Objective: To prepare chitosan neurotoxin nanoparticles (CS-NT-NP) and study its effect on the permeability of blood brain barrier and the serum levels of S100β by intranasal administration. Methods: A formamide extraction-ultraviolet spectrophotometry method was employed to determine the concentration of Evans blue (EB) in brain by different routes of administration and preparations. Qualitative analysis of fluorescence intensity and distribution of EB in brain tissue was performed by fluorescence microscopy. The serum S100β protein concentration was determined by ELISA. Results: Compared with the control group and NT group, CS-NT-NP could significantly increase the content of EB in the brain with time-effect relation and reached the peak at 120 min (P < 0.01); Compared with muscle injection and ip injection, intranasal administration could significantly increase the content of EB and reached peak time quickly. The results were consistent with the experimental results of qualitative analysis of fluorescence intensity and distribution of EB in brain tissue by fluorescence microscopy and S100β protein in serum. It was consistent with the experimental results of S100β protein. Conclusion: CS-NT-NP by intranasal administration can significantly increase the permeability of BBB, and further increase the drug concentration in the brain, which is a good carrier of macromolecular drugs into the brain.

In clinic, some urinary protein makers can dynamically and noninvasively reflect the degree of renal tubular injury in patients with diabetic nephropathy (DN). These urinary biomarkers of tubular damage are broadly divided into two categories. One is newfound, including kidney injury molecule-1 (Kim-1), neutrophil getatinase-associated lipocalin (NGAL), liver-type fatty acid-binding protein (L-FABP) and cystatin C (CysC); the other one is classical, including beta2 microglobulin (beta2-MG), retinal binding protein (RBP) and N-acetyl-beta-D-glucosaminidase (NAG). It is reported that, the increases in urinary protein markers are not only closely related to the damage of tubular epithelial cells in DN patients, but also can be ameliorated by the treatment with Chinese herbal compound preparations or Chinese herbal medicine. Recently, although urinary proteomics are used in the protein separation and identification, the traditional associated detection of urinary protein markers is more practical in clinic. At present, it is possible that the associated detection of urinary biomarkers of glomerular and tubular damages may be a feasible measure to reveal the clinical significance of urinary protein markers in DN patients and the interventional effects of Chinese herbal medicine.
Biomarkers ; urine ; Diabetic Nephropathies ; complications ; drug therapy ; urine ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Medicine, Chinese Traditional ; methods ; Proteinuria ; complications

To analyze the characteristic of urinary protein spectrum in patients with stage III diabetic nephropathy (DN) and its compliance with traditional Chinese medicine (TCM)symptom, for the sake of providing a basis for clarifying the rules of TCM syndrome differentiation in DN. Adopting the traditional epidemiological retrospective method, thirty-eight TCM syndromes and urinary protein with medium or low molecular weight, as well as urinary enzyme, including 24 h urinary protein (Upro), urinary albumin( UAlb), urinary retinal binding protein( URBP), urinary cystatin C (UCysC), urinary N-acetyl-beta-D-glucosaminidase (UNAG), were collected from 108 patients with stage III DN, and a multiple factor regression analysis between them was conducted. As the results, the levels of Upro, UAlb, URBP, UCysC, and UNAG were increased in 108 patients with stage III DN. Qi-Yin deficiency type was the major type. The level of UAlb in patients with Qi-Yin deficiency type was significantly higher than those without Qi-Yin deficiency type (P < 0.05). The elevation of Upro with the factors as swift digestion with rapid hungering, lassitude and lack of strength, weakness of waist and knees was complied, the elevation of UA1b with the factors as dry mouth with desire to drink, the elevation of URBP with the factors as numbness of extremities, shortness of breath, the elevation of UCysC with the factors as clear urine in large amounts, and the elevation of UNAG with the factors as frequent micturition, were complied respectively. In conclusion, for 108 stage III DN patients. The increase in urinary protein spectrum including UAlb, URBP, UCysC, and UNAG is the major characteristic. Shen and Pi are the major organs related to the appearance of urinary protein; Pi-Shen deficiency is the basic pathogenesis. The level of UAlb is taken as one of the objective syndrome factors for Qi-Yin deficiency type. The levels of UNAG and UCysC are possibly the objective syndrome factors for Shen-Qi deficiency type.
Diabetic Nephropathies ; complications ; diagnosis ; urine ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; methods ; Middle Aged ; Proteinuria ; complications ; urine ; Qi ; Regression Analysis ; Yin-Yang

