OBJECTIVE To explore the therapeutic effect of basic fibroblast growth factor (bFGF) coacervates on diabetic peripheral neuropathy (DPN) in diabetic rats.METHODS Poly (ethylene argininylaspartate diglyceride) (PEAD), heparin and bFGF were dissolved in saline at the mass ratio of 50:10:1 to obtain bFGF coacervates. The loading efficiency of bFGF in the coacervates was analyzed by Western blotting. The release profile of bFGF from the coacevates was detected by ELISA. Male SD rats were ip injected with streptozotocin 65 mg · kg- 1 to establish a diabetic model,and DPN occurred 8 weeks later. The DPN rats were randomly divided into free-coacervate group, bFGF group and bFGFcoacervate group. For bFGF group, bFGF 200 μg·kg-1 was im injected once daily for 3 d. In bFGF-coacervate group, bFGF coacervate solution (244 μL) equal to bFGF 200 μg · k - 1, was im given only once. DPN rats in free- coacervate group were im given the same volume of vehicle(PEAD + heparin) only once. Ten normal age peer rats were taken as normal control group.Footprint analysis was conducted each week to evaluate motor function. On the 30th day after treatment,the rats were sacrificed, and sciatic nerves of both sides were harvested for pathological observation through HE staining. Apoptosis in nerve tissue was detected by DAPI staining, and Ki67 and proliferating cell nuclear antigen (PCNA) protein levels were detected by Western blotting. RESULTS Western blotting and ELISA analysis indicated that bFGF-coacervates were well prepared at a mass ratio of 50:10:1,and controlled bFGF release for at least 35 d. The result of rat behavior evaluation and pathological index test indicated that, compared with normal control group, the sciatic function index (SFI) in free-coacervate group decreased significantly(P<0.01), the internal nerve fibers were accompanied by irregularity and serious demyelination, and there was a large number of apoptotic nuclei and low expressions of Ki67 and PCNA proteins (P<0.01).After injection with bFGF or bFGF-coacervates, the SFI increased progressively (P<0.05, P<0.01), and the proportion of fibers with myelin abnormalities and apoptotic cells was significantly reversed. Moreover, the levels of Ki67 and PCNA was evidently enhanced on the 30th day post- operation (P<0.05, P<0.01). Compared with bFGF group, the results of those detection indicators in bFGF-coacervate group were better (P<0.05, P<0.01). CONCLUSION PEAD and heparin complex can load bFGF with high efficiency, and control its release in a steady manner. For DPN rats,treatment with bFGF-coacervates is more effective than bFGF alone.

The osteogenic in vitro effect of low intensity pulsed ultrasound (LIPUS) on SD rat adipose-derived stem cells (ADSCs) was investigated. Rat ADSCs underwent LIPUS (intensity=100 mW/cm(2)) or sham exposure for 8 min per treatment once everyday in vitro, and then the alkaline phosphatase (ALP) activity and mineralized nodule formation were assessed to evaluate the osteogenic effect of LIPUS on ADSCs. To further explore the underlying mechanism, the osteogenic-related gene mRNA expression was determined by using reverse transcriptase-polymerase chain reaction (RT-PCR) at 1st, 3rd, 5th, 7th day after exposure repectively. Westen blot was used to evaluate the protein expression levels of two osteogenic differentiation associated genes at 7th and 14th day repectively. It was found that ALP activity was increased after LIPUS exposure and LIPUS resulted in mineralized nodule formation of ADSCs in vitro. LIPUS-treated ADSCs displayed higher mRNA expression levels of runt-related transcription factor 2 (Runx2), osteocalcin (OCN), ALP and bone sialoprotein (BSP) genes than controls, and the protein levels of Runx2 and BSP were also increased. The results suggested that LIPUS may induce the osteogenic differentiation of ADSCs in vitro.

