Immobilization of recombinant Escherichia coli cells expressing glucose isomerase using modified diatomite as a carrier for effective production of high fructose corn syrup in packed bed reactor.

To improve the operational stability of glucose isomerase in E. coli TEGI-W139F/V186T, the immobilized cells were prepared with modified diatomite as a carrier and 74.1% activity of free cells was recovered after immobilization. Results showed that the immobilized cells still retained 86.2% of the initial transformational activity after intermittent reused 40 cycles and the yield of D-fructose reached above 42% yield at 60 °C. Moreover, the immobilized cells were employed in the continuous production of High Fructose Corn Syrup (HFCS) in a recirculating packed bed reactor for 603 h at a constant flow rate. It showed that the immobilized cells exhibited good operational stability and the yield of D-fructose retained above 42% within 603 h. The space-time yield of high fructose corn syrup reached 3.84 kg L-1 day-1. The investigation provided an efficient immobilization method for recombinant cells expressing glucose isomerase with higher stability, and the immobilized cells are a promising biocatalyst for HFCS production.

Effect of Acupoint Catgut Embedding for Middle-Aged Obesity: A Multicentre, Randomised, Sham-Controlled Trial.

Objectives: This study aimed to examine the efficacy and safety of acupoint catgut embedding (ACE) for obesity over a 16-week treatment period using sham stimulation as the control. Methods: A multicenter, randomised, parallel, sham-controlled trial was conducted from February 10, 2017, to May 15, 2018. Men with waistlines ≥85 cm and women with ≥80 cm at three sites were randomised to receive eight sessions (over 16 weeks) of ACE (n = 108) or sham ACE (n = 108) with skin penetration at sham acupoints. The catgut was embedded once every two weeks using two alternating sets of acupoints. The follow-up lasted for an additional 24 weeks. The primary outcome was the percentage waistline reduction from baseline to week 16. Results: We included 216 individuals in the intention-to-treat analysis. At 16 weeks, the rate of waistline reduction was 8.80% (95% confidence interval (CI), 7.93% to 9.66%) in the ACE group and 4.09% (95% CI, 3.18% to 5.00%) in the sham control group, with a between-group difference of 4.71% (95% CI, 3.47% to 5.95%; P < 0.0001). This difference persisted throughout the entire follow-up period (between-group difference after 24-week additional weeks, 4.94% (95% CI, 3.58% to 6.30%); P < 0.001). The subgroup analyses of waistline by sex (male/female) revealed treatment effects of 1.93 (95% CI, -0.37 to 4.23, P = 0.1) in the male group and 3.19 (95% CI, 1.99 to 4.39, P < 0.001) in the female group. The adverse event analysis suggested that ACE and laboratory tests confirmed the safety of ACE. Discussion. ACE for 16 weeks could decrease the waistline and weight and was safe for the treatment of obesity. Further research is needed to evaluate the long-term efficacy and sex differences. This trial is registered with NCT02936973.

Enhanced catalytic efficiency and thermostability of glucose isomerase from Thermoanaerobacter ethanolicus via site-directed mutagenesis.

Glucose isomerase (GI) is a key enzyme in the preparation of high fructose corn syrup (HFCS). In this study, a mutant TEGI-M-L38 M/V137 L (TEGI-M2) of glucose isomerase (TEGI-M) originated from Thermoanaerobacter ethanalicus CCSD1 was obtained by site-directed mutagenesis. The TEGI-M2 showed an optimal activity at 85 ℃ and pH 6.5 with the divalent cations Co2+ and Mg2+. The structural differences between TEGI-M and TEGI-M2 were investigated based on the homology modeling and molecular docking, to elucidate the mechanism of improvement in the enzymatic properties. Compared with the original enzyme, the TEGI-M2 showed a 2.0-fold increased enzyme activity and a decreased Km from 234.2 mM to 85.9 mM. Finally, the application of mutant TEGI-M2 in HFCS one-step biosynthesis was attempted, resulting in a d-fructose yield of 67.3 %, which was 14.3 % higher than that of TEGI-M. This improved catalytic performance of TEGI-M2 was of great importance for the industrial preparation of d-fructose in one-step process.

