Over one half the patients with rheumatoid arthritis (RA) are being treated with methotrexate (MTX). Although well proven, the efficacy of MTX varies in individual patients. This study examined the metabolic biomarkers that can be used to predict the therapeutic effect of MTX by using metabolomic analysis. Rats were immunized with collagen to rapidly cause collagen-induced arthritis (CIA) and then treated with 0.1 mg/kg MTX for 4 weeks. The clinical signs and the histopathological features of CIA were observed to evaluate the therapeutic effects. Urine samples of CIA rats were collected, and analyzed by using 600 M (1)H-nuclear magnetic resonance ((1)H-NMR) for spectral binning after the therapy. The urine spectra were divided into spectral bins, and 20 endogenous metabolites were assigned by Chenomx Suite. Multivariate analyses were performed to identify the spectral pattern of endogenous metabolites related to MTX therapy. The results showed that the clustering of the spectra of the urine samples from the responsive rats (n=20) was different from that from the non-responsive rats (n=11). Multivariate analysis showed difference in metabolic profiles between the responsive and non-responsive rats by using partial least squares-discrimination analysis (PLS-DA) (R(2)=0.812, Q(2)=0.604). In targeted profiling, 13 endogenous metabolites (uric acid, taurine, histidine, methionine, glycine, etc.) were selected as putative biomarkers for predicting therapeutic response to MTX. It was suggested that (1)H-NMR-based metabolomic analysis can be used to predict the therapeutic effect of MTX, and several metabolites were found to be related to the therapeutic effects of MTX.

Background To maintain corneal transparency is important for good visual function.A new treatment concept and the selection of surgical techniques and timing of surgery are critical for stopping the infringement of cornea tissue after alkali burning and other chemical warfare agents.Objective This study was to investigative the ultrastructure and histopathological status following the femtosecond laser-assisted deep lamellar keratoplasty (DLK) for acute alkali burn of cornea.Methods Acute corneal alkali burn models were established in 12 New Zealand rabbits by putting the 6 mm filter paper with 1 mol/L NaOH at the central cornea for 30 seconds.The rabbits were randomly allocated to femtosecond laser-assisted DLK group and model control group according to the randomized number table method.Femtosecond laser-assisted DLK was performed to transplant the corneal grafts of domestic rabbits to the model rabbits 24 hours after burning.The rabbits were sacrificed 1 week,2 weeks and 4 weeks after modeling,and the corneas were extracted for the preparation of corneal section.The cornea were performed with hematoxylin and eosin staining to assess the histopathological status under the optical microscope,and the ultrastructure of grafts and corneas was examined under the transmission electron microscope (TEM).Results Acute corneal alkali bourn models were successfully eatablished.In the fourth week after surgery,corneal graft was clear in the femtosecond laser-assisted DLK group.However,corneal swelling,conjunctival congestion and neovascularization were found in the model control group.Histopathological examination revealed the defect of corneal epithelium,edema of stroma,loose arrangement of collagen fibers,much vacuoles,few neovascularization and infiltration of a large number of inflammatory cells in the model control group,but in the femtosecond laser-assisted DLK group,the inflammatory response was slight.More desmosomes among the endothelial cells were seen,and the nuclei were intact in the grafts.In the fourth week after surgery,the transplanted corneas were transparent with the regular arrangement of collagen fibers and entire fibroblasts in the femtosecond laser-assisted DLK group under the TEM.However,flat surface corneal epithelial cells and shedding of some epithelial cells were exhibited in the modelcontrol group.Conclusions Femtosecond laser-assisted DLK can effectively alleviate the inflammatory response,promote epithelial healing and enhance intercellular tight junction in the cornea with acute alkali burn.

