Objective To observe the adipocytic differentiation potential of bone marrow stromal cells (BMS), and the effect of simvastatin on adipocytic differentiation of bone marrow stromal cells in vitro, and to elucidate the mechanisms of anabolic effect of simvastatin on bone formation. Methods BMS from femur and tibia of adult female BALB C mice were cultured in vitro. Changes of alkaline phosphatase (ALP) activity were determined after treatment with adipogenetic agonist (hydrocortisone 0 5 ?mol/L and indomethacin 60 ?mol/L, HI) for 6 days. Thenexpression of lipoprotein lipase (LPL) mRNA was detected by RT PCR after treatment with HI and different concentration of simvastatin for 72 h. Adipogenetic differentiation were also observed with Oil Red O staining and fluorescence activated cell sorting (FACS) after treatment with HI and different concentration of simvastatin or 100 ?g/L rhBMP 2 for 12 days. Results After BMS were treated with HI for 6 days, ALP activity was significantly decreased ( P

Objective To investigate the protective effect of Qingkailing (QKL) in cardiac muscle's injury induced by endotoxin and discuss the mechanism from the inflammatory factors expression and free radical metabolism. Methods A total of 48 male ICR mice were divided into normal group, model group, low dose group and high dose group, 12 mice in each group. The QKL(9 ml/kg) was administered via gavage daily for 4 days in low dose group, the QKL (18 ml/kg) was administered via gavage daily for 4 days in high dose group, the equivalent volume of saline was administered via gavage daily for 4 days in normal group and model group. At the fifth day all groups except normal group, received intraperitoneal injection of LPS 0.2 ml (40 mg/kg), and the normal group received equivalent volume of saline. Intragastric administrated again 0.5h, 8h and 20 h after the model establishment. We took blood from hearts 1 hour after the last administration. The QR-PCR was used to detect the expression of inflammatory factors. The Elisa was used to detect IL-1β, IL-6, TNF-α, MDA, SOD and GSH-Px. Results Compared to the model group, the content of TNF-α (68.75 ± 7.73 pg/ml, 62.03 ± 16.09 pg/ml vs. 116.06 ± 21.06 pg/ml), IL-1β (110.84 ± 40.61 pg/ml, 105.51 ± 38.21 pg/ml vs. 167.53 ± 54.82 pg/ml) and IL-6 (68.78 ± 20.57 pg/ml, 59.71 ± 13.59 pg/ml vs. 108.80 ± 28.21 pg/ml) in low dose group and high dose group were significantly decreased (P<0.01). The expression of TNFα mRNA (1.42 ± 0.15, 1.30 ± 0.46 vs. 3.00 ± 0.82),IL-1β mRNA (1.20 ± 0.57, 1.01 ± 0.40 vs. 2.32 ± 1.39) and IL-6 mRNA (1.53 ± 1.10, 1.16 ± 1.09 vs. 4.12 ± 2.23) in low dose group and high dose group were significantly decreased (P<0.01 or P<0.05). The content of MDA (10.64 ± 2.91 nmol/mg, 11.36 ± 3.02 nmol/mg vs. 15.21 ± 2.31 nmol/mg) in low dose group and high dose group were significantly decreased (P<0.01). The content of SOD (282.32 ± 35.90 U/mg, 325.07 ± 34.76 U/mg vs. 249.01 ± 45.22 U/mg) and GSH-Px (48.26 ± 17.13 U/g, 49.66 ± 22.11 U/g vs. 26.47 ± 20.37 U/g) in low dose group and high dose group were significantly increased (P<0.01). Conclusions The QKL plays a protective role in myocardial injury induced by endotoxemia. Its mechanism may be associated with down-regulation of expression of inflammatory factors, reducing free radicals and improvement of antioxidation.

