Ras/Raf/MEK/ERK singal transduction plays an important role in cell proliferation, differentiation, apoptosis, metastasis and metabolism. This investigation focused on this signal pathway and chose farnesyl transferase (FTase) as the main target and Raf-1 kinase as the second target. A lot of compounds were selected to construct the pharmacophore models of farnesyl transferase inhibitors (FTIs) and Raf-1 kinase inhibitors by using computer-aided drug design (CADD). The pharmacophore of FTIs is constituted by a hydrogen bonding acceptor, an aromatic ring, a positive ionizable and two hydrophobic regions; the pharmacophore of Raf-1 kinase is constituted by a hydrogen donor, a hydrogen acceptor, a hydrophobic regions and an aromatic ring. There are some similarities between the two pharmacophores. After analysis of the constructions of these two pharmacophores, some new aminomethylbenzoic acid derivatives with good forecasting activity against both of FTase and Raf-1 kinase were designed with these new pharmacophore models.

Objective To apply the virtual reality in balance function training for Parkinson's Disease (PD). Methods From October, 2013 to January, 2016, 31 patients with balance disorders after PD were divided into two groups:the treatment group (n=17) received virtual reality balancing games training, while the control group (n=14) received routine balance training. They were assessed with Berg Balance Scale (BBS), Timed Up and Go Test (TUGT), static fall index (SFI), limits of stability (LOS), dynamic fall index (DFI) and modified Barth-el Index (MBI) before and four weeks after treatment. The frequency of falls in the next six weeks was recorded. Results The scores of BBS, TUGT, SFI, LOS, DFI and MBI improved significantly in both groups after treatment (P<0.05), and improved more in scores of BBS (t=2.095), TUGT (t=-2.091), DFI (t=-2.182), LOS scores (t=2.202) and MBI (t=3.036) in the treatment group than in the control group (P<0.05). There was positive correlation between scores of BBS and MBI (r=0.899, P<0.001). The frequency of falls was less in the treatment group than in the control group (P<0.05). Conclusion Virtual reality balance training can improve the balance function for PD.

To obtain active protein of pIL-18 expression in Lactococcus lactis, and to observe its biological activity, the total RNA was extracted as template from peripheral blood mononuclear cells. Porcine interleukin 18 (pIL-18) was amplified by RT-PCR. The resulting fragment was cloned into pAMJ399 L. lactis vector, and then transformed to L. lactis MG1363 cells by electroporation. Expression of pIL-18 protein was detected by SDS-PAGE and Western-blotting. Bioactivity of the product was tested by pig spleen lymphocyte proliferation test and cytopathogenic effect inhibition assay. The result of Western blotting and bioactivity test shows that the molecular weight of pIL-18 protein was 19 kDa. The react line was observed in both supernatant and precipitated of the recombinant bacteria pAMJ399-pIL18/MG1363. The expressed pIL-18 can promote the proliferation of pig spleen lymphocyte, and significantly inhibit virus multiplication. As conclusion, porcine interleukin-18 was successfully expressed in L. lactis, and the product was biologically active.
Animals ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Electroporation ; Genetic Vectors ; Interleukin-18 ; biosynthesis ; Lactococcus lactis ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Swine

The coding sequence for the mature peptide of porcine lactoferrin (Plf) was synthesized according to the codon usage of lactobacillus, to establish optimized porcine lactoferrin Lactobacillus expression system. The gene was ligated into the Xho I/BamH I site of Lactobacillus expression vector pPG612.1 and the recombinant plasmid pPG612.1-plf was transformed individually into Lactobacillus casei ATCC393, Lactobacillus pentosus KLDS1.0413, Lactobacillus plantarum KLDS1.0344 or Lactobacillus paracasei KLDS1.0652 by electroporation. After induction with xylose, expression of the recombinant proteins was detected by Western blotting and confocal laser scanning microscopy. Secretion of recombinant Plf proteins from four recombinant species was determined quantitatively by ELISA. The antibacterial activities of recombinant proteins were measured by agar diffusion method. The result shows that Plf was correctly expressed in four species of recombinant lactobacillus, with molecular weight of about 73 kDa. The expression levels in recombinant Lactobacillus casei, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paracasei were 9.6 μg/mL, 10.8 μg/mL, 12.5 μg/mL and 9.9 μg/mL, respectively. Antimicrobial activity experiment shows that the recombinant proteins were active against E. coli, Staphylococcus aureus, Salmonella typhimurium, Listeria, Pasteurella. The recombinant Plf expressed by recombinant Lactobacillus plantarum showed the best antibacterial activity among all recombinant lactobacillus species. These data represent a basis for the development and application of porcine lactoferrin from recombinant lactobacillus.
Animals ; Anti-Bacterial Agents ; biosynthesis ; Lactobacillus ; metabolism ; Lactoferrin ; biosynthesis ; Recombinant Proteins ; biosynthesis ; Swine

