The structural gene encoding mature peptide of extracellular ? amylase was amplified from the genome DNA of hyperthermophilic archaeon Pyrococcus furiosus by PCR The recombinant plasmid pUC19 amy was constructed by inserting the amplified segment into vector pUC19 The recombinant vector pYX212 amy was constructed by ligate the heterogeneous fragment of pUC19 amy into the multiple cloning site of pYX212, an expression vector of yeast Saccharomyces cerevisiae W303 A1 were transformed with pYX212 amy by electroporation The transformant expressed the activity of the thermophilic ? amylase successfully The recombinant enzyme has the similar enzymatic properties as the extracellular ? amylase produced by Pyrococcus furiosus : it shows an enzymatic activity optimum at pH 5 0, and its optimal temperature for enzymatic activity is about 90℃, more than 50% of its initial enzymatic activity is still detectable after it was incubated at 121℃ for 30 minutes

Objective To study the risk factors of intracranial hemorrhage after endovascular therapy of cere-bral arteriovenous malformations embolization.Methods the clinical records and images of 230 patients with cAVMs were retrospectively reviewed.Patients with intracranial hemorrhage after endovascular therapy in 3 days were recorded and the risk factors of hemorrthage by using multivariate logistic regression for guiding the treatment strategy were studied.Results There were 15 patients occurred intracerebral hemorrhages.After treatment the area of cAVMs was significantly smaller than that before treatment[(2.18 ±0.91)cm vs (6.67 ±1.56)cm],the difference lvas a statis-tically significant (t =9.627,P <0.05).A partial AVM reduction of ≥36.5%(P =0.031)and hypertension(P =0.025)were considered as risk factors for hemorrhage after endovascular therapy of cAVWs.Conclusion Patients receives a partial AVMreduction of ≥36.5% or with hypertension history have a tendancy of increasing hemorrhage after endovascular therapy of cAVMs.It's helpful for preventing intracerebral hemorrhage if the arterial blood pressure maintains after endovascular therapy.

The production of 2, 3-butanediol and succinic acid by a moderate halophile under anaerobic condition was investigated. This halophile, termed Salinivibrio YS, was isolated from the solid samples collected from Aydingkol Lake. Based on the single factor experiment, the parameters and their values for the production were obtained. Then, the optimum values of these parameters by the orthogonal experiments were obtained: temperature, 33 degrees C; initial pH of fermentation, 8.0; the pH during fermentation, 7.0; the concentration of acetic acid was 3 g/L and NaC1 was 10 g/L. Finally, a 3-L fermentation based on these conditions was carried out. After 108 h of fermentation under anaerobic condition, 35.05 g/L of 2, 3-butanediol and 22.46 g/L of succinic acid were obtained. About 50% of total glucose conversion was achieved. The study on 2, 3-butanediol and succinic acid by a halophile under anaerobic condition will expand the applications of halophiles and open a new area of production of 2, 3-butanediol and succinic acid.
Anaerobiosis ; Butylene Glycols ; metabolism ; Fermentation ; Halobacteriales ; metabolism ; Succinic Acid ; metabolism ; Vibrionaceae ; metabolism

AIM:To study the role of amifostine on the formation of benzo [a]pyrene (BaP)-induced abdomi-nal aortic aneurysm ( AAA) in C57BL/6J mice and the underlying mechanism .METHODS: RAW246.7 mononuclear macrophage in vitro were divided into control group , DMSO group, BaP group, low dose (1 μmol/L) amfostine treated group, middle dose (5 μmol/L) amfostine treated group and high dose (25μmol/L) amfostine treated group .The influ-ence of BaP on the expression of matrix metalloproteinase (MMP)-9, MMP-12, TNF-α, NF-κB in the RAW246.7 mono-nuclear macrophages in vitro was determined by Western blot .Male C57BL/6J mice (8 months old) were divided into con-trol group, model group (AngII+BaP group), low dose (50 mg/kg) amfostine treated group and high dose (100 mg/kg) amfostine treated group.After 6 weeks, the abdominal aorta were isolated .The aortic tissues were subjected to HE and Masson staining.The vascular wall structure , infiltration of macrophage , the expression of MMP-9, MMP-12, TNF-α, NF-κB were evaluated by Western blot and immunochemistry staining .RESULTS:Amifostine attenuated BaP-induced expres-sion of TNF-α, MMP-9, MMP-12, NF-κB in the RAW246.7 mononuclear macrophages (P<0.05).The results of animal experiments showed that the incidence of AAA in high dose amifostine treated group were significantly lower than that in low dose amifostine treated group and model group (P<0.05).Immunohistochemistry staining observation showed that amifos-tine inhibited the aortic macrophage infiltration more obviously in high amifostine treated group compared with model group and low dose amifostine treated group (P<0.05).Compared with model group and low dose amifostine treated group , the MMP-9, MMP-12, TNF-αand NF-κB expression of abdominal aorta in high amifostine treated group was reduced signifi -cantly ( P<0.05 ) .CONCLUSION: Amifostine inhibits BaP-induced activation of macrophages , and also prevents the formation of abdominal aortic aneurysm in C 57BL/6J mice induced by BaP by inhibition of the NF-κB pathway, macro-phage infiltration and the expression of TNF-αand MMPs.