Objective To explore the influence of the killer cell immunoglobulin like receptor (KIR) gene polymorphism on cytomegalovirus (CMV) infection and pathogenesis after hematopoietic stem cell transplantation (HSCT).Methods The KIR genotype was determined by sequence-specific primer polymerase chain reaction (PCR-SSP) in 138 pairs of donors and recipients before HSCT during October,2005 and May,2011.Posttransplant monitoring for CMVpp65 antigen was performed by indirect immune histochemically assays since week 2 after transplantation.The differences between CMV positive group and negative group,inhibitive and active KIR of donors and recipients,and KIR haplotype frequency of donors and recipients were analyzed.Results There were no significant differences in frequency of KIR gene and haplotype AA,AB,BB between the donors and recipients.The frequencies of 2DS2 and 2DS4 * 003-007 of donors in CMV positive group were obviously lower than those in CMV negative group with significant differences(8％ vs 16％,P =0.0420;3％ vs 13％,P =0.0050).There was no significant difference in KIR gene between CMV positive group and CMV negative group.The CMV infection rates of haplotype AA,BB,AB donors were 64.38％,36.84％ and 50.00％,while CMV infection rates of haplotype AA,BB,AB recipients were 53.73％,46.15％ and 51.72％,respectively.The CMV infection rate was higher in the patients received KIR haplotype AA donor than in those received KIR haplotype BB donor (36.84％ vs 64.38％,P =0.0299).2DS4 * 003-007 and haplotype BB of donor were found associated with CMV infection in multifactor analysis.Conclusion KIR genotypes of donors are associated with CMV infection after HSCT.

Objective To investigate the efficiency and safety of allogeneic hematopoietic cell transplantation for malignant hematological diseases in patients older than 50 yeas of age. Methods From May 2002 to January 2010, 35 patients (＞ 50 years) with malignant hematological diseases received allogeneic hematopoietic cell transplantation. In 35 patients, 18 patients were conditioned with non-myeloablative regimen and 17 patients with myeloablative regimen. The outcome,engraftment and prognosis of allogeneic hematopoietic cell transplantation were analyzed. Results The hematopoetic reconstitution was achieved in 32 of 35 patients. The median time of granulocyte count exceeding 0. 5 × 109/L was 12 days and the that of platelet count exceeding 20 × 109/L was 17days. The cumulative incidence of aGVHD was 48. 6 %, and 37. 9 % patients developed cGVHD.The estimate probability of cumulative survival at 5 years was 48. 5 %, The estimate probability of cumulative mortality rate was 51.5 %, and the estimated transplant-related mortality was 22. 9 %.The relapse rate was 11.4 %. There was significant difference except for the incidence of cGVHD.Conclusion Allogeneic hematopoietic cell transplantation may be appropriate for older patients with malignant hematological diseases.

The osteogenic in vitro effect of low intensity pulsed ultrasound (LIPUS) on SD rat adipose-derived stem cells (ADSCs) was investigated.Rat ADSCs underwent LIPUS (intensity=100 mW/cm2) or sham exposure for 8 min per treatment once everyday in vitro,and then the alkaline phosphatase (ALP) activity and mineralized nodule formation were assessed to evaluate the osteogenic effect of LIPUS on ADSCs.To further explore the underlying mechanism,the osteogenic-related gene mRNA expression was determined by using reverse transcriptase-polymerase chain reaction (RT-PCR) at 1st,3rd,5th,7th day after exposure repectively.Westen blot was used to evaluate the protein expression levels of two osteogenic differentiation associated genes at 7th and 14th day repectively.It was found that ALP activity was increased after LIPUS exposure and LIPUS resulted in mineralized nodule formation of ADSCs in vitro.LIPUS-treated ADSCs displayed higher mRNA expression levels of runt-related transcription factor 2 (Runx2),osteocalcin (OCN),ALP and bone sialoprotein (BSP) genes than controls,and the protein levels of Runx2 and BSP were also increased.The results suggested that LIPUS may induce the osteogenic differentiation of ADSCs in vitro.

