Objective To observe the relationship between the levels of plasma VEGF,PAI-1 and the severity of the diabetic retinopathy in type 2 diabetic patients.Methods ELISA method was used to measure the levels of VEGF and PAI-1.Results The level of VEGF was increased in all diabetic patients compared with the normal control group[(17.86?12.25)ng/L],and that of the patients with BDR[(93.41?54.69)ng/L]was higher than that of the patients with NDR[(52.17?21.81)ng/L]and PDR[(61.24?37.55)ng/L].The level of PAI-1 was also increased in all diabetic patients.The increase tendency was more obvious in patients with BDR[(58.29?20.53)?g/L]and PDR[(66.84?23.81)?g/L].Conclusion The damage of the vascular endothelial cell and the impaired fibrinolysis state exist in diabetic retinopathy.So it is very important to measure the levels of VEGF and PAI-1 for the early diagnosis and intervention treatment of diabetic retinopathy.

OBJECTIVE:To a evaluation method for the measurement uncertainty for the content of Bisacodyl enteric-coated tablet. METHODS:HPLC external standard method was conducted for content determination of Bisacodyl enteric-coated tablet, and mathematical model for uncertainty evaluation was established to systematically analyze and evaluate the influential factors in processes of solution preparation and instrument measurement. RESULTS:HPLC external standard method showed the content was 97.8%,confidence probability was 95%,expanded uncertainty was 2.8%,and determination result was (97.8 ± 2.8)%,k=2. CONCLUSIONS:The established method is suitable for the evaluation of measurement uncertainty for the content of Bisacodyl en-teric-coated tablet. Regularly calibrated verification for HPLC equipment and strict control of the weighing process will help to im-prove the accuracy measured by HPLC.

Natural products are important sources of drug discovery. However, the traditional methods of extraction and isolation, as well as chemical synthesis for obtaining natural products are associated with issues such as operational complexity, high costs, low efficiency, and environmental pollution. Constructing microbial cell factories through synthetic biology methods to produce medicinal natural products has the advantages of high efficiency, low cost, and environmental protection. Nevertheless, the scope and yield improvement of the products are limited by the limitations of enzymes in microbial cell factories. Protein engineering is considered one of the most effective approaches to overcome these limitations. This article introduces commonly used methods of protein engineering technology and summarizes its specific applications in improving enzyme performance, modifying the enzymatic environment, and promoting the development of synthetic biology tools in the field of pharmaceutical natural product synthesis. Furthermore, it analyzes the current bottlenecks and challenges in protein engineering and looks forward to its future application prospects, offering insights for the development and practical use of protein engineering technology.

Abortion, Induced ; adverse effects ; Adult ; Device Removal ; Embolization, Therapeutic ; Female ; Humans ; Intrauterine Devices ; Pregnancy ; Uterine Artery ; Uterine Rupture ; etiology