Objective To investigate the diagnostic value of serum human epididymis protein 4 (HE4) ,carbohydrate antigen 125 (CA125) ,carbohydrate antigen antigen (CA199) ,carbohydrate antigen 153 (CA153) and alpha fetoprotein (AFP) in the early di-agnosis of ovarian cancer .Methods From February 2014 to October 2016 ,117 patients with ovarian cancers who were treated in this hospital were selected ,including 69 cases of ovarian cancer and 48 cases of benign ovarian lesions ,and 70 healthy volunteers were selected as control group .The serum levels of HE4 ,CA125 ,CA199 ,CA153 and AFP were measured in all subjects . Results The positive rates of HE4 ,CA125 ,CA199 ,CA153 and AFP in the ovarian cancer group were 59 .42% ,68 .12% ,33 .33% , 46 .38% and 39 .13% ,respectively ,which were significantly higher than those in the benign ovarian lesion group and the control group (P<0 .05) .The sensitivity of CA125 in the diagnosis of ovarian cancer was 68 .11% ,the specificity was 88 .98% ,the nega-tive predictive value was 78 .33% ,the positive predictive value was 82 .68% ,Youden index was 0 .571 .The diagnostic efficiency was better than that of other tumor markers .Pathological examination revealed 34 cases of serous adenocarcinoma ,18 cases of mucinous adenocarcinoma and 17 cases of endometrioid carcinoma in 69 cases of ovarian cancer .The positive rate of serous adenocarcinoma CA125 was 85 .29% ,significantly higher than mucinous carcinoma and endometrioid carcinoma (χ2 =9 .398 ,P<0 .05) .Conclusion CA125 has a good application value in the early diagnosis of ovarian cancer ,the positive rate is higher in serous adenocarcinoma .

Objective To explore the value of integrin αvβ3 micro positron emission tomography (microPET)/CT imaging with 68Ga-DOTA-RGD2 in diagnosis of breast cancer osteolytic bone metastases.Methods MDA-MB-231 was injected in left ventricle of hairless mice at the density of 1.0×10 7/mL.Animal model with parathyroid hormone(PTH)-induced osteolysis in the calvarium was established.Animals were randomly divided into RGD group (n=10) and FDG group (n=10), and were detected by 68Ga-DOTA-RGD2microPET/CT and 18F-NaFmicroPET/CT respectively.Animals were sacrificed and bone lesions were collected for pathological examination.Results Bone radiotracer uptake ratio of osteolytie lesion to normal calvrium (O/N) was (4.18±0.570) by 68Ga-DOTA-RGD2, which was significantly higher than (1.24±0.28) in group FDG by 18F-NaFmicroPET/CT imaging (P<0.05).Conclusion 68Ga-DOTA-RGD2 integrin receptor αvβ3 imaging can effectively detect breast cancer bone metastases and locate osteolytic lesions.

Objective To explore the value of integrin αvβ3 micro positron emission tomography (microPET)/CT imaging with 68Ga-DOTA-RGD2 in diagnosis of breast cancer osteolytic bone metastases.Methods MDA-MB-231 was injected in left ventricle of hairless mice at the density of 1.0×10 7/mL.Animal model with parathyroid hormone(PTH)-induced osteolysis in the calvarium was established.Animals were randomly divided into RGD group (n=10) and FDG group (n=10), and were detected by 68Ga-DOTA-RGD2microPET/CT and 18F-NaFmicroPET/CT respectively.Animals were sacrificed and bone lesions were collected for pathological examination.Results Bone radiotracer uptake ratio of osteolytie lesion to normal calvrium (O/N) was (4.18±0.570) by 68Ga-DOTA-RGD2, which was significantly higher than (1.24±0.28) in group FDG by 18F-NaFmicroPET/CT imaging (P<0.05).Conclusion 68Ga-DOTA-RGD2 integrin receptor αvβ3 imaging can effectively detect breast cancer bone metastases and locate osteolytic lesions.