Objective:To analyze the expression of endoplasmic reticulum lipid raft-associated protein 2 (ERLIN2) in glioblastoma tissues, and to analyze its correlation with the clinicopathological features and prognosis of patients with glioblastoma.Methods:The TCGA database was used to analyze the expression level of ERLIN2 mRNA in glioblastoma. KM-plotter was used to analyze the correlation between ERLIN2 expression and prognosis in patients with glioblastoma. The expression level of ERLIN2 protein in 78 cases of glioblastoma tissue was determined by immunohistochemical staining, and then the tissue samples were divided into high expression and low expression groups according to the ERLIN2 expression level. The clinical and pathological data of patients were collected, including age, gender, tumor laterality, recurrence, isocitrate dehydrogenase(IDH) mutation, and the correlation between ERLIN2 protein and clinicopathological characteristics was further analyzed.Results:The mRNA of ERLIN2 was significantly overexpressed in glioblastoma, and the overall survival rate of patients with high ERLIN2 expression was low, suggesting a poor prognosis. The results of immunohistochemistry and clinicopathological analysis showed that ERLIN2 protein was significantly overexpressed in glioblastoma, and its expression level was significantly related to tumor recurrence.Conclusions:ERLIN2 protein is significantly overexpressed in glioblastoma tissues, and its expression level is related to the clinicopathological characteristics and poor prognosis, suggesting that ERLIN2 can be used as a potential prognostic factor for glioblastoma.

Objective:To investigate the predictive value of visceral fat area (VFA) on multiple metabolic risk factors in obese patients undergoing bariatric surgery.Methods:The retrospective case-control study was conducted. The clinical data of 146 obese patients undergoing bariatric surgery in the West China Hospital of Sichuan University from June 2015 to May 2020 were collected. There were 57 males and 89 females, aged (33±9)years, with a range from 15 to 65 years. All patients underwent multi-slice spiral computed tomography (CT) examination, and the total fat area (TFA), VFA and subcutaneous fat area (SFA) were measured. Observation indicators: (1) CT findings of abdominal fat area and patients with multiple metabolic risk factors; (2) receiver operator characteristic(ROC) curve of different clinical indicators for predicting multiple metabolic risk factors; (3) relationship between different VFA and clinicopathological features in patients; (4) analysis of factors influencing multiple metabolic risk factors in patients. Measurement data with normal distribution were expressed as Mean± SD, and comparison between groups was analyzed by independent sample t test. Measurement data with skewed distribution were expressed as M (range) and comparison between groups was analyzed by nonparametric rank sum test. Count data were represented as absolute numbers, and comparison between groups was analyzed using the chi-square test. The ROC curve was used to analyze the predictive value of different clinical indicators for multiple metabolic risk factors in patients. Logistic regression analysis was used for univariate and multivariate analysis. Results:(1) CT findings of abdominal fat area and patients with multiple metabolic risk factors. CT findings of 146 patients showed that VFA was (212±122)cm 2, SFA was (419±147)cm 2, visceral-to-subcutaneous fat ratio (VSR) was 0.60±0.54. The levels of systolic blood pressure (SBP), diastolic blood pressure, fasting blood glucose (FBG), triglyceride and highdensity lipoprotein cholesterol (HDL-C) were (131±16)mmHg(1 mmHg=0.133 kPa), (86±12)mmHg, (6.4±2.5)mmol / L, 4.43 mmol /L(range, 1.23-9.99 mmol/L), (1.5±1.3)mmol/L, respectively. Among the 146 patients, 85 had impaired FBG or diabetes, 82 had hypertension, 139 had high triglyceride, 91 had low serum HDL-C; 128 had multiple metabolic risk factors. (2) ROC curve analysis of different clinical indicators for predicting multiple metabolic risk factors: the ROC curve showed that VFA had better ability to predict or diagnose multiple metabolic risk factors in patients ( hazard ratio=0.617, 95% confidence interval as 0.470-0.764, P<0.05). The cutoff value of VFA was 163.52 cm 2 by calculating Yoden index in ROC curve. (3) Relationship between different VFA and clinicopathological features in patients: according to the cutoff value of 163.52 cm 2 in ROC curve, 146 patients were divided into high VFA group (≥163.52 cm 2) and the low VFA group (<163.52 cm 2), with 49 cases and 97 cases respectively. Cases with hypertension, level of SBP, cases with impaired FBG or diabetes, cases with multiple metabolic risk factors were 64, (134±17)mmHg, 63, 90 for the high VFA group, versus 18, (127±13)mmHg, 22, 38 for the low VFA group; there were significant differences in above indicators between the two groups ( χ2=11.309, t=6.916, χ2=5.380, 6.988, P<0.05). (4) Analysis of factors influencing multiple metabolic risk factors in patients: results of univariate analysis showed that VFA was a related factor for multiple metabolic risk factors in patients ( hazard ratio=3.722, 95% confidence interval as 1.341-10.328, P<0.05). Results of multivariate analysis showed that the VFA≥163.52 cm 2 was an independent rsik factcor or multiple metabolic risk factors in patients ( hazard ratio=5.182, 95% confidence interval as 1.441-18.641, P<0.05). Conclusions:VFA is positively correlated with hypertension, systolic blood pressure, impaired FBG and diabetes. VFA≥163.52 cm 2 is an independent predictor for multiple metabolic risk factors in obese patients undergoing bariatric surgery.