OBJECTIVE: To determine the regulation effect of bone morphogenetic protein-4 (BMP-4) on the proliferation and differentiation of rat hepatic precursor cells. METHODS: We used Noggin (200 ng/mL) as the function blocking control of BMP-4, and the hepatic precursor cells of WB-F344 were treated with recombinant BMP-4 at 50 ng/mL at different time points. The proliferation of WB-F344 cells were tested by methyl thiazolyl tetrazolium (MTT) colorimetric assay. The ultrastructural characters of differentiated WB-F344 cells regulated by BMP-4 were observed under a transmission electron microscope. RT-PCR was used to examine mRNA expression of specific molecular markers for different cellular phenotypes potentially differentiated from the WB-F344 cells. RESULTS: At different time points, the absorbance values in the BMP-4 treatment groups were higher than those in the control groups of Noggin and blank treatment (P<0.01). The WB-F344 cells treated with BMP-4 exhibited typical ultrastructural characters of well-differentiated epithelial cells. The hepatocyte mRNA markers were more significantly promoted in the differentiated WB-F344 cells in the BMP-4 treatment group than those in the other 2 control groups. CONCLUSION BMP-4 can promote the proliferation and directional differentiation towards hepatocytes of rat hepatic precursor cells of WB-F344.
Animals ; Bone Morphogenetic Protein 4 ; antagonists & inhibitors ; genetics ; physiology ; Carrier Proteins ; pharmacology ; Cell Differentiation ; Cell Line ; Cell Proliferation ; Hepatocytes ; cytology ; Rats ; Recombinant Proteins ; Stem Cells ; cytology

OBJECTIVE: To determine the treatment effects of transplanted hepatic progenitor cells (WB-F344 cells) combined with heparin on the acute liver injury in SD rats. METHODS: A total of 2*10(7) hepatic stem cells (WB-F344) infected with GFP lentivirus and 8 μL heparin were transplanted through the spleen in SD rats with acute liver injury, which was induced by an intraperitoneal injection of CCl4. The liver and spleen tissues underwent fluorescence examination 1 day after the transplantation. The liver functions were tested, and the liver tissues were histopathologically examined on the 3rd, 7th, 14th, and 28th day of the cell transplantation. RESULTS: The transfected WB-F344 cells expressed GFP 3 days after the lentivirus infection and were found in the rat liver 1 day after the WB-F344 transplantation. The liver function and histopathological recovery of the liver tissues in the group of WB-F344 transplantation were better than those of the control group (P<0.05). CONCLUSION Transplantation of hepatic stem cells combined with heparin can promote the liver recovery in rats with acute liver injury induced by CCl4.
Animals ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; Heparin ; therapeutic use ; Hepatocytes ; transplantation ; Liver Failure, Acute ; chemically induced ; therapy ; Male ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation ; methods

OBJECTIVETo investigate the anti-motion sickness efficacy and influence on the blood level of some hormones of a Chinese prescription composed of 10 herbs such as spina date seed.

METHODSAccording to the report by Cramptom and Lucot, SD rats and Beagle dogs were rotated around a horizontal axis, and the rat behavior of pica for Kaolin and the latency to vomit in dog were observed. In addition, guinea pigs were rotated around a vertical axis, and the nystagmus was recorded. Blood levels of corticosterone, adrenocorticotrophic hormone (ACTH), corticotropin releasing hormone (CRH) and arginine vasopressin (AVP) in rats were measured with radioimmunoassay. The influences of the extracted mixture of herbs on these variables were simultaneously investigated.

RESULTSCompared with control group, oral administration of the extracted mixture of herbs: (1) significantly inhibited the rat behavior of pica for Kaolin and prolonged the latency to vomit in dog dose-dependently; (2) decreased the frequency of nystagmus and mean slow phase speed in rat; (3) reduced the elevation of corticosterone, ACTH, CRH and AVP in rat blood induced by rotatory stimulation; and (4) these effects of the extracted mixture of herbs were almost identical to dimenhydrinate.

