BACKGROUND:The Guilu Erxian Glue consists of Testudinis Plastrum,Cornu Cervi,Lycii Fructus,and Ginseng Radix.In earlier clinical observations,it is discovered that using Guilu Erxian Glue to treat patients with liver-kidney deficiency type knee osteoarthritis effectively alleviated knee pain,increased the range of motion,and improved walking ability.However,the exact mechanism by which oral administration of Guilu Erxian Glue can produce local therapeutic effects on the knee joint is still unclear.OBJECTIVE:To investigate the effects of Guilu Erxian Glue on gut microbiota in rats with knee osteoarthritis and to evaluate its mechanism using 16S rDNA sequencing and machine learning analysis.METHODS:Totally 18 female SD rats were randomly divided into three groups:blank group,model group,and Guilu Erxian Glue group,with 6 rats in each group.A knee osteoarthritis model was prepared using the destabilization of the medial meniscus surgical method.After successful modeling,the Guilu Erxian Glue group was given a decoction of Guilu Erxian Glue by gavage,while the blank and model groups were given an equal amount of distilled water.After 28 days of continuous intervention,high performance liquid chromatography was used to detect the active ingredients of Guilu Erxian Glue.MRI imaging was used to observe the condition of rat knee articular cartilage.Fecal samples were collected;DNA was extracted using a kit,amplified and purified by PCR,and an Illumina sequencing library was constructed.The Illumina MiSeq platform was used for high-throughput sequencing to generate raw sequence data.After obtaining the raw data,QIIME2 software was used to process the data.Linear Discriminant Analysis Effect Size analysis and random forest algorithm were used to screen for differential species in microbial data.KEGG and MetaCyc functional pathway analyses were used to explore the association between key microbial communities and experimental groups.Linear discriminant analysis effect values and random forest algorithm were used to screen for differential species.Association networks were used to analyze the interactions between microbial communities,and machine learning methods were used to analyze the composition and changes of gut microbiota.RESULTS AND CONCLUSION:(1)LC-MS component identification was conducted on the traditional Chinese medicine formula of Guilu Erxian Glue,and a total of 7 effective ingredients were identified.(2)MRI imaging showed that synovitis scope of high-density shadows in rats of the Guilu Erxian Glue group was reduced,and the degeneration of medial femoral condyle cartilage was less than that in the model group.(3)16S rDNA sequencing showed that the model group rats exhibited significant microbial imbalance,with a significant decrease in the abundance of Firmicutes and Bacteroidetes at the phylum level,while the proportion of Proteobacteria increased significantly(P＜0.05).The gut microbiota structure of rats in the Guilu Erxian Glue group was significantly improved,and the proportion of Firmicutes and Bacteroidetes increased,restoring a more diverse microbiota composition,approaching that of the blank group(P＜0.05).(4)KEGG and MetaCyc functional pathway analysis showed that the Guilu Erxian Glue group significantly activated multiple metabolic pathways,including amino acid metabolism,lipid metabolism,and biotin synthesis pathways(P＜0.05).(5)The results indicate that Guilu Erxian Glue contains seven active ingredients,and the changes in gut microbiota of knee osteoarthritis rats were analyzed using 16S rDNA sequencing.Guilu Erxian Glue can significantly improve the imbalance of gut microbiota,restore the abundance of beneficial bacteria,and have a significant impact on the composition of gut microbiota,providing scientific basis for the efficacy and mechanism of Guilu Erxian Glue.