Objective The purpose of this study was to discuss the clinical significance of serum levels of Pro-gastrin-releasing peptide (ProGRP) in diagnosis,therapy monitoring and prognosis in patients with small cell lung cancer (SCLC).Methods Clinical diagnostic trial.Serum levels of ProGRP were measured by ELISA assays in 413 SCLC patients,418 NSCLC,120 with benign pulmonary diseases patients and 200 healthy subjects.Patients were recuited by the Shanxi Cancer Hospital from Dec.2005 to Oct.2008.Three hundreds and sixty-eight patients with SCLC were followed up from Dec.2005 to Oct.2011.The receiver operating characteristic curves (ROC) was used to set the cut-off value of ProGRP and the area under ROC (ROC-AUC).The sensitivity and specificity of ProGRP were analyzed for diagnosing SCLC.The survival analysis was performed by the Kaplan-Meier method and Cox's proportional hazards model for multivariate analysis of prognosis.Results Using healthy subjects group as control,the largest Youden index point of ROC was used to set the cut-off values of ProGRP and NSE (45.3 ng/L and 12.4 ng/L).The ROC-AUC of ProGRP was 0.798 (95％ CI:0.746-0.850)the sensitivity and specificity were 79.2％,98.1％ respectively.The AUC of NSE was 0.786(95％ CI:0.726-0.746),the sensitivity and specificity were 71.9％,96.7％ respectively; Combing detection of ProGRP and NSE,the sensitivity and specificity were 88.1％,95.8％ respectively.Serum levels of ProGRP in healthy subjects,benign pulmonary diseases,NSCLC and SCLC groups were 6.9 (5.3-8.6),36.8 (26.3-43.4),21.3 (18.6-35.2) and 1758.7 (368.4-2967.3) μg/L respectively.The serum levels of ProGRP in SCLC groups were significantly higher than those in the healthy group,benign pulmonary diseases group and NSCLC group (H =103.66,P =0.000).Serum levels of ProGRP in SCLC at stage Ⅰ-],stage m,stage Ⅳ were 543.3 (256.8-843.2),1440.6 (1042.4-2543.3) and 1897.6 (1586.5-3958.7) μg/L,respectively (H =25.974,P =0.000).Serum levels of ProGRP in 165 SCLC patients with complete remission(CR) were significantly declined after treatment (U =11.65,P ＜ 0.01).The levels of ProGRP in 146 SCLC patients with partial remission(PR) slowly decreased (U =9.17,P ＜ 0.01).Thirty-nine cases with progressive disease (PD)and 63 cases with stable disease(SD) presented elevated ProGRP levels (U =3.314,P ＜ 0.001 ; U =2.54,P ＜ 0.01,respectively).By the end of October 31st 2011,a total of 368 cases with SCLC were followedup.Ratio of follow-up was 89.1％.There were 56 deaths in 119 SCLC patients with ProGRP ＜ 1000 μg/L (median time =16.0 months,4-23 months) ; 159 deaths in 249 with ProGRP ＞ 1000 μg/L (median survival time =12.0 months,2-18 months).Median survival time of the two groups showed significant differences(x2 =11.04,P =0.001).Multivariate analysis by Cox's proportional hazards model revealed that ProGRP was independent prognostic factor related to the overall survival (OS) of SCLC patients.Conclusions The serum ProGRP is valuable tumor marker for diagnosis,treat monitoring and prognosis of SCLC.It's important to predict relapses and recurrence of diseases earlier,instruct therapy and prognosis assessment.

Objective Toevaluatethediagnosticvalueofhumanepididymisprotein4(HE4)andcar-bohydrate antigen 1 25 (CA1 25 )for distinguishing between benign and malignant ovarian neoplasms.Methods 1 1 09 patients with ovarian neoplasms were enrolled in this study,serum concentration of HE4 and CA1 25 was assayed using ELISA technique.And the markers were evaluated for significance separately and in combination. Results 1SerumlevelsofHE4andCA125weresignificantlyhigherinpost-menopausalwomenthanthosein pre-menopausal women(t=8.40,P<0.05;t=7.02,P<0.05).In addition,the more children the patients born,the higher serum levels of these two markers were(F=1 5.36,P<0.05;F=1 3.00,P<0.05).2 Serum HE4 levels were significantly higher in the ovarian cancer patients compared with those seen in patients with benign or borderline tumor(t=1 3.68,P<0.05;t=1 4.94,P<0.05).Serum CA1 25 levels were significantly higher in the ovarian cancer patients compared with those seen in patients with benign or borderline tumor(t=1 4.1 6,P<0.05;t=1 7.27,P<0.05).Morever,it also appared in the ovarian cancer patients with ascites and vascular embolism.Morever,the levels of HE4 were significantly higher in the ovarian cancer with ascites and vascular embolism than without it(t=7.08,P<0.05;t=4.41,P<0.05),the levels of CA125 were signifi-cantly higher in the ovarian cancer with ascites and vascular embolism than without it(t=9.67,P<0.05;t=4.75,P<0.05).3 During follow-up,serum HE4 and CA1 25 levels significantly decreased at 3 months after operation(t=9.86,P<0.05;t=5.12,P<0.05).4 Receiver operating characteristic curve,ROC)analysis revealed that no difference was observed in AUC values for HE4,CA1 25 and risk of ovarian malignancy algo-rithm(ROMA).5 Compared to CA1 25 ,HE4 had significantly higher specificity and lower sensitivity.Howev-er,sensitivity were increased when the two markers were combined with each other.However,the sensitivity of combination with two markers was higher than single detection and ROMA,but the specificity was lower in com-bination with two markers than single detection and ROMA.If we divide the ROMA by a woman′s menopausal status,ROMA has a higher sensitivity (73.84%,84.1 9%) and lower specificity (66.06%,66.67%). Conclusions ThelevelsofCA125hasahighsensitivity,andthelevelsofHE4isahighspecificity.CA125 combined with HE4 can provide a more sensitivity and accurate predictor of ovarian cancer than either alone.