Mining the Prognostic Value of HNRNPAB and Its Function in Breast Carcinoma.

Heterogeneous nuclear ribonucleoproteins (HNRNPs) are crucial members in the pathogenesis and progression of numerous cancers. However, the expression pattern and clinical significance of HNRNPs in breast carcinoma (BC) remain to be investigated. In the present study, bioinformatic analysis identified HNRNPAB as the only commonly upregulated HNRNP in BC. Elevated expression of HNRNPAB was positively associated with more aggressive diseases and poorer survival rates in BC. Pathway analysis revealed that HNRNPAB coexpressed genes were enriched in the pathway of G2/M phase transition, and the expression level of HNRNPAB was strongly correlated with those of CCNB1, CDK1, CDC25A, and CDC25C. Experiments in vitro demonstrated that HNRNPAB knockdown suppressed cell proliferation and blocked the G2/M phase transition in BC. Taken together, this study provides the initial evidence that HNRNPAB may be employed as an innovative therapeutic target as well as a prognostic biomarker in BC patients.

Clinical effect of catgut implantation at acupoints for the treatment of simple obesity: A multicentre randomized controlled trial.

BACKGROUND: Catgut implantation at acupoints (CIA) is a subtype of acupuncture that has been widely used to treat simple obesity, but evidence for its effectiveness remains scarce. The aim of this study is to evaluate the efficacy and safety of treating simple obesity with CIA. OBJECTIVE: This clinical trial aims to evaluate the effectiveness and safety of CIA used for treatment of simple obesity. METHODS: This is a multicentre, randomized, parallel, sham-controlled clinical trial. A total of 216 patients with simple obesity will be recruited. They will be randomly assigned in a 1:1 ratio to either the CIA group or the sham control group. All treatments will be given once every 2 weeks. The primary outcome measure is the rate of waistline reduction. Secondary outcome measures are the rates of reduction of body measurements, including weight, body mass index (BMI), hipline, waist-hip-ratio (WHR) and body fat percentage (BFP), the changes in scores on scales, including the Impact of Weight on Quality of Life Questionnaire (IWQOL-Lite), Short Form 36 (SF-36), the Hospital Anxiety and Depression Scale (HAD) and the Self-Esteem Scale (SES), Outcomes will be evaluated at baseline and at weeks 4, 8, 12, 16, 28, and 40, respectively. All adverse events that occur during this study will be recorded. If any participant withdraws from the trial, an intention-to-treat analysis (ITT) will be performed. CONCLUSION: This is a randomized, sham-controlled trial of CIA treatment for simple obesity. The results of this trial will provide more evidence on whether CIA is efficacious and safe for treating obesity. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT02936973. Registered on October 18, 2016.

aPKC inhibitors might be the sensitizer of chemotherapy and adoptive immunotherapy in the treatment of hASIPa-overexpressed breast cancer.

