent to blood glucose levels, 10 minutes each time, were monitored by using of GRT system for all patients in three groups. Fasting serum levels of stress hormones including epinephrine and cortisol and insulin resistance index (IRI) were recorded within 24 hours after inclusion. Mean blood glucose, blood glucose variation coefficient, blood glucose target-reaching rate, blood glucose target-reaching time, hypoglycemia rate and 6-month mortality were measured. Twenty healthy adults from health administration department of the hospital were recruited as healthy control group. Results A total of 60 eligible critically ill patients were included in this study, each group with 20 patients. There was no significant difference in gender, age, APACHE Ⅱ scores among three groups. The levels of serum epinephrine, cortisol and IRI within 24 hours after inclusion in the three groups were significantly higher than those in healthy control group. The mean blood glucose levels of humulin group, low-intensity group, high-intensity group were decreased (mmol/L: 10.2±3.2, 8.4±2.6, 8.1±2.2), the blood glucose target-reaching rate were increased [40.2% (3 295/8 196), 71.1% (5 393/7 585), 80.4% (6 286/7 818)], the blood glucose target-reaching time were shortened (hours: 49.1±5.8, 24.6±4.6, 17.5±4.2), the hypoglycemia rates were increased respectively [1.3% (108/8 196), 2.8% (211/7 585), 4.0% (313/7 818)], with statistically significant differences (all 1 = 0.000). There was no significant difference in blood glucose variation coefficient and 6-month mortality among three groups [blood glucose variation coefficient: (29.4±3.7)%, (28.5±5.3)%, (26.1±4.6)%, 6-month mortality: 55.0%, 45.0%, 40.0%, all 1 > 0.05]. Conclusions Simulated artificial pancreas could effectively and safely modulate stress hyperglycemia in critically ill patients, high-intensity modulation could bring about better efficacy in the regulation of hyperglycemia. High-frequency blood glucose monitoring by using GRT system could promptly identify hypoglycemia and help it to be corrected.

Multifunctional Cervical Health Care Pillow / 中国医疗器械杂志

Cervical spondylosis is due to degenerative cervical disc and its stimulation or oppression of the adjacent nerves, spinal cord, spinal artery and other tissue caused by clinical symptoms. The cervical spine is an anatomical structure with activity, while the pillow has a certain plastic fixation effect on the cervical spine anatomy. Therefore, the pillow not only plays a health role in the cervical spine, but also plays an important role in restoring the normal physiological curvature of the cervical spine. Based on this, a multi-functional cervical vertebra treatment pillow is designed, which not only has the functions of traction, maintaining different positions of the cervical spine, correcting the cervical curvature and equipment exercises, but also has the functions of voice broadcast and network data terminal.

Application of iTRAQ multiplex tandem mass spectrometry in the expression of HGF in breast cancer cell invasion / 国际检验医学杂志

Objective To investigate the clinical value of iTRAQ multiplex tandem mass spectrometry in the detection of the ex-pression of hepatocyte growth factor (HGF) in patients with invasive breast cancer .Methods A total of 35 patients with breast cancer and 30 healthy subjects were selected from January 2014 to October 2016 in this hospital ,the expression of serum HGF in breast cancer patients with different clinical stages and healthy subjects was analyzed by iTRAQ labeling ,mass spectrometry ,library searching and Scqffold software ,and the differential expression of HGF was verified by Western blot .Results A total of 237 pro-teins were identified in the serum samples of this study ,and 89 proteins with strict quantitative criteria ,17 differentially expressed proteins ,included HGF ,were screened for breast cancer patients and healthy controls .iTRAQ markers showed that the expression level of serum HGF in different clinical stage of breast cancer patients was significantly higher than that in healthy subjects (P<0 .05) .The results of Western blot showed that the relative expression level of serum HGF in breast cancer patients was significant-ly higher than that in healthy subjects(P<0 .05) .Conclusion iTRAQ multiplex tandem mass spectrometry is useful for the detection of breast cancer patients with high expression of HGF ,which is of great significance in guiding the clinical treatment of breast cancer .

Establishment of a direct detection method for human platelet antigen1-17,Cab allele whole blood / 国际检验医学杂志

Objective To establish an acurate and convenient method to distinguish human platelet antigen(HPA) SNPs based on Target Enriched Multiplex-PCR(TEM-PCR),fluorescent probe melting curve analysis and blood direct PCR.Methods Design TEM-PCR primers and probes of HPA1-17,Cab alleles,amplify target sequences of all 18 alleles by blood direct PCR and distinguish different SNPs by melting curve of probes.Results The TEM-PCR could amplify all target sequences of 18 alleles and the melting curve analysis could distinguish those SNPs,the accuracy was equal to PCR-SSP method and the process was more convenient without blood genomic DNA extraction and subsequent gel electrophoresis thus decrease the cross-contamination risk.Conclusion Successfully established a HPA1-17,Cab alleles distinguishing method based on TEM-PCR,blood direct PCR and fluorescent probe melting curve analysis technique.