Mycoplasma pneumoniae pneumonia (MPP) complicated with cerebral venous sinus thrombosis (CVST) is rare.We retrospectively analyzed the clinical data of two patients with refractory mycoplasma pneumoniae pneumonia (RMPP) complicated with CVST who were hospitalized in Xi′an children′s Hospital from December 2018 to April 2019, inquired the relevant literature, analyzed the clinical diagnosis and treatment characteristics, and discussed the diagnosis and treatment measures of RMPP complicated with CVST.Two cases were 6-year-old girls with fever and cough as the main symptoms.After physical examination, the respiratory sounds of the affected lung decreased, and the sounds of phlegm and dampness could be heard in both lungs.Mycoplasma pneumoniae (MP) antibody and RNA were positive.Chest CT showed lobar pneumonia with a large number of pleural effusion.The effect of macrolide antibiotics anti infection treatment was not good.Headache symptoms occurred during the course of the disease, and serum D-dimer increased significantly.Brain MRI showed CVST, including 1 case with lower extremity pain, and B-ultrasound showed right lower extremity arterial embolism.After anti infection, thrombectomy, anticoagulation and symptomatic treatment, 2 cases were discharged.When children with MPP, especially those with RMPP, have extracranial thrombosis and/or neurological symptoms, accompanied by elevated serum D-dimer, the possibility of CVST should be considered, and brain MRI examination should be performed in time to confirm and actively treat, which can reduce or avoid the occurrence of sequelae.Thrombosis may be related to excessive inflammatory reaction and vascular endothelial injury caused by MP infection.

Autophagy is a highly conservative cellular self-protective behavior dependent on lysosomes, and can be used as an important factor in promoting or preventing cancer, and its effect is related to the type and development of tumors. A full understanding of autophagy pathway is helpful to improve the diagnosis and treatment of tumors. Studies have shown that autophagy is closely related to the occurrence and development of oral tumors. Autophagy-related genes and signal pathways play a dual regulatory role on oral tumors. This article reviews the latest progress in the regulatory mechanism and therapeutic effect of autophagy on oral tumors.

Objective:To investigate the accurate diagnosis and differential diagnosis of non-primary solid malignant tumors in breast needle core biopsy.Methods:Twenty-three cases of breast, axilla or neck lymph nodes pathologically diagnosed as non-primary solid malignant tumors were collected at the First Affiliated Hospital of Nanjing Medical University, Nanjing, China from January 2013 to March 2023. The differential diagnoses and diagnostic features were analyzed, based on combining clinical data, histology, and expression characteristics of biomarkers.Results:All patients were female, with age ranging from 29 to 75 years (average 56 years). The average time from the diagnosis of primary tumor to the current diagnosis was 21 months (0 to 204 months).The primary sites included the ovary (9 cases), the lung (5 cases), the gastrointestinal tract (4 cases), the pancreas, intrahepatic bile duct, thyroid gland, nasal cavity and forearm skin (1 case each). No carcinoma in situ was found in any of the cases. The morphological differences were significant among the tumors, but similar to the primary tumors. The tumors of neuroendocrine and female reproductive tract had great morphological and immunophenotypic overlaps with breast cancer. Metastatic lung cancer cells showed obvious atypia and tumor giant cells. The morphology and immunophenotype of metastatic serous carcinoma of female reproductive system might resemble invasive micropapillary carcinoma of the breast. Metastatic adenocarcinoma of the gastrointestinal tract often had features of mucous secretion. Metastatic neuroendocrine tumors were bland in appearance and morphologically similar to solid papillary carcinoma of breast, but negative for ER. TRPS1 was mostly negative (18/23) and variably positive in ovarian (4/9) and intrahepatic bile duct (1/1) tumors.Conclusions:The diagnosis of breast needle core biopsy specimen should be combined with clinical history, imaging study, and careful examination of histological features, such as presence of in situ component, morphological similarity between the primary and metastatic tumors, and using appropriate markers to differentiate the primary from metastatic tumors.