BACKGROUND:There are various methodstoinduceadipogenic differentiation ofbone marrow mesenchymal stem cels, and the main componentfor adipogenic induction isindomethacin or rosiglitazone. However, there is a lack of comparative studyonthe induction efficiency and mechanism among these methods.
OBJECTIVE:Tocompare the adipogenic responses ofhuman bone marrow mesenchymal stem celsto different induction methods, and to analyze the mechanismunderlyingdifferent induction efficiency.
METHODS:After isolation and purification,the adipogenic abilitiesof human bone marrow mesenchymal stem cels in threedifferentculture systemswere comparedby oil red O staining and lipogenic geneassay. At 0, 1, 3 and 7 days of adipogenensis, mRNA expressionsof PPARγ, C/EBPα, Adiponectin and Leptin were detected.At7 daysofadipogenensis, protein expressionsof PPARγ and C/EBPβ were detectedby western blot assay,andeffects ofDIMIversusDIMRonphosphorylationofPPARγatSer273were compared.
RESULTS AND CONCLUSION:Findings from oil red O staining andreal-time PCRshowedthat DIMR significantlyinducedadipogenicdifferentiation of bonemarrow mesenchymal stem cels compared with DIM and DIMI at 7 daysofinduction. Western blot showed thattheprotein expressionsof PPARγ and C/EBPβ in the DIMIgroupwere significantly higher than those in the DIMRand DIM at 7days ofinduction. In addition, the ratio ofPPARγphosphorylation atSer273was lowerin the DIMR group thantheDIMI group.To conclude,DIMR has the most potential to induce early adipogenesis ofhumanbone marrow mesenchymal stem cels by weakening the phosphorylationof PPARγ-Ser273.

Objective To investigate the abnormal expression of proto-oncogene YES-associated protein (YAP) in gastric cancer tissues in the elderly and its correlation with poor prognosis.Methods Clinical data of 80 elderly patients with gastric cancer treated in our hospital from March 2011 to October 2014 were statistically analyzed.Results The positive expression rate of YAP was significantly higher in gastric carcinoma than in adjacent tissues [71.3％ (57/80) vs.13.8％ (11/80),P＜0.05].The positive expression of YAP were significantly associated gastric tumor size,tumor stage,invasion depth and lymph node metastasis (all P＜0.05),but had no correlation with tumor differentiation (P＞0.05).The 5-year survival rate was significantly lower in patients with YAP-positive expression than in patients with YAP-negative expression (P ＜ 0.05),but the differences in 1-year,3-year survival rates were not significant between the two groups (all P＞0.05)The YAP expression,tumor stage,lymph node metastasis were significantly associated with the prognosis of gastric cancer in patients (all P＜0.05).Conclusions YAP-positive expression rate is significantly higher in gastric cancer tissues than in adjacent tissues in the elderly,which indicates poor prognosis of patients with gastric cancer.

BACKGROUND:Recently, the effects of human umbilical cord mesenchymal stem cel s (hUCMSCs) and placenta-derived mesenchymal stem cel s (PDMSCs) on treatment of acute graft versus host disease (aGVHD) have been confirmed in some in vitro studies or animal models. But there are stil no reports comparing the therapeutic effects of these two cel types. OBJECTIVE:To compare the immunosuppressive function of hUCMSCs and PDMSCs in vitro or in a mouse aGVHD model. METHODS:(1) In vitro experiment. Human peripheral blood mononuclear cel s (PBMCs) were isolated and divided into four groups:PBMCs cultured alone, PBMCs stimulated with phytohaemagglutinin (PHA), PHA stimulated-PBMCs cocultured with hUCMSCs, PHA stimulated-PBMCs cocultured with PDMSCs. After 5 days, PBMCs proliferation and interferon-γlevel in cel supernatant were measured. (2) In vivo experiment. Fifty-seven BABL/C(H-2d) mice exposed to 8.5 Gy irradiation were randomly divided into five groups:only saline injection group, syngeneic bone marrow transplantation group, al ogeneic bone marrow transplantation group, aGVHD group, hUCMSCs treatment group, PDMSCs treatment group. The clinical aGVHD score, histopathology of skin, liver, and smal intestine, and survival time were analyzed at days 11, 14, 21 after transplantation. RESULTS AND CONCLUSION:(1) In vitro test:compared with the hUCMSCs, PDMSCs had stronger anti-inflammatory function. (2) In vivo test:The clinical scores on acute graft versus host disease were significantly lower in the hUCMSCs and PDMSCs treatment groups than that in the aGVHD group (P<0.05). The survival rates of mice were significantly increased in the hUCMSCs and PDMSCs treatment groups compared to the aGVHD group (P<0.05). Evident skin lesions were not found in al groups. Although smal intestine mucosal lesions were found in al groups, the damage level seemed similar. Notably, significant difference was found in the liver that multifocal necrosis and a large number of inflammatory cel s were seen in the aGVHD group, but less necrosis and inflammatory cel s in the hUCMSC and PDMSC treatment groups. In conclusion, hUCMSC and PDMSC are comparably effective in the treatment of aGVHD in mice.