CONCLUSION(1) The extracted mixture of Chinese Medicinal Herbs we used could inhibit motion sickness effectively. (2) This drug could reduce the blood levels of hormones of hypothalamic-pituitary-adrenocortical axis and AVP elevated by provocative rotatory stimulation.

Adrenocorticotropic Hormone ; blood ; Animals ; Arginine Vasopressin ; blood ; Corticosterone ; blood ; Corticotropin-Releasing Hormone ; blood ; Dogs ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Guinea Pigs ; Male ; Motion Sickness ; blood ; drug therapy ; Phytotherapy ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the association between genetic variations in methylenetetrahydrofolate reductase (MTHFR) and the risk of colorectal cancer in a Chinese population.

METHODSOne hundred and ninety-eight cases of colorectal cancer and 420 controls were genotyped for detecting MTHFR C677T and A1298C polymorphisms by PCR-RFLP methods. The adjusted odds ratios (OR) and 95% confidence intervals (CI) were calculated by using unconditional logistic regression model.

RESULTSThe frequency of the MTHFR C677T allele found among cases was significantly more frequent than that among controls (51.3% to 44.4%, P = 0.02). As compared with the 677CC genotype, the 677TT genotypes had a 67% (95% CI, 1.05 - 2.66) increased risk of colorectal cancers. However, there appeared no any correlations between the genetic variations of MTHFR and the cancer metastasis.

CONCLUSIONThese findings demonstrated that the genetic variation of MTHFR C677T should be a genetic susceptibility factor for colorectal cancer in a Chinese population.

Asian Continental Ancestry Group ; genetics ; China ; Colorectal Neoplasms ; ethnology ; genetics ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Logistic Models ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Odds Ratio ; Polymerase Chain Reaction ; Polymorphism, Restriction Fragment Length ; Polymorphism, Single Nucleotide ; Risk Factors

OBJECTIVETo construct a recombinant adenovirus of survivin vector and provid valuable reference for gene therapy of laryngeal cancer.

METHODSThe survivin gene was cloned by PCR. After confirmation by enzyme restriction analysis and sequencing, the gene and the adenovirus vector were recombined together to construct the recombinant adenovirus vector. The recombinant adenovirus vector was confirmed via both sequencing and digestion restriction analysis, and then linearized and transfected into the HEK 293 cell line to generate recombinant adenovirus.

RESULTSThe sequence analysis demonstrated that the survivin gene sequence was the same as published in the literature, suggesting that a recombinant adenovirus vector has been successfully constructed.

CONCLUSIONSA survivin recombinant adenovirus has been successfully constructed.

Adenoviridae ; genetics ; Genetic Vectors ; HEK293 Cells ; Humans ; Inhibitor of Apoptosis Proteins ; genetics ; metabolism ; Plasmids ; Polymerase Chain Reaction ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transfection

Objective:To investigate the predictive value of controlling nutritional status (CONUT) score in the prognosis of patients with advanced diffuse large B-cell lymphoma (DLBCL).Methods:A retrospective case series study was performed. The clinical data of 654 patients newly diagnosed with advanced DLBCL diagnosed in 7 medical centers in Huaihai Lymphoma Working Group from October 2009 to January 2022 were retrospectively collected. All the patients received rituximab-based immune chemotherapy regimens. The patients were randomly assigned to the training set (458 cases) and the validation set (196 cases) in a 7:3 ratio. The clinicopathological data of patients were collected, and the CONUT score was calculated based on albumin, lymphocyte count, and total cholesterol. The optimal critical value of CONUT scote was determined by using MaxStat method. Kaplan-Meier method was used to draw survival curves; Cox proportional hazards model was used to make univariate analysis and multivariate analysis on the factors influencing overall survival (OS). The efficacy of CONUT score in combination with the International prognostic index (IPI) and an enhanced IPI (NCCN-IPI) in predicting OS was evaluated by using receiver operating characteristic (ROC) curves.Results:The median follow-up time of 654 patients was 38.1 months (95% CI: 35.3 months- 40.9 months), and the 5-year OS rate was 49.2%. According to the MaxStat method, the optimal critical value for CONUT score was determined to be 6 points. All the patients were classified into the normal nutritional status group (CONUT score ≤ 6 points, 489 cases) and the poor nutritional status group (CONUT score > 6 points, 165 cases). The results of the multivariate analysis showed that CONUT score > 6 points, male, lactate dehydrogenase >240 U/L, high white blood cell count, low hemoglobin level and age > 60 years were independent risk factors for OS of patients with advanced DLBCL (all P < 0.05). Patients in the poor nutritional status group (CONUT score > 6 points) had worse OS compared with that in the normal nutritional status group in the overall cohort of advanced DLBCL. Subgroup analysis revealed that among patients with Eastern Cooperative Oncology Group-performance status (ECOG PS) score < 2 points, IPI low-intermediate risk, IPI intermediate-high risk, NCCN-IPI low-intermediate risk, and NCCN-IPI intermediate-high risk, the patients in the poor nutritional status group (CONUT score > 6 points) had worse OS compared with that in the normal nutritional status group (CONUT score ≤ 6 points) (all P < 0.05). Conclusions:CONUT score has a certain value in the assessment of the prognosis of patients with advanced DLBCL, and its predictive efficacy is further improved when combined with IPI and NCCN-IPI.

