Many major neurodegenerative diseases are associated with proteins misfolding and aggregation, which are also called "neurodegenerative conformational disease". The interaction of gene mutation and environmental factors are probably primary events resulting in oligomer and aggregate formations of proteins. Moreover, the dysfunctions of protein control systems, i.e. the ubiquitin-proteasome system and autophagy-lysosomal system, also contribute to the neurodegenerative process. The present review mainly summarizes protein misfolding and aggregation in the development of neurodegenerative conformational disease and the underling mechanisms, as well as upregulation of heatshock proteins as a promising treatment method for this kind of disease.

BACKGROUND: Changing the local environment of spinal injury promotes the repair and regeneration of injured nerve and recovery of partial nervous function of spinal cord. Transplantation of olfactory ensheathing cells can improve the local internal environment of injured spinal cord.OBJECTIVE: To probe into the effect and safety of transplantation of olfactory ensheathing cells on functional repair of spinal cord and nerve in patients with obsolete spinal injury DESIGN: Self-control experiment.SETTING: Wards of the Department of Surgery, Taian Rongjun Hospital of Shandong Province.PARTICIPANTS: Totally 48 patients admitted for obsolete spinal injury in the Department of Surgery, Taian Rongjun Hospital, between June 2004 and July 2005 were recruited. There were 39 males and 9 females, aged 7 to 59 years with the mean of 36 years.METHODS: ①Cell culture: Olfactory bulb of aborted fetus was digested into single olfactory ensheathing cells, which were then cultured and puri fied for 1 to 2 weeks, and finally made into single cell suspension. ②Operation and cell transplantation: Under general anesthesia, the purified single cell suspension (about 0.05-0.20 mL) of olfactory ensheathing cells was injected into the corresponding spinal injury site through multiple points with home-made syringe of 0.45 mm in diameter. Stitches were taken out at postoperative 10 to 14 days. ③Evaluation of spinal function: Injury Scoring Standard made by American Spinal Injury Association (ASIA) was used for scoring, comparison and statistical analysis at postoperative 1 day and 2 weeks to 2 months. ④Spinal function of 48 patients was observed or followed up through telephone at postoperative 3 weeks to 1 year.MAIN OUTCOME MEASURES: Changes of postoperative sensory function of the patients. Changes of postoperative motor function of the patients. Changes of postoperative automatic nervous system of the patients.RESULTS: ①All the 48 patients had improvement in spinal function, and continued improved tendency was found in the observation and follow-up through telephone at postoperative 3 weeks to 1 year. ②Scoring by ASIA for sensory function was higher after operation than before operation (touch sensation: 56.9, 51.2, P ＜ 0.01; pain sensation: 55.2, 48.3, P ＜ 0.01). Sensory function was improved obviously at the lower shift of sensory level,generally more than 2 segments. ③Scoring by ASIA for motor function was higher after operation than before operation (44.8, 40.7, P ＜ 0.01), but the improvement was slow. ④Scoring by ASIA for automatic nervous system was higher after operation than before operation (18.0, 14.5, P ＜ 0.01); diaphoresis, increased enterokinesia and other automatic nervous system improved earliest.CONCLUSION: Transplantation of olfactory ensheathing cells promotes the spinal and neurofunctional recovery of patients with malignant spinal injury, and the therapeutic method is safe.

Purpose To investigate the value of T2WI mild-moderate signal and restricted diffusion in the context of liver imaging reporting and data system (LI-RADS) (2014 edition) in the diagnosis of hepatocellular carcinoma (HCC) with cirrhosis caused by hepatitis B virus.Materials and Methods A total of 77 lesions (LI-RADS 3-5,size of 1.1 cm×0.7 cm-12.7 cm×9.1 cm) of 69 HCC patients in Beijing Friendship Hospital from January 2012 to November 2016 were retrospectively analyzed.All these patients underwent MRI scan and multiphase dynamic enhanced scan.The images were analyzed by two radiologists.If a disagreement occurred,liver accelerated volume acquisition and multiphase dynamic enhanced scan were combined to reach a consensus.The contrast noise ratio (CNR) and apparent diffusion coefficient (ADC) of T2WI and diffusion weighted imaging (DWI) sequences were compared,as well as the identification of the two signs.Results There was no statistically significant difference between T2WI mild-moderate signal and restricted diffusion in the identification of lesions (LI-RADS 3-5) (P＞0.05),while the sensitivity with DWI b=0 (61.0％) was significantly lower than DWI b=600 s/mm2 (70.1％) (P＜0.05).The CNR of all DWI sequences (b=0,600 s/mm2) were larger than those of T2WI (P＜0.01).The ADC of small lesions (diameter ＜2 cm) were larger than those of larger lesions (diameter ＞2 cm) [(1.57+0.37)×10-3 mm2/s vs.(1.37+0.51)×10 3 mm2/s,P＜0.05].Conclusion There is no significant difference in sensitivity of lesions between T2WI mild-moderate signal and restricted diffusion.However,due to different CNRs,DWI with b=600 s/mm2 is more obvious for the lesions,and can be first investigated in practice.

