ObjectiveTo investigate the effect of Liuwei Dihuangwan drug-containing serum （LDP） on epithelial-mesenchymal transition （EMT） and the expression of cancer stem cell （CSC） properties in 4T1 cells from triple-negative breast cancer by intervening myeloid-derived suppressor cells （MDSCs）. MethodsSPF-grade female SD rats were randomly divided into three groups， which were given 0.39， 1.94， 3.89 g·kg^-1·d^-1 suspension of Liuwei Dihuangwan for 7 days， respectively， to prepare low-， medium-， and high-dose LDPs. 4T1 cells were inoculated subcutaneously into the mammary glands of SPF-grade female Balb/c mice to construct a transplantation tumor model. Bone marrow cells were extracted from the tibia and femur and induced into MDSCs in vitro. The cell counting kit-8 （CCK-8） assay was used to detect the viability of 4T1 cells and MDSCs. The number of MDSCs and the expressions of CSC surface markers CD44 and CD24 in 4T1 cells were detected by flow cytometry （FC）. The migration， invasion， and proliferation of 4T1 cells were detected by cell scratch assay， Transwell invasion assay， and plate colony-forming assay， respectively. Western blot （WB） was used to detect the protein expression of transforming growth factor-β （TGF-β）， nuclear factor-κB （NF-κB）， C-X-C motif chemokine ligand 2 （CXCL2）， E-cadherin， and N-cadherin. The expression of EMT-related proteins E-cadherin and N-cadherin were detected by immunofluorescence （IF）. ResultsCompared with the normal group， LDP showed no significant inhibitory effect on the cell viability of 4T1 cells， but it significantly reduced the viability and number of MDSCs and reduced the number of MDSCs， as well as the expression of TGF-β （P<0.05， P<0.01）. The migration， invasion， and proliferation of 4T1 cells were increased after co-culture with MDSCs （P<0.05， P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin and the proportion of CSC （CD44⁺CD24^-） were elevated （P<0.05， P<0.01）， while the expression of E-cadherin was decreased （P<0.05）. After the intervention of MDSCs with LDP， followed by co-culture with 4T1 cells， the migration， invasion， and proliferation of 4T1 cells were obviously reduced （P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin were decreased （P<0.05， P<0.01）， and the expression of E-cadherin was increased （P<0.05， P<0.01）. There was no statistical difference in the proportion of CSC （CD44⁺CD24^-） in 4T1 cells. However， the proportion of CSC （CD44⁺CD24^-） was decreased in the co-culture system of 4T1 cells and MDSCs with LDP intervention （P<0.05， P<0.01）. ConclusionLDP can reduce the viability and number of MDSCs and the expression of TGF-β， NF-κB， and CXCL2， reverse EMT， and reduce the characteristic expression of CSC to inhibit the migration， invasion， and proliferation of 4T1 cells.