Objective To evaluate the effect of an emollient containing Prinsepia utilis Royle oil extracts and other extracts on clinical symptoms and disease recurrence in children aged 2-12 years with atopic dermatitis (AD) in the remission period.Methods A multicenter,randomized,parallel-group,controlled clinical trial was conducted from December 2017 to September 2018.A total of 297 children aged 2-12 years with moderate AD were enrolled from 5 hospitals in China,and randomly divided into the test group (148 cases) and control group (149 cases).In the acute stage,the two groups were both topically treated with mometasone furoate cream once a day on the skin lesions,and with an emollient containing Prinsepia utilis Royle oil extracts and other extracts twice a day throughout the whole body for 2-4 weeks.The children would be enrolled into the remission stage if their Investigator's Global Assessment (IGA) score was ≤ 1 at following visits.In the remission stage,the test group was only topically treated with the emollient twice a day throughout the whole body,while mometasone furoate cream and the emollient were both withdrawn in the control group.At weeks 4,8 and 12 in the remission stage,the recurrence of AD,eczema area and severity index (EASI),children's dermatology life quality index (CDQOL) and adverse events were evaluated.Statistical analysis was carried out with SAS 9.4 software by using t test for comparison of normally distributed continuous data between two groups,chi-square test for comparison of unordered categorical data,Kaplan-Meier method for analysis of survival rates,Cox regression analysis for evaluating the effect of different therapies on AD recurrence in children in the remission stage,and Logistic regression analysis for analysis of odds ratio (OR) of EASI or CDQOL at week 4 in the remission stage between the test group and control group.Results Of the 297 children with AD,31 breached the clinical trial protocol,and 266 were included in the per protocol set (PPS),including 132 in the test group and 134 in the control group.In the PPS,114 and 106 patients completed the follow-up in the test group and control group respectively,and the recurrence rate was significantly lower in the test group (47,41.23％) than in the control group (84,79.25％;x2 =32.96,P ＜ 0.001).The time to recurrence was significantly longer in the test group(61.99 d ± 2.80 d)than in the control group(39.17 d ± 2.54 d,t =6.03,P ＜ 0.001),and the recurrence risk was significantly lower in the test group than in the control group (Log rank test,x2 =32.02,P ＜ 0.001).After adjustment for age and gender,Cox regression analysis showed that the recurrence risk in the test group was 0.35 times that in the control group (HR =0.35,95％ CI:0.24-0.51,P ＜ 0.01).At week 4 in the remission stage,the EASI score at P50-P75 and P75-P100 in the test group were 0.42,0.25 times that in the control group respectively (95％ CI:0.20-0.86,0.12-0.54 respectively;P =0.02,＜ 0.01respectively).Moreover,the CDQOL score at P75-P100 in the test group was 0.33 times that in the control group (95％ CI:0.17-0.65,P ＜ 0.01).No significant difference in the incidence of adverse events was observed between the two groups (P ＞ 0.05).Conclusion Maintenance treatment with the emollient containing Prinsepia utilis Royle oil extracts and other extracts can markedly reduce the recurrence risk in AD children,improve clinical symptoms,and enhance the quality of life.