Objective:To analyze the safety and clinical efficacy of invasive treatment for portal vein thrombosis after splenectomy or devascularization.Methods:Invasive treatment was retrospectively analyzed from Jan 2016 to Jan 2020. In 319 cases who met the inclusion criteria.Result:There were complications in 41 cases and no death;The average portal vein pressure before and after thrombus clearance treatment was (25.6±4.9) mmHg and (14.7±4.1) mmHg respectively ( t=2.53, P<0.05); Thrombus decreased significantly in most patients. Conclusion:Invasive therapy is a safe and effective method for patients complicated with portal vein thrombosis after splenectomy or devascularization.

Objective:To investigate the acute and long-term toxicity of GJ-4 extracted from Gardenia jasminoides J.Ellis, and to provide safety basis for its development as a new drug for the treatment of dementia. Methods:In the acute toxicity experiment, 30 ICR mice were randomly divided into control group, gardenia extract 2.5 g/kg group and gardenia extract 5.0 g/kg group, 10 mice in each group. The mice in the 2.5 g/kg and 5.0 g/kg gardenia extract groups were administrated with GJ-4 suspension. The control group was given 0.5% sodium carboxymethyl cellulose (CMC-Na) by gavage. The mice were given continuous gavage for 7 days. The mortality, body weight and general condition of mice were recorded. The levels of ALT, ALP, BUN and creatinine (CRE) in serum were measured by automatic biochemical detector. In the long-term toxicity experiment, 75 ICR mice were divided into control group and gardenia extract 100, 250, 500, 1 000 mg/kg group according to the random number table method, 15 mice in each group. The GJ-4 suspension of Gardenia extract 100, 250, 500 and 1 000 mg/kg were administrated to the stomach respectively in the gardenia extract 100, 250, 500 and 1 000 mg/kg groups, and 0.5% CMC-Na of the same volume was administrated to the stomach in the control group once a day for 30 days. The mortality, weight and mental state of mice were recorded. The organ index and the levels of ALT, ALP and BUN in serum were observed.Results:In the acute toxicity experiment, the mental state and diet of mice in each group were good, and there was no death within 7 days. Compared with the control group, there was no significant differences in body weight, heart index, liver index and kidney index between the two groups ( P>0.05). Compared with the control group, the level of BUN (10.17 ± 0.82 mmol/L vs. 11.25 ± 0.47 mmol/L) in the gardenia extract 2.5 g/kg group significantly decreased ( P<0.05), and the level of ALP (116.0 ± 10.75 U/L vs. 148.0 ± 25.73 U/L) in the gardenia extract 5.0 g/kg group significantly decreased ( P<0.05). In the long-term toxicity experiment, the mice were in good mental state and had good diet, and no death occurred. Compared with the control group, there was no significant differences in body weight, heart index, kidney index, spleen index and serum ALT, ALP and BUN levels between the two groups ( P>0.05). The liver index (4.9 ± 0.56 vs. 4.38 ± 0.49) in the 250 mg/kg gardenia extract group significantly increased ( P<0.01), and the thymus index (0.09 ± 0.02 vs. 0.14 ± 0.04) significantly decreased ( P<0.05). Conclusions:The Gardenia jasminoides extract GJ-4 has no obvious toxicity in acute and long-term toxicity experiment, indicating that GJ-4 is safe.