ObjectiveTo investigate the effect of Liuwei Dihuangwan drug-containing serum （LDP） on epithelial-mesenchymal transition （EMT） and the expression of cancer stem cell （CSC） properties in 4T1 cells from triple-negative breast cancer by intervening myeloid-derived suppressor cells （MDSCs）. MethodsSPF-grade female SD rats were randomly divided into three groups， which were given 0.39， 1.94， 3.89 g·kg^-1·d^-1 suspension of Liuwei Dihuangwan for 7 days， respectively， to prepare low-， medium-， and high-dose LDPs. 4T1 cells were inoculated subcutaneously into the mammary glands of SPF-grade female Balb/c mice to construct a transplantation tumor model. Bone marrow cells were extracted from the tibia and femur and induced into MDSCs in vitro. The cell counting kit-8 （CCK-8） assay was used to detect the viability of 4T1 cells and MDSCs. The number of MDSCs and the expressions of CSC surface markers CD44 and CD24 in 4T1 cells were detected by flow cytometry （FC）. The migration， invasion， and proliferation of 4T1 cells were detected by cell scratch assay， Transwell invasion assay， and plate colony-forming assay， respectively. Western blot （WB） was used to detect the protein expression of transforming growth factor-β （TGF-β）， nuclear factor-κB （NF-κB）， C-X-C motif chemokine ligand 2 （CXCL2）， E-cadherin， and N-cadherin. The expression of EMT-related proteins E-cadherin and N-cadherin were detected by immunofluorescence （IF）. ResultsCompared with the normal group， LDP showed no significant inhibitory effect on the cell viability of 4T1 cells， but it significantly reduced the viability and number of MDSCs and reduced the number of MDSCs， as well as the expression of TGF-β （P<0.05， P<0.01）. The migration， invasion， and proliferation of 4T1 cells were increased after co-culture with MDSCs （P<0.05， P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin and the proportion of CSC （CD44⁺CD24^-） were elevated （P<0.05， P<0.01）， while the expression of E-cadherin was decreased （P<0.05）. After the intervention of MDSCs with LDP， followed by co-culture with 4T1 cells， the migration， invasion， and proliferation of 4T1 cells were obviously reduced （P<0.01）. The expressions of NF-κB， CXCL2， and N-cadherin were decreased （P<0.05， P<0.01）， and the expression of E-cadherin was increased （P<0.05， P<0.01）. There was no statistical difference in the proportion of CSC （CD44⁺CD24^-） in 4T1 cells. However， the proportion of CSC （CD44⁺CD24^-） was decreased in the co-culture system of 4T1 cells and MDSCs with LDP intervention （P<0.05， P<0.01）. ConclusionLDP can reduce the viability and number of MDSCs and the expression of TGF-β， NF-κB， and CXCL2， reverse EMT， and reduce the characteristic expression of CSC to inhibit the migration， invasion， and proliferation of 4T1 cells.

Radiation enteritis is a kind of intestinal radiation injury in patients with pelvic and retroperitoneal malignancies after radiotherapy, and its occurrence and development process are very complicated. At present, studies have confirmed that intestinal microecological imbalance is an important factor in the formation of this disease. Abdominal radiation causes changes in the composition of the flora and a decrease in its diversity, which is mainly manifested by a decrease in beneficial bacterial species such as Lactobacilli and Bifidobacteria. Intestinal dysbacteriosis aggravates radiation enteritis, weakens the function of the intestinal epithelial barrier, and promotes the expression of inflammatory factors, thereby aggravating the occurrence of enteritis. Given the role of the microbiome in radiation enteritis, we suggest that the gut microbiota may be a potential biomarker for the disease. Treatment methods such as probiotics, antibiotics, and fecal microbiota transplantation are ways to correct the microbiota and may be an effective way to prevent and treat radiation enteritis. Based on a review of the relevant literature, this paper reviews the mechanism and treatment of intestinal microbes in radiation enteritis.

Objective: Ischemia-reperfusion (IR) injury is a leading cause of flap compromise and organ dysfunction during free-tissue transfer, and remains a great challenge for plastic surgeons. Thioredoxin-1 (Trx-1) was proved to protect the IR flap by mitigating the oxidative stress, and inhibiting the activation of apoptosis signal-regulating kinase-1 (ASK-1) and mitogen-activated protein kinase (MAPK) pathway. The aim of this study is to investigate the distinction of Trx-1 expression, apoptosis indices in different layers of IR flaps, and the feasibility of tissue-layer-specific administration of Trx-1. Methods: Ten patients′ specimens of IR flaps for DIEP breast reconstruction were collected and assessed for apoptosis and Trx-1 expression. Twenty mice were used to establish the IR flap model. The mice were sacrificed twenty-four hours after reperfusion. The flap tissues were harvested and tested by immunohistochemistry staining and TUNEL assay. The tissue-layer-specific dermoprotective effect of Trx-1 and the molecular mechanisms were assessed by an in vitro epithelial skin cell hypoxia-reoxygenation model. The statistics were conducted by t test and ANOVA using SPSS 20.0. Results: Trx-1 expression and apoptotic cells were observed mainly located in the basal layer of epidermis and the papillary layer of dermis in human IR flaps and mice models. Trx-1 depletion was 24.19 %± 2.23% in the basal layer of epidermis and the papillary layer of dermis of patient IR flaps, decreasing significantly compared with 70.71% ± 6.38% in control group (t = 27.54, P< 0.001). Similar tissue-layer-specific down regulation of Trx-1 also displayed in mice IR flap models (19.83% ± 2.34% vs. 76.59% ± 4.88%; t = 34.71, P<0.001). The apoptotic index in human samples significantly increased from 1.32% ± 1.52% in control group to 43.71 %± 3.17% in IR group (t =38.23, P<0.001); while it was proved to be dramatically raised in mice models from 0.86% ± 1.15% in control group to 41.14 %± 4.21% in IR group (t= 36.96, P < 0.001). Western Blot analysis revealed Trx-1 down regulation and a significant increase in ASK-1, p-p38, and c-PARP abundance in the hypoxia-reoxygenation-treated HaCaT cells (P < 0.01). Supplementation of recombinant human Trx-1 significantly reduced the apoptosis-related protein expression. Conclusions The basal layer of epidermis and the papillary layer of dermis are the main damaged tissue layers in the early stage of skin flap ischemia-reperfusion injury. The IR flap can be protected by precisely replenishing the vulnerable layers with Trx-1.