Occupational therapy can not only promote the development of motor and cognitive functions in children with developmental or other disabilities, but also improve their self-care ability, learning ability, participation skills and other activities.In 2019, the Aust Occup Ther J published a systematic review on the effects of occupational therapy interventions on disabled children, aiming to summarize the best evidence of occupational therapy and help occupational therapists and families to choose the effective treatment regimens.In this article, the systematic review was interpreted in light of the current status of domestic child occupational therapy, so as to allow clinicians to compare the pros and cons of different treatment methods and improve the efficacy.

To evaluate the immune responses of recombinant Lactobacillus casei 393 expressing Porcine Epidemic Diarrhea Viral (PEDV) N protein as oral vaccine, n gene of PEDV was subcloned into the expression vector pPG-1, and then transformed into L. casei 393 by electroporation, resulting in recombinant strain pPG-1-n/L, casei 393. The recombinant strains were induced to express interest protein, which was detected by Western blotting, immunofluorescence microscopy and the whole bacteria ELISA. And then BALB/C mice were used as an animal model immunized with recombinant strains by oral administration, and the immune efficacy was analyzed. The recombinant PEDV N protein showed the antigenic specificity, and was located on the bacterial cell walls of pPG-1-n transformed L. casei. The results of ELISA showed that the mice immunized with recombinant strains could produce remarkable special sIgA level in the feces, and high level of anti-PEDV N protein IgG in the serum (P < 0.01), but the induced antibodies in serum did not demonstrated neutralizing effect. Statistical significant difference was observed among the spleen lymphocyte proliferation index (LPI) among the immunization groups of mice and control groups. And there was significant increase. of IFN-gamma and IL-4 contents in the supernatant of spleen cell culture in immunized group. In conclusion, the oral immunizations with recombinant L. casei 393 can induce significant specific mucosal PEDV N-specific IgA response as well as serum IgG responses, and can evoke both mucosal immune and system immune responses.
Administration, Oral ; Animals ; Antibody Formation ; Coronavirus Infections ; prevention & control ; Female ; Immunity, Mucosal ; immunology ; Lactobacillus casei ; genetics ; metabolism ; Mice ; Mice, Inbred BALB C ; Nucleocapsid Proteins ; biosynthesis ; genetics ; immunology ; Porcine epidemic diarrhea virus ; immunology ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Swine ; Viral Vaccines ; administration & dosage ; immunology

Objective To investigate the relationship between total serum IgG concentration and tumor progression in patients with gastric cancer.Methods From January 2010 to May 2017,a total of 90 patients with gastric cancer in Sun Simiao Hospital of Beijing University of Chinese Medicine were enrolled as study subjects and 90 age-and-sex-matched adults from normal physical examination were enrolled as control subjects.The serum IgG subgroup and total IgG concentrations were measured by enzyme-linked immunosorbent assay.The relationships between serum IgG concentration in patients with gastric cancer and clinicopathologic features were analyzed.The numbers of plasma cells with CD138 antibody in both tumor and non-cancerous tissues were evaluated by immunohistochemistry.Results The total serum IgG concentration in patients with gastric cancer was significantly lower than that in healthy control group [(923.6±290.5) mg/dl vs.(1 072.5 ±298.6) mg/dl],with a significant difference (t =3.391,P =0.001).The total serum IgG and IgG1 concentrations were correlated with lymphatic metastasis (t =3.585,P ＜ 0.001;t =3.545,P ＜ 0.001) and gastric cancer stages (t =3.669,P ＜0.001;t =3.401,P ＜0.001).Furthermore,the CD138 + plasma cells in tumor tissues were significantly fewer than those in non-cancerous gastric mucosa (88.7 ± 31.9 vs.108.5 ± 33.4),with significant differences (t =2.778,P =0.007).Multivariate analysis suggested that total serum IgG concentration (HR =0.411,95％ CI:0.094-0.989,P =0.011) and lymph node metastasis (HR =3.148,95％ CI:1.988-5.297,P ＜ 0.001) were independent predictors for poor prognosis.Conclusion Decreased total serum IgG concentration is closely related to tumor progression and poor prognosis in patients with gastric cancer,which indicates that the impaired humoral immunity is associated with tumor progression.