Objective To investigate the effect of urinary training on urination control in infants using nappies after birth.Methods Stratified sampling method was used to investigate the daytime urination control and the using of diapers by children in kindergartens of 6 prefecture-level cities in Henan Province,China.The survey was conducted among parents of healthy children.Results In a total of 12 250 questionnaires,11 697 had response,and 10 562 valid questionnaires were collected,and the effective recovery rate was 86.22％.After the birth of infants,the age of toilet training was divided into 6 groups (0-＜ 3 months old group,3-＜ 6 months old group,6-＜ 12 months old group,12-＜ 18 months old group,18-＜ 24 months old group and no toilet training group).Results showed that toilet training within 12 months leads to higher urinary control rate compared with toilet training after 12 months and no toilet training till the age of 2 years old,70.56％ (4 831/6 847 cases) in contrast with 59.02％ (1 545/2 618 cases) and 42.48％ (466/1 097 cases),and the differences were statistically significant (x2 =114.76,335.48,all P ＜ 0.000 1).Within 12 months,there was no statistical difference in the urinary control rate between subgroups at 2 years of age(all P ＞0.05).After 12 months,the urination control rate decreased with the start time delayed till 2 years of age,and the urinary control rate at the age of 2 years old n different groups was 59.97％ (1 314/2 191 cases) and 54.10％ (231/427 cases) respectively.The results of other ages(0.5,1.0,1.5 years old) were similar to those of 2 years old.There was no difference between the groups of different genders (all P ＞ 0.05).Conclusions The use of diapers in infants before the age of 1 year to start urinary training is conducive to the development of urinary control in infants.

Objective To design a new integrated portable biliary internal-external drainage catheter carrying125 I seeds used for the treatment of malignant biliary obstruction lesions so as to achieve the dual curative effects of biliary drainage and brachytherapy. Methods A total of 15 patients with malignant obstructive jaundice, who were admitted to the First Affiliated Hospital of Zhengzhou University, China, during the period from September 2016 to January 2018, were enrolled in this study. Biliary stent implantation was performed in all patients, which was followed by insertion of a new integrated portable biliary internal-external drainage catheter carrying125 I seeds. The technical success rate, clinical success rate, complications, stent patency time and patient survival rate were evaluated. Results The placement of the drainage tube was simple and smooth, and the technical procedure was successful in all patients. One month after treatment, the bilirubin level was decreased significantly when compared with preoperative one (P<0.01), while the blood indexes and immunological indicators showed no obvious changes (P>0.05) . After treatment, 2 patients (13.3％) developed cholangitis and 2 patients (13.3％) had small amount of biliary bleeding, which returned to normal after symptomatic treatment. No severe complications such as perforation of bile duct, massive bleeding, radiation enteritis and radioactive source leakage, etc., occurred. The patients were followed up for 55-402 days, 6 patients (40.0％) developed biliary re-obstruction. The median patency time of stent was 255 days, and 6-month stent patency rate was 64.5％. Five patients died and 10 patients survived, the 9-month survival rate was 64.3％, the median survival time was 368 days. Conclusion By using the new integrated portable biliary internal-external drainage catheter carrying125 I seeds, the effects of bile drainage and brachytherapy can be simultaneously achieved. Preliminary clinical practice indicates that this new drainage catheter is feasible, safe and effective, although its long-term efficacy needs to be clarified with further follow-up observations and controlled studies.