Background Graft rejection is a primary cause of corneal transplantation failure,especially in high-risk keratoplasty.How to extend the survival time of graft is a problem to be solved.Objective This study was to investigate the influence of immune tolerance on high-risk rat keratoplasty induced by donor bone marrowderived immature dendritic cells (imDCs) transfected by chemokine receptor 7 (CCR7) recombinant adenovirus (Ad).Methods Bone marrow-derived imDCs were isolated and cultured from femur marrow of one male Wistar donor rat.The cells were transfected using recombinant Ad vector with rat CCR7 gene and resuspended in 500 μl PBS containing 1％ fetal bovine serum with the cells 1 × 107.High-risk corneal transplantaion models were established using monolateral corneal alkali-burn method in 60 SD rat recipients, and then allograft keratoplasty was performed with the 30 Wistar rats as donors.The models were randomized into the PBS group,imDCs group,imDCs with blank Ad vector group and imDCs with Ad-CCR7 group following the corresponding solution injection via caudal vein on preoperative day 7 and postoperative day 3 respectively.The survival time of graft was evaluated under the slit lamp microscope once per day.On the 14th day after operation, corneal sections were prepared from 6 eyes of each group for the pathological examination,and the relative expression levels of T helper cell 1 (Th1)-related factors,interleukin-2 (IL-2) mRNA and interferon-γ(IFN-γ) mRNA,as well as Th2-related factors, IL-4 mRNA and IL-10 mRNA, were detected by reverse transcription PCR (RT-PCR).The use and care of animals complied with the ARVO statement Results The mean survival time of grafts was (10.44±1.88) , (16.00±2.18) , (15.11±2.03) and(23.67±2.83) days in the PBS group,imDCs group,imDCs with blank Ad group and imDCs with Ad-CCR7 group, respectively, with a significant difference among the 4 groups (F =53.005, P =0.000).Compared with the PBS group, the mean survival time of grafts was considerably extended in the imDCs group,imDCs with blank Ad group and imDCs with Ad-CCR7 group (t=5.220,4.385,12.423 ,all at P=0.000) ,and a remarkble prolongation of graft survival duration was seen in the imDCs with Ad-CCR7 group in comparison with the imDCs group and the imDCs with blank Ad group (t =7.204,8.039,both at P=0.000).The relative expression levels of IFN-γ mRNA and IL-2 mRNA in the grafts were significantly lower,but the relative expression levels of IL-4 mRNA and IL-10 mRNA were significantly higher in the imDCs group,imDCs with blank Ad group and the imDCs with Ad-CCR7 group than those in the PBS group (all at P＜0.05).Compared with the imDCs group and the imDCs with blank Ad group, the relative expression levels of IFN-γ mRNA and IL-2 mRNA in the grafts were remarkably delined,but the relative expression levels of IL-4 mRNA and IL-10 mRNA were remarkably elevated in the imDCs with Ad-CCR7 group (all at P =0.000).Conclusions Application of imDCs transfected with CCR7 recombinant Ad via vena caudalis can prolong the survival time of grafts after keratoplasty of SD rats probably by affecting the balance of Th1/Th2 cytokines.

ObjectiveTo study the regulatory effect of Tripterygium wilfprdii polyglycoside (TWP) on the expression of EGFR and ErbB-2 induced arthritis rats.The effect of TWP on arthritis was also explored.MethodsAfter the model of CIA rats were established,the expression of EGFR and ErbB-2 in the synovium and articular cartilage were tested by immunohistochemical stain and real time PCR.ANOVA was used for statistical analysis.ResultsThe protein and mRNA expression of EGFR and ErbB-2 in the synovium (EGFR 0.268±0.059,ErbB-2 0.25±0.04,EGFR mRNA:14.2±0.55,ErbB-2 mRNA 23.46±3.64) and articular cartilage (EGFR 0.193±0.018,ErbB-2 0.217±0.033,EGFR mRNA:4.16±0.50,ErbB-2 mRNA 9.23±0.66) of the model group were significantly higher than those of the control group(P＜0.01).After being treated with TWP and MTX,the protein and mRNA expression of the EGFR and ErbB-2 decreased markedly (P＜0.01).Conclusion EGFR and ErbB-2 may play an important role in the pathogenesis of arthritis development.The molecular mechanism that TWP can treat synovitis and bone destruction of RA is related to the inhibition of EGFR and ErbB-2.