BACKGROUND:The Guilu Erxian Glue consists of Testudinis Plastrum,Cornu Cervi,Lycii Fructus,and Ginseng Radix.In earlier clinical observations,it is discovered that using Guilu Erxian Glue to treat patients with liver-kidney deficiency type knee osteoarthritis effectively alleviated knee pain,increased the range of motion,and improved walking ability.However,the exact mechanism by which oral administration of Guilu Erxian Glue can produce local therapeutic effects on the knee joint is still unclear.OBJECTIVE:To investigate the effects of Guilu Erxian Glue on gut microbiota in rats with knee osteoarthritis and to evaluate its mechanism using 16S rDNA sequencing and machine learning analysis.METHODS:Totally 18 female SD rats were randomly divided into three groups:blank group,model group,and Guilu Erxian Glue group,with 6 rats in each group.A knee osteoarthritis model was prepared using the destabilization of the medial meniscus surgical method.After successful modeling,the Guilu Erxian Glue group was given a decoction of Guilu Erxian Glue by gavage,while the blank and model groups were given an equal amount of distilled water.After 28 days of continuous intervention,high performance liquid chromatography was used to detect the active ingredients of Guilu Erxian Glue.MRI imaging was used to observe the condition of rat knee articular cartilage.Fecal samples were collected;DNA was extracted using a kit,amplified and purified by PCR,and an Illumina sequencing library was constructed.The Illumina MiSeq platform was used for high-throughput sequencing to generate raw sequence data.After obtaining the raw data,QIIME2 software was used to process the data.Linear Discriminant Analysis Effect Size analysis and random forest algorithm were used to screen for differential species in microbial data.KEGG and MetaCyc functional pathway analyses were used to explore the association between key microbial communities and experimental groups.Linear discriminant analysis effect values and random forest algorithm were used to screen for differential species.Association networks were used to analyze the interactions between microbial communities,and machine learning methods were used to analyze the composition and changes of gut microbiota.RESULTS AND CONCLUSION:(1)LC-MS component identification was conducted on the traditional Chinese medicine formula of Guilu Erxian Glue,and a total of 7 effective ingredients were identified.(2)MRI imaging showed that synovitis scope of high-density shadows in rats of the Guilu Erxian Glue group was reduced,and the degeneration of medial femoral condyle cartilage was less than that in the model group.(3)16S rDNA sequencing showed that the model group rats exhibited significant microbial imbalance,with a significant decrease in the abundance of Firmicutes and Bacteroidetes at the phylum level,while the proportion of Proteobacteria increased significantly(P＜0.05).The gut microbiota structure of rats in the Guilu Erxian Glue group was significantly improved,and the proportion of Firmicutes and Bacteroidetes increased,restoring a more diverse microbiota composition,approaching that of the blank group(P＜0.05).(4)KEGG and MetaCyc functional pathway analysis showed that the Guilu Erxian Glue group significantly activated multiple metabolic pathways,including amino acid metabolism,lipid metabolism,and biotin synthesis pathways(P＜0.05).(5)The results indicate that Guilu Erxian Glue contains seven active ingredients,and the changes in gut microbiota of knee osteoarthritis rats were analyzed using 16S rDNA sequencing.Guilu Erxian Glue can significantly improve the imbalance of gut microbiota,restore the abundance of beneficial bacteria,and have a significant impact on the composition of gut microbiota,providing scientific basis for the efficacy and mechanism of Guilu Erxian Glue.

Objective:To investigate clinical significance of the detection of bone mineral density(BMD)and serum levels of vitamin D in elderly patients with Parkinson's disease(PD).Methods:Sixty elderly patients with PD(the PD group)admitted in our hospital from June 2016 to December 2018 were enrolled in this retrospective study.And 60 healthy persons confirmed by annual health check-up matched for age and sex during the same period were included as the control group.PD patients were divided into the osteoporosis group(n=23)and the non-osteoporosis group(n=37). The clinical data, bone mineral density and serum vitamin D level were compared between the two groups.Multivariate Logistic regression method was used to analyze related factors for osteoporosis in PD patients.Results:The incidences of osteoporosis and vitamin D deficiency were higher in PD group than in control group[23 cases(38.3%) vs.13 cases(21.7%)、35 cases(58.3%) vs.21 cases(35.0%), all P<0.05]. Bone mineral density and serum 25-(OH)D level were lower in PD group than in control group[(0.77±0.08)g/m 2vs.(0.83±0.09)g/m 2, (25.65±8.65)nmol/L vs.(39.80±10.74)nmol/L, t=4.381 and 8.439, P<0.05]. The age, course of disease and H-Y grade were higher and serum level of 25-(OH)D was lower in the osteoporosis group than in the control group( P<0.05). Spearman correlation analysis showed that BMD and 25-(OH)D were negatively correlated with age, course of disease and H-Y stage, respectively, and BMD was positively correlated with 25-(OH)D( r=0.396, P<0.05). Multivariate Logistic regression analysis showed that vitamin D deficiency was an independent risk factor for osteoporosis in elderly PD patients( OR=2.332, 95% CI: 1.772-8.224, P<0.01). Conclusions:The incidence of osteoporosis is high in elderly PD patients, and vitamin D deficiency is often present.Vitamin D deficiency may be an independent risk factor for osteoporosis.

