Abnormal distribution of laminin α2 chain in glomerular basement membrane in membranous nephropathy / 临床与实验病理学杂志

To study the abnormal distribution of type Ⅳ collagen and laminin chains in glomerular basement membrane (GBM) in membranous nephropathy (MN).Methods 52 cases of MN were collected and staged according to electron microscopic morphological characteristics,and 10 cases of kidney tissues of minimal change disease were used as normal GBM control.Distribution pattern of or5 (Ⅳ) chain,laminin α5and β2 chains,and laminin α2 and β1 chains were detected using immunofluorescence method.Results In minimal change disease,α5 (Ⅳ) chain,laminin α5 and β2 chains all showed continuously linear positive expression along GBM,and laminin α2 and β1 chains were negatively expressed in GBM.In stage Ⅰ MN,α5 (Ⅳ) chain,laminin α.5 and β2 chains all showed continuous linear positive expression along GBM.In stage ⅡMN,the expression of α5 (Ⅳ) chain was increased and showed abundant spikes on the basis of continuous linear positive staining along GBM,and the expression of laminin α5 and β2chains was increased,and segmental spikes were seen on the basis of continuous linear positive staining along GBM.In stage ⅢMN,the expression of α5 (Ⅳ),laminin α5 and β2 chains was also enhanced and segmental double tracks were seen.The expression of laminin α2 chain was negative in GBM in stage ⅠMN,but granular positive expression along GBM was seen in stage Ⅱ and stage Ⅲ MN.No positive expression of laminin β1chain was seen in GBM in different stages in MN.Conclusion The GBM thickness in MN originates not only from intrinsic type Ⅳ collagen chains and laminin chains,but also from laminin α2chain,which only exist in glomerulus mesangium in normal condition.

Multi-step purifications of botulinum neurotoxin A light chain and identification of its metalloproteases activity / 军事医学

Objective To obtain highly purified botulinum neurotoxin A light chain(BoNT-ALC) protein in E.coli by genetic engineering and multi-step purifications, and identify its metalloproteases activity.Methods The full-length of BoNT-ALC was cloned from BoNT A by PCR and inserted into plasmid pET-22b.Then pET-22b-ALC was transformed into E.coli BL21( DE3) strains and induced by IPTG.The protein was purified by Ni-NTA sepharose,anion exchange column and gel filtration.The enzymatic activity of the protein was identified by SNAP-25.Results and Conclusion A highly purified and homogeneous protein is obtained, which shows good enzymatic activity.

Study on Diuretic Effect ofMori Cortex Aqueous Extracts and Chemical Split Fractions / 世界科学技术-中医药现代化

This study was aimed to investigate the diuretic effect ofMori Cortex and to identify effective fractions inM. Cortex. Metabolic cages were used to firstly observe the diuretic effect ofM. Cortex aqueous extracts on rats. Medications were continuously given for 5 days to screen for the best dosage of diuretic effect. Picric acid assays were used to detect creatinine levels in serum and urine after 5-day medication. Then, the diuretic effect of each chemical split fraction inM. Cortex was studied in order to indentify the effective parts. The results showed that after administration ofM. Cortex aqueous extracts for 3 to 5 days, compared with the control group, there was a significant diuretic effect on rats (P<0.05 orP<0.01). And the medium-dose ofM. Cortex aqueous extracts showed the best effect (P<0.01). However,M. Cortex aqueous extracts had no significant effect on creatinine levels in serum and urine. Assays of diuretic effect of chemical split fractions ofM.Cortex indicated that compared with the control group, 30% ethanol fraction and fatty oil fraction had the best diuretic effect (P<0.01). It was concluded thatM.Cortex aqueous extracts had a significant diuretic effect. And the chemical fractions contributed mostly to this effect were mainly existed in the 30% ethanol fraction and fatty oil fraction.