Epidermal growth factor receptor (EGFR) and its ligands (EGF and TGFalpha) are over-expressed in a variety of tumors. Immunization EGF-carrier protein inhibits tumor growth through abrogating binding of EGF to EGFR. Here, a chimeric protein of EGF and TGFalpha (E5T) was genetically fused to Staphylococcal enterotoxin A (SEA), a bacterial superantigenic protein which promotes humoral B cell response through enhancement of Ag-specific CD4 T cells activity. The resulted fusion proteins were expressed in Escherichia coli and purified though metal chelating affinity chromatography. Immunization of E5T-mSEA fusion protein in mice induced production of high titers antibodies, which recognize both EGF and TGFalpha. Anti- E5T-mSEA serum at dilution of 1:10 significantly inhibited growth of A431 cell lines but had little effect on 293T cell lines.
Amino Acid Sequence ; Animals ; Cancer Vaccines ; biosynthesis ; immunology ; Cell Line, Tumor ; Enterotoxins ; biosynthesis ; genetics ; Epidermal Growth Factor ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Humans ; Immunization ; Mice ; Mice, Inbred C57BL ; Molecular Sequence Data ; Random Allocation ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Transforming Growth Factor alpha ; biosynthesis ; genetics

Objective To investigate the clinical features and treatment principles of inflammatory myofibroblastic tumor of the urinary bladder (IMTUB).Methods From April 2013 to October 2016,6 cases of IMTUB patients were analyzed retrospectively.All cases were presented with gross hematuria.4 cases underwent ultrasonography,of which 3 cases showed solid mass in bladder,1 case showed inflammatory change.6 cases underwent CT examination,3 cases with bladder cancer,1 case with bladder sarcoma,1 case with malignant transformation of adenoma,1 case with rich blood supply.No lymph node metastasis.Bladder occupying lesions were considered in 2 cases of MRI examination.5 cases of cystoscopy showed bladder solid mass.In 6 cases involved,2 patients received partial cystectomy,2 patients underwent transurethral resection of bladder tumor,1 patient underwent radical resection of urachal carcinoma and the other one was treated with chemotherapy.Results Immunohistochemical staining was positive in ALK (100.0％) 、Vimentin(100.0％) 、CK(100.0％) 、SMA (83.3％) 、EMA(66.7％) and Ki-67 (5％-30％),negative in S-100 and Desmin.Final pathological diagnosis was IMTUB.So far,neither recurrence nor metastasis has been detected for 6 ～ 42 months in 5 cases and the other one lost to follow-up.Conclusions IMTUB is a kind of rare benign tumor of bladder.The golden standard of diagnosis is pathological diagnosis.Surgical resection is the first choice for treatment.Recurrence and metastasis are after the surgery treatment.All patients should be followed up closely.

Objective To detect the dynamic visual acuity ( DVA) before and after vestibular habituation of subjects in order to optimize the DVA assessment criteria .Methods The vestibular function examination system was applied to the detection of static and dynamic visual function in 16 healthy subjects .Results When the speed of left or right swinging was fast enough , DVA before and after vestibular habituation was different .Conclusion Subjects with vestibular habituation can reduce their sensitivity to the vestibular system , the changes in DVA are better than before habituation , and the vestib-ular function adaptability training may have effect on DVA .

Objective:To construct a simple, precise and personalized comprehensive nomogram for prediction the risk of multidrug-resistant tuberculosis (MDR-TB) and to evaluate its prediction value among individuals with previous tuberculosis history (PTBH).Methods:A matched case-control study (1∶2 ratios) was performed in 1 881 patients with PTBH treated in 12 designated tuberculosis hospitals in Hangzhou City between January 1, 2005 and December 31, 2019, and there were 1 719 patients in training set, and 162 in validation set. A multivariable Cox regression analysis was used to evaluate independent predictors for the incident of MDR-TB in individuals with PTBH. A comprehensive nomogram was developed based on the multivariable Cox model. The accuracy of the prediction was assessed using concordance index (C-index), calibration curve and area under the receiver operator characteristic (ROC) curve.Results:The nomogram constructed based on the multivariable Cox regression model incorporated 10 independent predictors of the risk of MDR-TB. A history of direct contact (grade 1, 0-100.0 points) ranked on the top of all risk factors, followed by duration of positive sputum culture (grade 2, 0-84.5 points), unfavorable treatment outcome (grade 3, 0-52.0 points), human immunodeficiency virus infection (grade 4, 0-48.5 points), retreated tuberculosis history (grade 5, 0-40.0 points), non-standardized treatment regimens of retreated tuberculosis (grade 6, 0-32.5 points), duration of pulmonary cavities (grade 7, 0-31.0 points), passive mode of tuberculosis case finding (grade 8, 0-25.0 points), age<60 years (grade 9, 0-17.5 points), and standard frequencies of chest X-ray examination (grade 10, 0-14.0 points). The C-indexes of this nomogram for the training and validation sets were 0.833 (95% confidence interval ( CI) 0.807-0.859) and 0.871 (95% CI 0.773-0.969), respectively, indicating that the nomogram had good fitting effect. The calibration curves for the risk of incident MDR-TB showed an optimal agreement between nomogram prediction and actual observation in the training and validation sets, respectively.The areas under ROC curve of the 1-year, 5-year, and 10-year MDR-TB risk probability of the training set were 0.904, 0.921, and 0.908, respectively, and those of the validation set were 0.954, 0.970, and 0.919, respectively. Conclusion:Through this nomogram model, clinicians could precisely predict the risk of incident MDR-TB among individuals with PTBH in the clinical practice.