Objective:To analyze the composition differences of intestinal microbiota in patients with colon cancer and rectal cancer.Methods:The fecal samples of 72 patients in the Second Affiliated Hospital of Harbin Medical University from July 2018 to January 2019 were collected, and they were divided into colon cancer group and rectal cancer group, 36 cases in each group. DNA from fecal samples was extracted, and then high-throughput sequencing was performed on DNA. Bioinformatics was used to analyze the diversity and composition differences of intestinal microbiota between the two groups, and the potential cancer-promoting mechanisms of the differential flora were also discussed.Results:From high-throughput sequencing, 2 356 560 original sequences and 32 730 high-quality sequences were obtained from 72 samples. The average length of the sample sequence was mainly in the interval of 401-460 bp. And 1 409 operational taxonomic units (OTU) were acquired after OTU species taxonomy annotation of all the sequences. Alpha diversity analysis showed that Shannon index of the rectal cancer group and the colon cancer group was 2.61±0.56 and 2.43±0.67, respectively, and the difference was statistically significant ( t = 1.229, P = 0.223); Simpson index of the rectal cancer group and the colon cancer group was 0.17±0.09 and 0.21±0.16, respectively, and the difference was statistically significant ( t = 1.449, P = 0.151). Differences analysis of both groups and linear discriminant analysis (LDA) showed at the phylum level, Firmicutes were more abundant in the intestine of patients with rectal cancer (LDA = 4.67, P = 0.014), while Proteobacteria were more abundant in the gut of colon cancer patients (LDA = 4.49, P = 0.042). From the perspective of class level, the abundance of Gammaproteobacteria was higher in the intestine of patients with colon cancer (LDA = 4.50, P = 0.033), while the abundance of Erysipelotrichia was higher in the intestine of patients with rectal cancer (LDA = 3.50, P = 0.035). At the order level, the abundance of Erysipelotrichales was higher in the intestine of patients with rectal cancer (LDA = 3.50, P = 0.035); at the family level, the abundance of Porphyromonadaceae was higher in the intestine of patients with rectal cancer (LDA = 3.97, P = 0.033). Conclusion:The compositions of intestinal microbiota in patients with colon cancer and rectal cancer are significantly different, indicating that the different floras may contribute to the progression of colon cancer and rectal cancer.

Objective To examine the association between child-supporting from their folks and elder abuse in China so as to provide evidence for prevention and control of elder abuse.Methods Based on the third survey on the Status of Chinese women,organized by the All-China Women's Federation and the National Bureau of Statistics,7 159 residents aged 65 and older were included and general information on supporting child and elder abuse were gathered.Chi-square test and logistic regression were used to investigate the association between supporting child from elderly and elder abuse.Results The overall prevalence of elder abuse was 6.71％.Risks of elder people being abused by family numbers varied from different supporting child situations.After adjusting for potential confounding factors,ORs for the elderly appeared as follows:OR=1.99 (95％CI:1.56-2.54) for those who only providing support to male children;OR=2.07 (95％CI:1.51-3.79) for those only providing support to the female offspring and OR=2.32 (95％CI:1.72-3.13) for those who did not support their children regardless of their sex identity.Elderly who provided support to their children on both sexes were exposed to lower risk of being abused than those who only supporting their male offspring.There was no significant difference appearing on the risk of elder abuse between those elderly who only supporting the male (OR=1.00) or the female offspring (OR=1.04,95％ CI:0.63-1.71),among all the participants in our study.However,such associations were different in urban and rural areas.Conclusions High prevalence of abuse was seen in China.The pattem of supporting child was associated with risk of elder abuse.Elderly who showed poor support to their children were under higher risk of being abused by their family members.

Objective To explore the neuroprotective effect of asiatic acid(AA)on brain damage after experimental subarachnoid hemorrhage(SAH)in rats.Methods A total of 108 adult SD rats were divided into the sham1 group,the SAH+vehicle group and the SAH+AA group,with 36 rats in each group.The 42 rats were divided into the sham2 group,3,6,12,24,48 and 72 h after SAH groups,with 6 rats in each group.Except the sham group,SAH model was established by unilateral external carotid artery puncture method in other groups.After modeling,the SAH+AA group was given AA solution(30 mg/kg)by gavage.Neurobehavioral changes were assessed by foot fault test and modified Garcia score.Western blot assay was used to detect the protein level of glutathione peroxidase 4(GPX4)in brain tissue.ELISA was used to detect the concentrations of glutathione(GSH)and malondialdehyde(MDA).Fluoro Jade B(FJB)staining was used to detect the neuronal death.Results Compared with the sham1 group,the SAH+vehicle group showed a significant increase in the proportion of empty steps and a significant decrease in the modified Garcia score,a significant decrease in GPX4 protein levels,a significant increase in MDA concentration(P＜0.05),a decrease in GSH concentration(P＜0.01)and a significant increase in the number of dead neurons(P＜0.05).Compared with the SAH+vehicle group,a significant decrease in the proportion of empty steps,a significant increase in the modified Garcia score,a significant increase in GPX4 protein level,a significant decrease in MDA concentration,a significant increase in GSH concentration(P＜0.05)and a significant decrease in the number of dead neurons in the SAH+AA group(P＜0.05).Conclusion AA may reduce brain injury after SAH in rats by inhibiting lipid peroxidation.