Objective:To explore any effect of mechanical vibration on the expression of estrogen and brain-derived neurotrophic factor (BDNF) in ovariectomized rats with an osteoporotic fracture.Methods:Thirty 3-month-old female Wistar rats were divided randomly into a control group, an ovariectomy group and a vibration group, each of 10. Fractures were induced in the rats of all three groups. Twenty minutes of whole-body vertical vibration was applied to the vibration group at a frequency of 35Hz, 5 days a week for 6 weeks. After 2 and 6 weeks the fracture healing of each group was evaluated using X-rays, the levels of hippocampal estrogen were measured using enzyme-linked immunosorbent assays and fracture-end BDNF was quantified by immunoblotting.Results:After 2 and 6 weeks of vibration the average fracture healing in the vibration group was significantly greater than in the other 2 groups. The average estrogen content in the hippocampus of the vibration group was significantly higher than in the other 2 groups after both 2 and 6 weeks, while the average BDNF content in their fracture ends was significantly lower. The BDNF expression at the fracture end was significantly correlated with the fracture healing.Conclusion:Mechanical vibration can promote the expression of estrogen and BDNF in the hippocampus and accelerate fracture healing in osteoporotic rats.

Objective:To explore the abnormal brain metabolic pattern and connectivity in temporal lobe epilepsy (TLE) patients.Methods:18F-FDG PET images of 75 patients diagnosed as drug resistant unilateral TLE from January 2014 to December 2016 in Huashan Hospital of Fudan University were collected retrospectively, including 41 (22 males, 19 females, age (28.4±8.7) years) left TLE (LTLE) and 34 (13 males, 21 females, age (28.5±8.8) years) right TLE (RTLE). Forty-four healthy controls (24 males, 20 females, age (31.2±6.2) years) were also enrolled. The cerebral glucose metabolism in TLE patients and the controls were analyzed with statistical parametric mapping (SPM) 12. The brain connectivity based on glucose metabolism were analyzed with bilateral hippocampus and amygdala as seeds. Permutation test with 1 000 permutations was used to analyze data. Results:Compared to control group, in both LTLE and RTLE groups, hypometabolism was found in affected hippocampus, amygdala, insula and temporal gyrus and hypermetabolism was observed in health hippocampus, parahippocampal gyrus, amygdala, lenticular nucleus and thalamus. In addition, hypometabolism was also found in affected superior/middle frontal gyrus and hypermetabolism was also found in bilateral frontal-orbital gyrus, bilateral cerebellum, affected lenticular nucleus and thalamus in LTLE group. In both TLE groups, affected seeds exhibited increased connectivity with affected superior frontal gyrus, lingual gyrus, fusiform gyrus, superior/middle temporal gyrus and temporal pole (all P<0.05); affected seeds exhibited increased connectivity with health superior frontal gyrus ( P=0.005), lingual gyrus ( P=0.018) and transverse temporal gyrus ( P=0.016) in RTLE group in addition. Besides, affected seeds exhibited decreased connectivity with bilateral default mode network (DMN) (all P<0.05), affected caudate nucleus ( P=0.015) and health thalamus ( P=0.008), in a uniform distribution pattern in LTLE group, and with bilateral cerebral cortex in an irregular distribution pattern in RTLE group (all P<0.05). In LTLE group, health seeds exhibited more increased connections with superior ( P=0.005)/middle frontal gyrus ( P=0.042), health hippocampus ( P=0.038), parahippocampal gyrus ( P=0.019), amygdala ( P=0.038), posterior cingulate gyrus ( P=0.004), and bilateral fusiform gyrusand ( P=0.048) compared with RTLE group; while, in RTLE group, health seeds exhibited more decreased connections with health superior ( P=0.047), inferior frontal gyrus ( P<0.001), orbital frontal gyrus ( P<0.001) and rectus gyrus ( P=0.016) compared with LTLE group. Conclusion:Altered brain glucose metabolism and connectivity pattern are found and will elucidate the underlying metabolic pattern of TLE.