Objective: To explore the application and choice of latissimus dorsi musculocutaneous flap and thoracodorsal artery perforator flap in different wound repair. Methods: From March 2012 to February 2018, 8 cases of different wounds were repaired with island latissimus dorsi myocutaneous flap pedicled with dorsal thoracic artery, free latissimus dorsi myocutaneous flap, or thoracodorsal artery perforator flap combined with scapular flap. The patients includes 4 cases of trauma, 2 cases of tumor and 2 cases of osteomyelitis. Among them, 5 cases received pedicled grafting, 2 cases had anastomotic vascular free grafting combined with antibiotic bone cement chain bead, 1 case had thoracodorsal artery perforator flap combined with scapular flap. Results: All 9 flaps of 8 patients survived. The size of the flaps ranged from 22.0 cm×7.5 cm to 28.0 cm×21.0 cm. All the donor and recipient areas healed well. After 2 months to 2 years follow up, all flaps have good blood supply, and the limbs′ function was normal. The appearance of flaps were satisfactory, with fully treated osteomyelitis, and no recurrence of the tumor was observed. Conclusions According to wound characteristics, selective application of thoracodorsal artery perforator flap, pedicled or free latissimus dorsi myocutaneous flap is effective for the repair of muscle, skin and soft tissue defects, as well as osteomyelitis, after tumor resection.

ObjectiveTo investigate the effect and mechanism of Dahuang Zhechongwan （DHZCW） on adenine-induced renal fibrosis in rats from the perspective of intestinal flora. MethodThirty-six SD rats were randomly divided into a blank group， a model group， and high-， medium- and low-dose DHZCW groups （0.168， 0.084， 0.042 g·kg^-1）， and a pirfenidone group （200 mg·kg^-1）， with 6 rats in each group. Except for those in the blank group， rats in other groups were treated with adenine suspension （250 mg·kg^-1） by gavage for 28 days for renal fibrosis model induction. Subsequently， they received drug intervention for 4 weeks. Urine samples were collected from rats in metabolic cages， and renal function indicators including blood urea nitrogen （BUN）， urea， creatinine （Crea）， cystatin C （Cys C）， and 24-hour urine protein （24 h TP） were measured. Kidney samples were collected and subjected to hematoxylin-eosin （HE） staining and Masson's trichrome staining to observe the pathological changes in rat renal tissues. Western blot was used to detect the expression levels of key effector proteins α-smooth muscle actin （α-SMA）， type Ⅰ collagen （ColⅠ）， and type Ⅲ collagen （ColⅢ） in the kidneys. High-throughput sequencing of 16S rDNA was used to analyze the species diversity of rat intestinal flora. ResultCompared with the blank group， the model group showed increased BUN， urea， Crea， Cys C， and 24 h TP levels （P<0.01）. Compared with the model group， the high-， medium-， and low-dose DHZCW groups， as well as the pirfenidone group， showed significant reductions in BUN， urea， Crea， Cys C， and 24 h TP levels （P<0.01）， indicating that DHZCW intervention significantly improved renal function. In the model group， renal tissues exhibited significant fibrotic changes， and the protein levels of α-SMA， ColⅠ， and ColⅢ were significantly increased （P<0.01） compared to those in the blank group. Compared with the model group， the high-dose DHZCW group and the pirfenidone group had relatively normal tissue structure， with no significant pathological damage observed. However， fibrotic changes were observed in the medium- and low-dose DHZCW groups， with the changes being more significant in the low-dose group. The protein levels of α-SMA， ColⅠ， and ColⅢ were significantly decreased in the high-， medium-， and low-dose DHZCW groups， as well as the pirfenidone group （P<0.01）， indicating that DHZCW effectively reduced abnormal collagen deposition and inhibited renal fibrosis. From the perspective of intestinal flora， at the phylum level， compared with the blank group， the model group showed a significant increase in the abundance of Firmicutes and a decrease in Bacteroidetes， leading to a significant imbalance in their ratio. At the family level， the model group decreased the abundance of Lachnospiraceae， Prevotellaceae， and Bacteroidota_unclassified， and increased the abundance of Ruminococcaceae， Lactobacillaceae， and Oscillospiraceae. At the genus level， the model group showed significantly reduced abundance of Firmicutes_unclassified， Bacteroidota_unclassified， and Prevotellaceae_UCG-001， etc.， and increased abundance of UCG-005， Clostridia_UCG-014_unclassified， etc. Compared with the model group， DHZCW effectively reduced the abundance of potential pathogenic bacteria and increased the abundance of beneficial bacteria， regulating the intestinal flora. ConclusionDHZCW can effectively improve renal function and inhibit renal fibrosis， and its mechanism of action may be related to the regulation of intestinal flora.