Objective To investigate the low-dose hyper-radiosensitivity (HRS)/induced radioresistance (IRR) in A549 cells synchronized at G2 phase and the role of ATM kinase in the process.Methods Human lung adenocarcinoma cell line A549 was synchronized at G2 phase by aphidicolin.The ATM-specific activator and inhibitor,chloroquine and KU55933,were used to regulate the activity of ATM.The colony formation assay was used to evaluate cell survival.Flow cytometry was used to determine the cell cycle of radiation-exposed A549 cells synchronized at G2 phase.Immunofluorescence was used to observe the dynamics of γ-H2AX fluorescence and evaluate the efficiency of DNA repair in A549 cells synchronized at G2 phase.Western blot was used to detect the expression of phosphorylated ATM (Ser1981) and ATM.Results A549 cells synchronized at G2 phase had substantially enhanced HRS than non-synchronized cells.The dose-induced transition from HRS to 1RR was in accordance with the dose-response pattern of early G2/M checkpoint.However,with the same threshold dose,the activation of early checkpoint occurred earlier and lasted longer than normal.The activation of ATM kinase inhibited HRS and enhanced DNA repair,while the inhibition of ATM kinase enhanced HRS and hindered DNA repair.Conclusions ATM kinase-mediated early G2+M checkpoint is a molecular switch for HRS in synchronized A549 cells.Low-dose irradiation with G2-phase synchronization and ATM inhibitor can enhance the low-dose radiosensitivity.

BACKGROUND:Occlusion of the middle cerebral artery trunk can cause large infarctions.Clinically,thrombolytic therapy,defibrillation,vasodilation,and surgery are often used to treat these large infarctions.However,the therapeutic efficacy of these therapeutic interventions in promoting prognosis remains controversial.Nalmefene,an opioid receptor antagonist,exhibits a neuroprotective effect.Few clinical studies are reported on the therapeutic efficacy of nalmefene in the treatment of large cerebral infarctions.OBJECTIVE:To investigate the therapeutic efficacy of nalmefene in restoring the neurologic function of patients with middle cerebral artery trunk infarction in comparison with conventional treatment as a control.METHODS:A total of 236 patients with middle cerebral artery trunk infarction who will receive treatment at the First Hospital of Jilin University,China will be randomly divided into a control group (n=116) and a nalmefene group (n=120).Patients in the control group will receive conventional treatment.Patients in the nalmefene group will receive 10 successive days of intravenous nalmefene hydrochloride injection based on conventional treatment.The primary outcome of this study is the effective rate at 20 days (10 days after treatment).The secondary outcomes of this study include (1) the National Institutes of Health Stroke Scale (NIHSS) score at 20 days,used to evaluate neurologic function deficits;(2) Glasgow Coma Scale score at 0 (before treatment) and 10 days;(3) serum level of matrix metalloproteinase 9 at 0,5 and 10 days (before treatment and 5 and 10 days of treatment);(4) MRI perfusion imaging of the head at 0 and 10 days.This trial was ragistered with the Chinese Clinical Trial Registry (registration number:ChiCTR-IOR-17013871).RESULTS AND CONCLUSION:Preliminary experiment results ravealed that compared with the control group,NIHSS score was signficiantly decreased,effective rate was increased,Glasgow Coma Scale score was significantly increased,serum level of matrix metalloproteinase 9 was significantly decreased,cerebral blood flow and cerebral blood volume on the lesion side were signficiantly increased,and the mean transit time of contrast agent on the lesion side was significantly shortened in the nalmefene group.Findings from this study will provide clinical evidence for use of nalmefene in combination with conventional treatment for large cerebral infarctions and provide data support that this combined therapy can improve the prognosis in patients with large cerebral infarctions.