Objective To study the effect of high copper on Wnt /β-catenin signaling pathway and oxidative stress. Methods BRL-3A cells were incubated with different concentrations of CuSO4.The cell growth and proliferation wre assessed using MRR method.The production of reactive oxygen species(ROS)and mitochondrial membrane potential (JC-1) were examined usingf flow cytometry. The content of malondialdehyde (MDA) and the activity of superoxide dismutase (SOD) were detected by microplate reader. The expression of protein was detected by Western Blot. Results①The results of MTT showed that CuSO4inhibited the growth and proliferation of BRL-3A cells in a time- and concentration-dependent manner(P<0.01).②Flow cytometry results showed that CuSO4induced a large amount of ROS and significantly decreased the fluorescence intensity of JC-1 in BRL-3A cells (P<0.01). ③ Microplate reader results showed that CuSO4increased the content of MDA and decreased the activity of SOD (P<0.05).④Western blotting assay showed that CuSO4significantly decreased the total expression levels of β-catenin and p-Ser 9-GSK-3β protein as well as nuclear levels of p-(S33+S37)-β-catenin and c-Myc (P<0.01) and increased expression levels of GSK-3β、DKK1、Dishevelled3 protein in BRL-3A cells. Conclusion High copper can induce oxidative stress and induce Wnt /β-catenin signaling pathway to deactivate liver cells,leading to hepatocellular injury.

Objective:To investigate the clinical characteristics and voice outcomes after laryngeal microsurgery for vocal fold epidermoid cysts coexisting with sulcus vocalis.Methods:The clinical data of 115 vocal fold epidermoid cysts coexisting with sulcus vocalis patients in Shandong provincial ENT hospital, were retrospectively analyzed, including 49 males and 66 females, aged 17-70 years old, and the duration of hoarseness ranged from 6 months to 30 years. All patients underwent surgery through suspension laryngoscope and microscope under general anestgesia. Ninety-four patients were treated with microflap excision of sulcus vocalis, cyst wall, and contents.And 21 patients that occulted with mucosal bridges were applied mucosal bridges resection (2 cases) and mucosal bridges reconstruction (19 cases) respectively. Videolaryngoscopy, subjective voice evaluation (GRBAS), objective voice evaluation, and Voice Handicap Index(VHI) were performed before and after surgery. All patients underwent histopathologic examination and follow-up after the procedure. The preoperative acoustic parameters of patients with vocal fold epidermoid cysts coexisting with sulcus vocalis were compared with those of vocal fold mucus retention cysts and simple vocal fold epidermoid cysts by independent samples t-test. The patients were compared by paired t-test for preoperative and postoperative parameters. Results:Significant reduction or lack of mucosal waves were shown via videolaryngostroboscopy in all 115 cases.In addition, vascular changes including dilation, tortuousness, increased branches, and abrupt direction change were shown on the cystic area. Eighty-one patients were detected cysts and/or sulcus vocalis by preoperative laryngoscopy, and intraoperative microscopic findings in the remaining 34 patients. The intraoperative microscopic examination revealed a focal pouch-like deficit plunging into the vocal ligament or muscle. The deep surface of the mucosal bridges was sulcus vocalis, and that in 89 cysts was lined with caseous content. Histopathology demonstrated a cystic cavity structure lined with squamous epithelium and caseous keratin desquamation inside the cystic cavity. Four of 115 patients were lost at follow-up and excluded from the analysis of voice outcomes after surgery. There was no significant mucosal wave and the voice quality in all but 14 patients 1month after surgery. Except for the fundamental frequency and noise harmonic ratio, all other voice parameters[ G, R, B, A, VHI-10, jitter, shimmer, maximum phonatory time (MPT) ]showed a significant improvement 3 months after surgery( t=15.82, 20.82, 17.61, 7.30, 38.88, 7.84, 5.88, -6.26, respectively, P<0.05). Then mucosal waves and the voice quality were gradually improved and became steady in 6 months after surgery. The subjective and objective voice parameters[G, R, B, A, VHI-10, jitter, shimmer, noise to harmonic ratio(NHR), MPT], except for the fundamental frequency, were all significantly improved( t=23.47, 25.79, 18.37, 9.84, 54.45, 10.68, 8.07, 3.24, -9.08, respectively, P<0.05). In addition, there were 2 patients with no significant improvement after the operation. Steady function with no complications was observed during the 12 months (up to 3 years in 34 patients) follow-up period in 111 patients. Conclusion:Ruptured vocal fold epidermoid cysts can result in sulcus vocalis and mucosal bridges. Characteristics changes in preoperative videolaryngoscopy are effective diagnostic tools. The complete excision of the cyst wall and repair of the lamina propria can lead to satisfactory long-term effects.