ObjectiveTo explore the mechanism of Buzhong Yiqitang in ameliorating inflammatory injury in autoimmune thyroiditis (AIT) mice based on the Toll-like receptor 4（TLR4）/nuclear transcription factor-kappa B（NF-κB）/absent in melanoma 2（AIM2）inflammasome signaling pathway. MethodThe 120 genetically susceptible 8-week-old NOD.H-2^h4 mice were selected and randomly divided into control group， model group， low， medium and high dose groups of Buzhong Yiqitang （4.78， 9.56， 19.12 g·kg^-1）， and western medicine group （selenium yeast tablets， 3.033×10^-5 g·kg^-1）. The AIT model mice in each group drank ad libitum 0.05% sodium iodide aqueous solution for 8 weeks to establish the AIT model， and the control group drank ad libitum distilled water. Eight weeks later， the mice in each dosing group were divided into groups and gavage. The swelling of thyroid tissue was observed with the naked eye， and the weight of spleen was weighed. The content of serum inflammatory factors interleukin-1β （IL-1β）， interleukin-6 （IL-6）， tumor necrosis factor-α （TNF-α） was measured by enzyme-linked immunosorbent assay （ELISA）， and Real-time PCR was used to detect the expression of HMGB1， TLR4， AIM2， NF-κB p65，apoptosis-associated speck-like protein（ASC），cysteinyl aspartate-specific protease-1（Caspase-1）， IL-1β mRNA. Western blot was used to detect the expression of high motility group protein 1 （HMGB1）， TLR4， AIM2， NF-κB p65， phosphorylation（p）-NF-κB p65， ASC， Caspase-1， and IL-1β proteins in thyroid tissue， and immunofluorescence staining was used to observe the protein expression of HMGB1， AIM2， and NF-κB p65 in thyroid tissue of mice. ResultCompared with the control group， the thyroid tissue of mice in the model group was significantly swollen， the spleen quality was significantly increased， and the expression of HMGB1， TLR4， NF-κB p65， AIM2， ASC， Caspase-1， IL-1β in thyroid tissue was significantly increased （P<0.01）. Compared with the model group， the swelling of thyroid tissue in mice in each dose group of Buzhong Yiqitang was improved， the quality of spleen was significantly reduced， and the expression of HMGB1， TLR4， AIM2， NF-κB p65， p-NF-κB p65， ASC， Caspase-1， IL-1β in thyroid tissue was significantly reduced （P<0.05， P<0.01）. ConclusionBuzhong Yiqitang can effectively improve the inflammatory injury of AIT， and regulating the abnormal activation of the TLR4/NF-κB/AIM2 inflammasome signal pathway may be one of its intervention mechanisms.