The atypical protein kinase C (aPKC) plays an important role in cell growth through the interaction with its substracts, including human ASIP (hASIP), which contains an aPKC phosphorylation site encoded by exon 17b of the gene. hASIP is expressed as numerous alternative splicing isoforms in the cells. Our results showed that hASIPa, an exon 17b-containing isoform of hASIP, is overexpressed in human breast cancer (HBC) MDA-MB-231, SK-BR-3, and MCF-7 cell lines and HBC specimens. The anticancer effects of 5-FU chemotherapy or adoptive immunotherapy and the synergic action of aPKC inhibitor against hASIPa-overexpressed HBC cells were tested. The results indicated that HBC MDA-MB-231 and SK-BR-3 cells were sensitive to 5-FU treatment in vitro. The combined treatment of aPKC inhibitor and 5-FU raised the anticancer activities against hASIPa-overexpressed HBC cells. The coculture of cytokine-induced killer (CIK) cells and autologous dendritic cells (DCs) with or without Her-2 peptide GP2 pulse created two new populations of effective immune-active T-cell populations called DC-modulated and cytokine-induced killer (DCIK) cells and peptide-DC-modulated and cytokine-induced killer (DCIK-P) cells. The DCIK cells showed cytotoxic activities on MDA-MB-231, SK-BR-3, and MCF-7 cells in MHC unrestricted manner. The DCIK-P cells possessed extra-enhanced cytotoxic activities against HLA-A2+/Her-2+ MDA-MB-231 cells in MHC restricted manner, but not for HLA-A2+/-/Her-2+ SK-BR-3 cells and HLA-A2+/Her-2+/- MCF-7 cells. The data suggested specific cytotoxic T-lymphocyte (CTL) activity of DCIK-P cells on MDA-MB-231 cells. The combined treatment of aPKC inhibitor with DCIK/DCIK-P cells further raised the anticancer activities against hASIPa-overexpressed HBC cells. The results demonstrated that the hASIPa/aPKC signaling pathway functions as an important regulator in the growth of HBC cells and aPKC inhibitor treatment showed the synergic activities on 5-FU or DCIK/DCIK-P cells adoptive immunotherapy against hASIPa-overexpressed HBC cells.

[Regularity of Acupoint Selection for Simple Obesity Treated by Acupoint Catgut Embedding Based on Complex Network Technology].

OBJECTIVE: To analyze the core acupoints and acupoint combinations of catgut embedding for simple obesity based on the complex network technology. METHODS: Articles about acupoint catgut embedding for simple obesity were collec-ted from databases of PubMed, CNKI, Wanfang and VIP from 1980 to 2016 by using keywords "simple obesity" "obesity" "acupoint embedding" "acupuncture" and "traditional Chinese medicine"， followed by constructing a database of acupoint prescription. Acupoint node (one node means an acupoint) weighted complex network was constructed by using complex network technique, followed by conducting centrality analysis and clustering analysis about the nodes using Matlab 2014, a software for revealing the core acupoint node and compatibility relations. At last, theresults (complex network diagram) were displayed using software Gephi 0.9.1. RESULTS: A total of 238 articles (all in Chinese) including 278 acupoints (of which 115 are meridian acupoints) were collected. The top 15 core acupoints are Tianshu (ST 25), Zusanli (ST 36), Zhongwan (CV 12), Fenglong (ST 40), San-yinjiao (SP 6), Quchi (LI 11), Yinlingquan (SP 9), Guanyuan (CV 4), Pishu (BL 20), Qihai (CV 6), Shenshu (BL 23), Shangjuxu (ST 37), Daheng (SP 15), Shuifen (CV 9), and Ganshu (BL 18), mainly distributing in the abdomen, lower limbs and back. Those acupoints with the highest core degree are attributed to the Stomach Meridian, Conception Vessel, Bladder Meri-dian, Spleen Meridian and Large Intestine Meridian. Regarding the compatibility of these acupoints, ST 25 and CV 12 have the highe-st correlation frequency, followed by ST 25 and ST 36, and ST 40 and ST 25, indicating the principle of different combinations being regional acupoints and meridian acupoints. CONCLUSION: In the treatment of simple obesity with catgut embedding, top 15 core acupoints as ST 25, ST 36, CV 12, ST 40, etc, and acupoint recipes as ST 25 and CV 12, ST 25 and ST 36, ST 40 and ST 25 are most frequently used in clinical practice.

Total parathyroidectomy combined with partial auto-transplantation for the treatment of secondary hyperparathyroidism.