Objective:To evaluate A modified Blumgart pancreaticojejunostomy in laparoscopic pancreaticoduodenectomy.Methods:The clinical data of 81 patients undergoing laparoscopic pancreaticoduodenectomy in Zhengzhou University Affiliated Cancer Hospital from Jan 2019 to Jan 2022 were retrospectively analyzed. Among them, 26 patients underwent modified Blumgart anastomosis and 55 underwent conventional Blumgart anastomosis.The data of demographics, liver function, pancreatic texture, operational result and complications were compared between the two groups.Results:The preoperative data (body mass index, preoperative albumin, prealbumin, transaminase, total bilirubin) between two groups were comparable ( P>0.05). There was no significant difference in pancreatic texture, pancreatic duct diameter and intraoperative blood loss between the two groups ( P>0.05). The modified group had shorter total operation time ( P<0.05), shorter pancreaticojejunostomy time ( P<0.05), shorter postoperative hospital stay ( P<0.05). The incidence of total pancreatic fistula and biochemical fistula in the modified group were lower than those in the conventional group ( P<0.05). There was no significant difference in the incidence of B/C grade pancreatic fistula and bile leakage, postoperative bleeding, infection and delayed gastric emptying between the two groups ( P>0.05). Conlusions:The modified Blumgart pancreaticojejunostomy during laparoscopic pancreaticoduodenectomy is safe, easy to do and time saving. While the incidence of postoperative pancreatic fistula with clinical significance　and other major complications were similar to traditional Blumgart procedure.

Objective To investigate the epidemiological characteristics of non-toxic goiter and non-toxic thyroid nodules in the regions with different iodine intakes and the factors influencing the occurrence, development and outcome of goiter and thyroid nodules. Methods 3 385 subjects, who had taken part in the previous survey in 1999 with the ultrasonic examination of thyroid, were composed of individuals in Panshan with chronic mild iodine deficiency,in Zhangwu with more than adequate iodine "after iodine supplementation and in Huanghua with excessive iodine. These 3 groups of subjects were followed up in 2004. Results (1) The cumulative incidences of diffuse goiter in Panshan ,Zhangwu and Huanghua were 7.1% ,4.4% and 6.9% ,respectively ,being the lowest in Zhangwu (P<0.01) and those of nodular goiter were 5.0% ,2. 4% and 0.8%, respectively, being the highest in Panshan (P<0.01). (2) The incidences of single nodule were 4.0% ,5.7% and 5.6%, respectively, and those of multiple nodules 0.4%, 1.2% and 1.0%, respectively. (3)The result of logistic analysis showed that iodine deficiency,iodine excess and positive thyroid autoantibodies (TAA) were the independent risk factors for the occurrence of goiter. (4)In Zhangwu ,the incidence of non-toxic goiter in the group with positive TAA was higher than that in the group with negative TAA(P<0.01) ,while there were no such differences in Panshan and Huanghua. (5)In these three regions, the rates of positive TAA in the individuals with diffuse non-toxic goiter were higher than those in the healthy subjects (P<0.05). And in Huanghua,the rate of positive TAA in subjects with non-toxic nodular goiter was also higher than that in the healthy individuals (P<0.05). Conclusion Iodine deficiency and iodine excess may both induce the raising incidence of goiter. Nodular goiter is prevalent in iodine deficient district and diffuse goiter is the predominant form in places with iodine excess. Thyroid autoimmunity is associated with occurrence and maintenance of goiter, and this phenomenon is more obvious in the community with previous iodine deficiency followed then by treatment with more than adequate iodine.