OBJECTIVE: To investigate whether the Runx2 gene can induce the differentiation of human amniotic mesenchymal stem cells (hAMSCs) to ligament fibroblasts in vitro and promote the tendon-bone healing in rabbits. METHODS: hAMSCs were isolated from the placentas voluntarily donated from healthy parturients and passaged, and then identified by flow cytometric identification. Adenoviral vectors carrying Runx2 gene (Ad-Runx2) and empty vector adenovirus (Ad-NC) were constructed and viral titer assay; then, the 3rd generation hAMSCs were transfected with Ad-Runx2 (Ad-Runx2 group) or Ad-NC (Ad-NC group). The real-time fluorescence quantitative PCR and Western blot were used to detect Runx2 gene and protein expression to verify the effectiveness of Ad-Runx2 transfection of hAMSCs; and at 3 and 7 days after transfection, real-time fluorescence quantitative PCR was further used to detect the expressions of ligament fibroblast-related genes [vascular endothelial growth factor (VEGF), collagen type Ⅰ, Fibronectin, and Tenascin-C]. The hAMSCs were used as a blank control group. The hAMSCs, hAMSCs transfected with Ad-NC, and hAMSCs were mixed with Matrigel according to the ratio of 1 : 1 and 1 : 2 to construct the cell-scaffold compound. Cell proliferation was detected by cell counting kit 8 (CCK-8) assay, and the corresponding cell-scaffold compound with better proliferation were taken for subsequent animal experiments. Twelve New Zealand white rabbits were randomly divided into 4 groups of sham operation group (Sham group), anterior cruciate ligament reconstruction group (ACLR group), anterior cruciate ligament reconstruction+hAMSCs transfected with Ad-NC-scaffold compound group (Ad-NC group), and anterior cruciate ligament reconstruction+hAMSCs transfected with Ad-Runx2-scaffold compound group (Ad-Runx2 group), with 3 rabbits in each group. After preparing the ACL reconstruction model, the Ad-NC group and the Ad-Runx2 group injected the optimal hAMSCs-Matrigel compunds into the bone channel correspondingly. The samples were taken for gross, histological (HE staining and sirius red staining), and immunofluorescence staining observation at 1 month after operation to evaluate the inflammatory cell infiltration as well as collagen and Tenascin-C content in the ligament tissues. RESULTS: Flow cytometric identification of the isolated cells conformed to the phenotypic characteristics of MSCs. The Runx2 gene was successfully transfected into hAMSCs. Compared with the Ad-NC group, the relative expressions of VEGF and collagen type Ⅰ genes in the Ad-Runx2 group significantly increased at 3 and 7 days after transfection ( P<0.05), Fibronectin significantly increased at 3 days ( P<0.05), and Tenascin-C significantly increased at 3 days and decreased at 7 days ( P<0.05). CCK-8 detection showed that there was no significant difference ( P>0.05) in the cell proliferation between groups and between different time points after mixed culture of two ratios. So the cell-scaffold compound constructed in the ratio of 1∶1 was selected for subsequent experiments. Animal experiments showed that at 1 month after operation, the continuity of the grafted tendon was complete in all groups; HE staining showed that the tissue repair in the Ad-Runx2 group was better and there were fewer inflammatory cells when compared with the ACLR group and the Ad-NC group; sirius red staining and immunofluorescence staining showed that the Ad-Runx2 group had more collagen typeⅠ and Ⅲ fibers, tending to form a normal ACL structure. However, the fluorescence intensity of Tenascin-C protein was weakening when compared to the ACLR and Ad-NC groups. CONCLUSION Runx2 gene transfection of hAMSCs induces directed differentiation to ligament fibroblasts and promotes tendon-bone healing in reconstructed anterior cruciate ligament in rabbits.
Pregnancy ; Female ; Humans ; Rabbits ; Animals ; Vascular Endothelial Growth Factor A/metabolism* ; Fibronectins/metabolism* ; Collagen Type I/genetics* ; Tenascin/metabolism* ; Collagen/metabolism* ; Anterior Cruciate Ligament/surgery* ; Mesenchymal Stem Cells ; Tendons/metabolism* ; Fibroblasts/metabolism*

Endophytic fungi are promising producers of bioactive small molecules. Bioinformatic analysis of the genome of an endophytic fungus