Over one half the patients with rheumatoid arthritis (RA) are being treated with methotrexate (MTX). Although well proven, the efficacy of MTX varies in individual patients. This study examined the metabolic biomarkers that can be used to predict the therapeutic effect of MTX by using metabolomic analysis. Rats were immunized with collagen to rapidly cause collagen-induced arthritis (CIA) and then treated with 0.1 mg/kg MTX for 4 weeks. The clinical signs and the histopathological features of CIA were observed to evaluate the therapeutic effects. Urine samples of CIA rats were collected, and analyzed by using 600 M (1)H-nuclear magnetic resonance ((1)H-NMR) for spectral binning after the therapy. The urine spectra were divided into spectral bins, and 20 endogenous metabolites were assigned by Chenomx Suite. Multivariate analyses were performed to identify the spectral pattern of endogenous metabolites related to MTX therapy. The results showed that the clustering of the spectra of the urine samples from the responsive rats (n=20) was different from that from the non-responsive rats (n=11). Multivariate analysis showed difference in metabolic profiles between the responsive and non-responsive rats by using partial least squares-discrimination analysis (PLS-DA) (R(2)=0.812, Q(2)=0.604). In targeted profiling, 13 endogenous metabolites (uric acid, taurine, histidine, methionine, glycine, etc.) were selected as putative biomarkers for predicting therapeutic response to MTX. It was suggested that (1)H-NMR-based metabolomic analysis can be used to predict the therapeutic effect of MTX, and several metabolites were found to be related to the therapeutic effects of MTX.
Animals ; Antirheumatic Agents ; administration & dosage ; Arthritis ; chemically induced ; drug therapy ; urine ; Biomarkers ; urine ; Collagen Type II ; Dose-Response Relationship, Drug ; Immunosuppressive Agents ; administration & dosage ; Magnetic Resonance Spectroscopy ; methods ; Male ; Metabolome ; Methotrexate ; administration & dosage ; Proteome ; analysis ; Protons ; Rats ; Rats, Sprague-Dawley ; Reproducibility of Results ; Sensitivity and Specificity ; Treatment Outcome