Objective To establish a reproducible mouse model of hepatic veno-occlusive disease (HVOD) after allogeneic bone marrow transplantation (aallo-ABMT) and explore its pathogenesis.Methods Balb/c mice were randomly divided into three groups:(1) normal saline (NS) control group; (2) total body irradiation (TBI) group; (3) allogeneic bone marrow transplantation (allo-BMT) group.Liver weight,total bilirubin (TBil),tumor necrosis factor α (TNF-a),interleukin 6 (IL-6) and monocyte chemotactic protein 1 (MCP-1) were detected on the day 0,5,10,15 and 20 after transplantation.Hepatic vein and sinusoid congestion,infiltration of inflanmatory cells,and damage to hepatic cells and vascular endothelial cells were observed under the light microscopy after HE staining.Fibrosis of hepatic sinusoids and venule was observed under the light microscopy after Masson staining.Results Liver weight and TBil levels were elevated at 5th day and reached the peak at 15th day after all-ABMT.The changes of hepatic congestion and edema were obviously observed and there was infiltration of inflammatory cells at 5th and 10th day after alloABMT.At 15th and 20th day,hepatic congestion,edema and necrosis were reduced and liver damage was mainly presented with liver fibrosis and inflammatory infiltration.All mice died within 10 days after TBI,and hepatic congestion and edema were aggravated.As compared with NS control group,TNF-α,IL-6 and MCP-1 concentrations were significantly increased after all-ABMT.Conclusion A reproducible mouse model of hepatic veno-occlusive disease after all-ABMT was successfully established,and the pathogenesis was closely related to endothelial damage caused by total body irradiation,inflammatory cell infiltration and increased concentrations of cytokines.

Lipase-mediated biodiesel production becomes increasingly important because of mild reaction conditions, pollution free during the process and easy product separation. Compared with traditional immobilized lipase, whole cell biocatalyst is promising for biodiesel production because it is easy to prepare and has higher enzyme activity recovery. Rhizopus oryzae IFO4697 can be used as the catalyst for biodiesel production. To further study the stability of the whole cell biocatalyst is extremely important for its further application on large scale. This paper focuses on the stability study of Rhizopus oryzae IFO4697 when used for the methanolysis of renewable oils for biodiesel production. The results showed that water content was important for achieving high catalytic activity and good stability of the biocatalyst. The optimum water content was found to be 5%-15%. Both particle size and desiccation methods showed no obvious effect on the stability of the biocatalyst. With GA cross-linking pretreatment, the stability of the biocatalyst could be improved significantly. When Rhizopus oryzae IFO4697 repeatedly used for next batch reaction, direct vacuum filtration was found to be a good way for the maintenance of good stability of the biocatalyst. Under the optimum reaction conditions, the methyl ester yield could keep over 80% during 20 repeated reaction batches.
Biocatalysis ; Bioelectric Energy Sources ; microbiology ; Biofuels ; Cells, Immobilized ; metabolism ; Lipase ; metabolism ; Rhizopus ; metabolism ; Soybean Oil ; metabolism