Objective:To investigate the role of nicotinamide (NIC) in the differentiation of neural crest cells from human embryonic stem cells (hESCs), and lay the foundation for the induction of hESC-derived corneal endothelial cells.Methods:hESCs line H1 cultured for 5-7 days was used for induction.According to the different components of the neural crest induction medium, cells were assigned into different groups for 7-days induction, including group treated without NIC cultured in induction medium only, group treated with NIC cultured in induction medium containing 10 mmol/L NIC, NIC+ resveratrol (Res) group cultured in induction medium containing 10 mmol/L NIC and 10 μmol/L Res and Sirtinol group cultured in induction medium containing 10 μmol/L Sirtinol.Res and Sirtinol were used as SIRT1 activity agonist and inhibitor, respectively.The relative mRNA expression levels of hESCs and neural crest cell markers were detected by real-time fluorescence quantitative PCR at 1, 3, 5 and 7 days during the induction.The expression of neural crest cells markers after 7 days of induction was assayed by immunofluorescence staining.The induction efficiency of NIC and the effect of SIRT1 regulation on human natural killer 1 (HNK-1) positive cells expression were evaluated through flow cytometry analysis of percentages of nerve growth factor receptor (P75) and HNK-1 + cells. Results:Compared with the group treated without NIC, the mRNA expressions of totipotent genes octamer transcription factor 4 (OCT4) and homeodomain proteins (NANOG) were significantly decreased, and the mRNA expression levels of neural crest cell markers P75, HNK-1, SRY-related HMG box (SOX) 9 and SOX10 were significantly increased in the group treated with NIC after 5 days of induction (all at P<0.05). In the group treated without NIC, P75 was weakly expressed, and HNK-1 was sporadically expressed, and transcription factor AP-2β (AP-2β) and paired-like homeodomain transcription factor 2 (PITX2) were not detected.In the group treated with NIC, P75, HNK-1, AP-2β and PITX2 were strongly expressed.The proportion of P75 + HNK-1 + cells and P75 + cells were both significantly higher in the group treated with NIC than without NIC ( t=8.481, P=0.001; t=2.987, P=0.041). The percentage of HNK-1 + cells in groups treated without and with NIC, NIC+ Res group and Sirtinol group were (34.267±12.522)%, (89.633±1.358)%, (64.667±6.429)% and (86.300±3.460)%, respectively, with a statistically significant overall difference ( F=36.799, P<0.001). The proportion of HNK-1 + cells in NIC+ Res group was significantly lower than that in the groups treated with NIC and Sirtinol (all at P<0.05). Conclusions:NIC promotes the differentiation of hESCs-derived neural crest cells by inhibiting the activity of SIRT1 to enhance the expression of HNK-1.NIC treatment may provide a new strategy for source of seed cells in the treatment of neural crest cell-related diseases, such as corneal endothelial transplantation.

Due to the serious shortage of corneal donor, the development of penetrating keratoplasty and corneal endothelial transplantation is severely restricted in clinical practice.The root cause is the limited proliferation capacity of healthy corneal endothelial cells.With the continuous development of tissue engineering technology and cell engineering technology, the research of tissue-engineered cornea has made some progress. In vitro culture of corneal endothelial cells with high density and healthy endothelial function for transplantation is a hot topic in current tissue engineering research.The keys of tissue-engineered corneal endothelial technology include seed cells, vector materials and the strategy of cell transplantation.At present, many research teams domestic and abroad have reported that the source of seed cells includes human corneal endothelial cells, stem cells, vascular endothelial cells and human amniotic epithelial cells.Vector materials include amniotic membrane, acellular corneal stroma, posterior elastic layer, anterior capsular membrane and various biomaterials.The cultured cells are transplanted by penetrating keratoplasty, corneal endothelial transplantation or anterior chamber injection.This review summarized the latest progress in the research on the source of corneal endothelial seed cells, the selection of vectors and the methods of corneal endothelial transplantation, and summed up the problems faced in the current research and looked forward to its prospects.