Objectives:To investigate the clinical characteristics of noncontiguous spinal tuberculosis and the efficacy and prognosis of one-stage posterior debridement,bone graft fusion and internal fixation for the treat-ment of noncontiguous spinal tuberculosis.Methods:The clinical data of 31 patients with noncontiguous spinal tuberculosis treated in our hospital between July 2016 and May 2022 were retrospectively analyzed,in-cluding 18 males and 13 females,aged 49.5±27.5 years.There were 24 cases with 2 lesions and 7 cases with 3 lesions.Responsible vertebrae were clarified,and surgical lesions,fusion segments,and internal fixation methods were determined for each patient,so as to develop individualized surgical plans.The patients were followed up for 29.7±14.7 months(15-85 months).The operative time,intraoperative blood loss,and intraoper-ative and postoperative complications were recorded.Erythrocyte sedimentation rate(ESR)and C-reactive pro-tein(CRP)were examined and recorded before operation,at 1 month,3 months,and 1 year after operation,and at the last follow-up.Visual analogue scale(VAS)was used to evaluate the pain before operation,at 1 week,1 month,3 months,1 year after operation and at the last follow-up.Cobb angle was measured before operation,at 1 week after operation,and at the last follow-up.The American Spinal Injury Association(ASIA)classification was recorded before operation and at the last follow-up.Bridwell bone healing criteria were used to evaluate postoperative tuberculosis activity,symptom improvement,deformity correction,and bone healing at the last follow-up.Results:Among the 31 patients,20(65.4％)had only one lesion(65.4％),23(74.2％)were admitted to the hospital with pain as the main complaint,15(48.4％)had only pain symptoms during the course of the disease,11 cases(35.5％)had only one lesion with pain symptoms,and 18(58.1％)patients had at least one lesion missed at the initial diagnosis.All the patients were successfully operated.The operative time was 280.0±52.2min(165-330min),and blood loss was 567.7±332.0mL(150-1000mL).There were 4 cases of cerebrospinal fluid leakage and 3 cases of incision infection after operation,which were cured after symptomatic treatment.All foci of tuberculosis were cured without recurrence or retransmission.At pre-operation,1 month,3 months,1 year after surgery,and at the last follow-up,ESR was 41.5±26.3mm/h,16.3±13.4mm/h,12.5±6.3mm/h,11.4±5.2mm/h,and 9.2±3.1mm/h,and the levels of CRP were 32.8±23.2mg/L,7.3±5.6mg/L,6.2±4.1mg/L,5.1±3.7mg/L,2.8±2.3mg/L,which were both significantly lower after operation than those before operation(P＜0.05).The VAS score was 6.4±2.4,2.4±1.7,2.3±1.3,1.6±0.9,0.9±0.7,and 0.4±0.3 before operation,at 1 week,1 month,3 months,1 year after operation,and at the last follow-up,which was significantly improved after operation when compared with that before operation(P＜0.05).The Cobb angle was 25.7°±4.9° before operation,15.4°±2.1° at 1 week after operation,and 17.1°±2.3° at the last follow-up,and there were significant differences between the postoperative angles and preoperative angles(P＜0.05).Among the 10 patients with preoperative neurological impairment,1 patient with preoperative grade A recovered to grade C at the last follow-up.Among the 4 patients with preoperative grade B,1 patient recovered to grade C and 3 to grade D.Of the 5 patients with preoperative grade C,2 recovered to grade D and 3 to grade E.All 42 bone graft lesions achieved bone fusion at 6-12 months after operation.At the last follow-up,34 lesions healed in Bridwell grade Ⅰ and 8 in Bridwell grade Ⅱ.Conclusions:For patients with noncontiguous spinal tuberculosis,one-stage posterior debridement,bone graft fusion and internal fixation is safe and efficient after determining responsible vertebrae and lesion features,which can obtain satisfactory results.

Objective:To study the use of perfluorobutane contrast-enhanced ultrasound (CEUS) in preoperative detection of microvascular invasion (MVI), and postoperative short-term recurrence of hepatocellular carcinoma (HCC).Methods:Patients who underwent hepatectomy with curative intent at the Chinese PLA General Hospital from January 2021 to April 2021 were prospectively enrolled into this study. Of 42 patients in this study, there were 36 males and 6 females, with age of (56.51±11.95) years old. All patients underwent preoperative perfluorobutane CEUS, and the characteristics of ultrasound, the vascular phase and Kupffer phase of perfluorobutane CEUS were recorded. Based on the pathological results, these patients were divided into the MVI and non-MVI groups. These patients underwent liver MRI once every 3 months postoperatively to diagnose tumor recurrence. According to the recurrence of HCC 6 months after operation, these patients were divided into the non-recurrence and the recurrence groups. Independent risk factors for MVI and short-term recurrence were analyzed by univariate and multivariate analyses.Results:Two patients had two lesions, and the remaining 40 patients had a single lesion. The pathological diagnosis of all the lesions were HCC (14 patients in the MVI group and 28 patients in the non-MVI group). The median follow-up was 6 (3, 6) months, and there were 8 patients in the recurrence group and 34 patients in the non-recurrence group. On logistic analysis, independent risk factors for MVI included the number of vessels detected on color Doppler flow imaging (CDFI) ( OR=5.762, 95% CI: 1.597-20.785, P=0.007), increased tumor size by more than 10% after CEUS arterial enhancement ( OR=10.186, 95% CI: 3.647-28.447, P=0.037), and thickness of corona enhancement at Kupffer phase of greater than 5 mm ( OR=17.340, 95% CI: 6.124-49.095, P=0.040). Cox regression showed the independent risk factors for short-term recurrence to include the number of vessels in CDFI ( RR=7.519, 95% CI: 1.086-52.051, P=0.041) and thickness of corona enhancement at Kupffer phase of greater than 5 mm ( RR=10.623, 95% CI: 1.265-89.218, P=0.030). Conclusion:Preoperative perfluorobutane CEUS had potential values in detecting MVI and in predicting postoperative short-term recurrence of HCC.