Objective To investigate the effects of human umbilical cord mesenchymal stem cells (hUCMSCs)transplantation on the treatment of hypoxic-ischemic encephalopathy(HIE). Methods A total of 25 HIE patients were randomly divided into stem cell transplantation group(15 case)and control group(10 cases). The patients in transplantation group were given intravenous infusion of hUCMSCs,which isolated under sterile condition in vitro and cultured, while in control group were treated with routine drug treatment. Neurological function( American National Institutes of Health Stroke Scale( NIHSS ),Barthel index (BI)),extrapyramidal function(Unified Parkinson's disease questionnaire(UPDRS)),cognition and emotional reaction(The mini mental state examination(MMSE),the 14 item Hamilton Depression Scale(HAMD14)and HAMD24)were all assessed before and after transplantation for 14 d,90 d and 180 d respectively to evaluate the clinical efficacy of hUCMSCs transplantation. Results There was no significant difference between two groups in terms of each function before transplantation. The scores of transplantation group were all obviously improved after treatment for 14 d,90 d and 180 d compared to that of before treatment,and the therapy effect in transplantation group was significantly better than that of the control group( NIHSS:Ftime =4. 372,P=0. 031;Ftime*group =4. 175,P=0. 038;Fgroup =3. 897,P=0. 045.BI:Ftime =4. 728,P=0. 044;Ftime*group =4. 894,P=0. 037;Fgroup =4. 284,P=0. 039.UPDRS:Ftime =5. 112,P=0. 047;Ftime*group =4. 895,P=0. 045;Fgroup=3. 879,P =0. 031.MMSE:Ftime =5. 135,P =0. 039;Ftime*group =3. 213,P =0. 036;Fgroup =4. 184,P=0. 045.HAMD14:Ftime =3. 977,P =0. 049;Ftime*group =4. 587,P =0. 038;Fgroup =4. 381,P =0. 041.HAMD24:Ftime =3. 845,P =0. 033;Ftime*group =4. 125,P=0. 035;Fgroup =3. 547,P=0. 034). Conclusion Transplantation of hUCMSCs is safe and effective for treatment of HIE,which can significantly improve the neurological function,extrapyramidal function,cognition and emotion.

BACKGROUND:Currently, neural stem celltransplantation can be performed through three main approaches:local lesions, blood circulation, and cerebrospinal fluid.
OBJECTIVE:To review the transplantation of neural stem cells or neural precursor cells via the cerebrospinal fluid in the treatment of central nervous system diseases.
METHODS:A computer-based search of PubMed and CHKD databases was performed to retrieve articles concerning transplantation of neural stem cells via the cerebrospinal fluid, and its application and therapeutic mechanism in the treatment of central nervous system diseases in both animal experiment and clinic study published from 2000 to 2009.
RESULTS AND CONCLUSION:It is suitable for neural stem cellsurvival, proliferation, and differentiation in the cerebrospinal fluid. Transplantation of neural stem cells via the cerebrospinal fluid is effective and feasible to treat central nervous system diseases. However, some problems have not been solved, such as the source of neural stem cells, the optimal time window and celldose, the safety and the long-term effect. Further studies are needed to pave the way for the intrathecal injection of neural stem cells in the treatment of central nervous system diseases.