Purpose To investigate the clinical value of multi-center digital mammography radiomics nomogram model in predicting triple-negative breast cancer(TNBC).Materials and Methods The digital mammograms of 462 patients with pathologically confirmed breast cancer from November 2016 to March 2022 were retrospectively analyzed,including 243 cases from Yijishan Hospital of Wannan Medical College(institution 1),106 cases from Fuyang People's Hospital(institution 2)and 113 cases from Taihe People's Hospital(institution 3).According to the results of immunohistochemistry,a total of 349 breast cancer patients in institution 1 and institution 2 were randomly divided into the training group(244 cases,including 41 TNBC and 203 non-TNBC)and the validation group(105 cases,including 18 TNBC and 87 non-TNBC)according to the ratio of 7∶3,113 breast cancer patients(24 TNBC and 89 non-TNBC)from institution 3 were included in the external validation group.Comparing the mediolateral oblique and cranial cauda digital mammography images,the mammography imaging with larger lesion areas were selected,and the image segmentation and radiomics feature extraction were performed.The radiomics model was constructed by using Logistic regression.The clinicopathological parameters and radiomics scores were used to construct a nomogram.Receiver operating characteristic and decision curve analysis were used to evaluate the model performance.To compare The predictive performance between the models was compared.Results Finally,four radiomics features closely related to TNBC were selected to construct an radiomics model.The area under the curve,sensitivity and specificity of TNBC predicted by the radiomics model in training group,validation group and external test group were 0.868,90.24%and 72.91%,0.827,72.22%and 75.86%,0.837,70.83%and 78.65%,respectively.The area under the curve,sensitivity and specificity of TNBC predicted by the combined model in the training group,validation group and external test group were 0.903,80.49%and 86.70%,0.890,77.78%and 88.51%,0.870,62.50%and 85.39%,respectively.The combined model was better than the single image omics model in predicting TNBC,and the difference was statistically significant between the training group and the verification group(Z=2.061,2.064,both P<0.05),but not between the external test group(Z=1.223,P=0.221).In three group,decision curve analysis showed that the nomogram predicted a higher net benefit than the radiomics model for triple-negative breast cancer.Conclusion The radiomics model has high diagnostic efficiency in predicting TNBC,and the nomogram model combined with the radiomics score and histological grading can further improve the prediction efficiency.

OBJECTIVETo explore the formation of pre-metastatic niche in the mouse lung and to study the underlying molecular mechanisms whereby primary breast carcinoma-derived factors mediate recruitment of bone marrow-derived cells (BMDCs) and affect the formation of pre-metastatic lung environment before the arrival of tumor cells.

METHODSMammary carcinoma 4T1 cells were inoculated into the mammary gland to construct mouse model of breast cancer. Confocal microscopy was used to detect the recruitment of BMDCs in the pre-metastatic lungs. The expression of factors in the mouse sera and 4T1 cell culture media was assayed using RayBio Custom mouse cytokine antibody array kit. The mice were injected daily with recombinant VEGF for 7 consecutive days to observe the effect of VEGF on BMDCs recruitment in the mouse lung.

RESULTSNo BMDCs were observed in the lungs of control and 4T1-tumor-bearing mice on day 0. On day 7 and 14, clusters of BMDCs observed in the lungs of 4T1-tumor-bearing mice were 8.7±2.2/objective field and 48.8±3.2/objective field, respectively, significantly higher than those in the control mice (1.1±0.8/objective field and 3.1±1.7/objective field) (P<0.05 for both). Confocal microscopic observation found that metastatic breast cancer cells preferentially facilitate BMDCs recruitment sites in the pre-metastatic mouse lungs. The levels of VEGF, GM-CSF, and IL-6 in the serum of 4T1-tumor-bearing mice were significantly increased compared with those in the control group (P<0.05 for all). However, VEGF was detected only in the culture media of 4T1 cells. The amount of BMDCs in the mouse lung tissue was (22.8±3.6)/objective field in the VEGF group and (3.1±0.4)/objective field in the control group (P<0.05). There were 36.8±5.4 metastatic foci in the lung tissue of VEGF group and 12.6±2.2 in the control group (P<0.05).

CONCLUSIONSThe results of this study demonstrate that primary breast cancer cells can alter the lung microenvironment during the pre-metastatic phase and induce the formation of pre-metastatic niche. Primary tumor cell-derived VEGF may be a crucial factor responsible for the formation of pre-metastatic niche.

Animals ; Bone Marrow Cells ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Disease Models, Animal ; Female ; Granulocyte-Macrophage Colony-Stimulating Factor ; blood ; Humans ; Interleukin-6 ; blood ; Lung ; pathology ; Lung Neoplasms ; secondary ; Mice ; Recombinant Proteins ; administration & dosage ; Time Factors ; Tumor Microenvironment ; Vascular Endothelial Growth Factor A ; administration & dosage ; physiology ; secretion