BACKGROUND: Drug treatment for secondary hyperparathyroidism caused by chronic renal failure may be available at the early stage of the disease, but it is not as effective for serious patients. The aim of the study was to evaluate the effect of total parathyroidectomy combined with forearm autotransplantation in the uremic patients with secondary hyperparathyroidism. METHODS: From September 1999 through September 2006, parathroidectomy and autotransplantation was performed in 20 patients. The coherence between the results of preoperative parathyroid ultrasonography and surgical exploration were compared. The serum calcium concentration and intact parathyroid hormone (iPTH) were monitored preoperatively, intraoperatively, and postoperatively. RESULTS: A total of 71 hyperplastic parathyroid glands were resected in the 20 patients. The accordance rate of parathyroid localization between B-ultrasonography and intraoperative exploration was 94.4%. The average iPTH value was (110.90 +/- 67.42) ng/L, (433.80 +/- 243.72) ng/L, (48.80 +/- 42.69) ng/L, (229.04 +/- 172.68) ng/L and (232.39 +/- 224.05) ng/L at day 1, 2, 3, 7, 30 after operation respectively. The clinical symptoms were ameliorated and the levels of serum calcium concentration were controlled within the normal range after operation. Recurrent secondary hyperparathyroidism had happened in 1 case, 4 years postoperatively because of the development of autograft hyperplasia, and in another case 2 years postoperatively due to remnant of neck parathyroid glands. The clinical symptoms were all alleviated after re-operation. No surgical complication had occurred in any of the patients. CONCLUSIONS: The total parathyroidectomy with forearm autotransplantation is feasible, safe, and effective for patients with secondary hyperparathyroidism in the short term. The long-term effects should be further investigated.

CA IX is upregulated in CoCl2-induced hypoxia and associated with cell invasive potential and a poor prognosis of breast cancer.

Hypoxia, a common phenomenon during the development of malignant solid tumors including breast cancer, serves to propagate a cascade of molecular pathways triggered by hypoxia-inducible factor-1α (HIF-1α). Carbonic anhydrase IX (CA IX), one of the target genes of HIF-1α, has been reported to be involved in progression of malignant tumors. The objective of this study was to investigate the expression of HIF-1α and CA IX in hypoxia, involvement of CA IX in the regulation of migration and invasion/metastasis and its prognostic significance in breast cancer. We used cobalt chloride (CoCl2) as a hypoxia-mimetic agent and found that the expression of HIF-1α protein, CA IX mRNA and protein, is effectively upregulated, except for HIF-1α mRNA. Data showed that the elevated CA IX expression is closely related to in vitro cell migration and invasion, and the underlying mechanism of this process may be associated with epithelial-mesenchymal transition (EMT). The study of clinical tissue samples also demonstrated that CA IX is an independent prognostic marker that may serve as a useful clinical biomarker for predicting tumor progression and the invasion/metastasis of breast cancer. These results provide further insight into the role of CA IX in tumor progression and put forward further strong evidence as well as new consideration for CA IX target therapy.

MicroRNA-320a inhibits breast cancer metastasis by targeting metadherin.

Dysregulated microRNAs play important pathological roles in carcinogenesis that are yet to be fully elucidated. This study was performed to investigate the biological functions of microRNA-320a (miR-320a) in breast cancer and the underlying mechanisms. Function analyses for cell proliferation, cell cycle, and cell invasion/migration, were conducted after miR-320a silencing and overexpression. The specific target genes of miR-320a were predicted by TargetScan algorithm and then determined by dual luciferase reporter assay and rescue experiment. The relationship between miR-320a and its target genes was explored in human breast cancer tissues. We found that miR-320a overexpression could inhibit breast cancer invasion and migration abilities in vitro, while miR-320a silencing could enhance that. In addition, miR-320a could suppress activity of 3'-untranslated region luciferase of metadherin (MTDH), a potent oncogene. The rescue experiment revealed that MTDH was a functional target of miR-320a. Moreover, we found that MTDH was negatively correlated with miR-320a expression, and it was related to clinical outcomes of breast cancer. Further xenograft experiment also showed that miR-320a could inhibit breast cancer metastasis in vivo. Our findings clearly demonstrate that miR-320a suppresses breast cancer metastasis by directly inhibiting MTDH expression. The present study provides a new insight into anti-oncogenic roles of miR-320a and suggests that miR-320a/MTDH pathway is a putative therapeutic target in breast cancer.