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 investigate the association of Tim-1 protein expression and its gene polymorphism with nutritional parameters in patients with systemic lupus erythematosus (SLE).Methods Peripheral blood samples were collected from 126 patients with SLE.Serum Tim-1 protein levels were detected by ELISA,and the Tim-1 gene-416G＞C,-1454G＞A polymorphism was detected by PCR-RFLP.Prealbumin,ceruloplasmin,and retinol conjugated protein levels were determined by immunoturbidimetry.Ferritin and 1,25-dihydroxy vitamin D3 levels were detected by electrochemiluminescence.Results Concentrations of serum Tim-1 protein,prealbumin,ceruloplasmin,retinol binding protein,ferritin,and 1,25-dihydroxy vitamin D3 were 249.7±30.2 pg/mL,226±42 μg/mL,363±95 μg/mL,29.4± 13.2 μg/mL,355± 164 ng/mL,and 26.4-± 11.5 ng/mL,respectively.In the-416G＞C site,GG,GC,and CC genotypes accounted for 11.9％,57.1％,and 31.0％,respectively.In the-1454G＞A site,GG,GA,and AA genotypes accounted for 67.5％,26.2％,and 6.3％,respectively.The Tim-1 protein concentration did not differ significantly between the different genotypes of the-416G＞C site (F=0.575,P=0.564) or-1454G＞A site (F=1.255,P=0.289).Tim-1 level was significandy negatively correlated with prealbumin (r =-0.176,P =0.033),and positively correlated with ceruloplasmin (r =0.205,P =0.014) and 1,25-dihydroxy vitamin D3 (r=0.166,P=0.042).The serum prealbumin level decreased significantly (P=0.027) in patients harboring the GG genotype in the-1454G＞A site,whereas the serum 1,25-dihydroxy vitamin D3 level decreased significandy (P =0.024) in patients with the AA genotype in the-1454G＞A site.Conclusion Serum Tim-1 protein level and the-1454G＞A polymorphism of Tim-1 gene are associated with the nutritional function of patients with SLE.

To investigate the expression of CD151 in human atherosclerosed artery and explore its clinical implications, Western blot and immunohistochemical techniques were used to determine the protein expression of CD151 in arterial tissues with atherosclerosis taken from 36 patients, including 26 cases who received bypass operation for peripheral artery atherosclerosis and 6 cases who died from coronary heart disease. The expression of CD151 in normal artery tissues from 15 healthy organ donators were also measured to serve as control. The results showed that expression of CD151 protein in atherosclerotic arteries was significantly higher than that in normal artery. In ath erosclerotic arteries, CD151 expression was localized in vascular smooth muscle cells (VSMCs) in all tunica media and in partial subintima, while in normal artery, sparse expression was found in tunica media near adventitia. It is concluded that high CD151 protein expression in artery is associated with atherosclerosis and CD151 plays an important role in the atherosclerosis related to VSMC. The expression of CD151 in human atherosclerotic artery depends on the extent of atherosclerotic dam age, it's independent of risk factors.

Traumatic cerebrospinal fluid leakage commonly presents in traumatic brain injury patients, and it may lead to complications such as meningitis, ventriculitis, brain abscess, subdural hematoma or tension pneumocephalus. When misdiagnosed or inappropriately treated, traumatic cerebrospinal fluid leakage may result in severe complications and may be life-threatening. Some traumatic cerebrospinal fluid leakage has concealed manifestations and is prone to misdiagnosis. Due to different sites and mechanisms of trauma and degree of cerebrospinal fluid leak, treatments for traumatic cerebrospinal fluid leakage varies greatly. Hence, the Craniocerebral Trauma Professional Group of Neurosurgery Branch of Chinese Medical Association and the Neurological Injury Professional Group of Trauma Branch of Chinese Medical Association organized relevant experts to formulate the " Chinese expert consensus on the diagnosis and treatment of traumatic cerebrospinal fluid leakage in adults ( version 2023)" based on existing clinical evidence and experience. The consensus consisted of 16 recommendations, covering the leakage diagnosis, localization, treatments, and intracranial infection prevention, so as to standardize the diagnosis and treatment of traumatic cerebrospinal fluid leakage and improve the overall prognosis of the patients.