The expression levels of serum tenascin-C, osteopontin(OPN), and transforming growth factor-β1(TGF-β1) in the patients of diabetic mellitus were measured by ELISA. With the increase of the UACR, the expression of tenascin-C, osteopontin, and transforming growth factor-β1 showed a trend of increase and hypertension will argument this phenomenon. Pearson correlation analysis showed that levels of tenascin-C were positively correlated with HbA1C , body mass index, systolic blood pressure, diastolic blood pressure, UACR, osteopontin, and transforming growth factor-β1.

Objective To investigate the association of adaptor protein,phosphotyrosine interacting with PH domain and leucine zipper 1(APPL1)with urinary albumin excretion rate in patients with type 2 diabetes mellitus (T2DM),and to explore the role of APPL1 in the development of diabetic kidney disease(DKD). Methods According to the urinary albumin/creatinine ratio(UACR),288 newly-diagnozed patients with T2DM were divided into normal albuminuria group(UACR<30 mg/g,n=116),microalbuminuria group(UACR 30 ~300 mg/g,n=95),and macroalbuminuria group(UACR>300 mg/g,n=77). 130 healthy subjects with matched sex and age were used as control group. Serum APPL1,tumor necrosis factor α(TNF-α),and adiponectin levels were measured by ELISA method. Results Serum APPL1 level in T2DM patients was significantly higher than that in control subjects (P<0.01), and increased with the rising of UACR. In patients with T2DM, serum APPL1 level was negatively correlated with estimated glomerular filtration rate(r=-0.246, P<0.01) while it was positively correlated with HbA1C, low density lipoprotein cholesterol, total cholesterol, triglycerides, insulin resistance index, serum creatinine,blood urea nitrogen, systolic blood pressure, TNF-α, and adiponectin(r=0. 119, 0. 167, 0. 209, 0.194,0.273,0.242,0.131,0.144,0.365, and 0.952, respectively, P<0.05 or P<0.01). Conclusion Serum APPL1 level in patients with T2DM was increased with the rising of UACR, suggesting that APPL1 may be involved in the development of DKD.

267 newly-diagnosed patients with type 2 diabetes were divided into normoalbuminuria group [group N-UAlb, urinary albumin to creatinine ratio (UACR)<30 mg/g, n=103], microalbuminuria group(group M-UAlb, UACR 30~300 mg/g, n=88 ) , and macroalbuminuria group ( group L-UAlb, UACR>300 mg/g, n=76). The control group(group NC) consisted of 114 healthy individuals. Serum betatrophin, adiponectin(APN), and interleukin-1β( IL-1β) levels were determined with ELISA methods and the parameters of body mass index (BMI), estimated glomerular filtration rate(eGFR), HbA1C, fasting plasma glucose(FPG), OGTT 2h plasma glucose(2hPG), fasting insulin(FINS), OGTT 2h postprandial insulin(2hPINS), fasting C-peptide(FCP), homeostasis model assessment insulin resistant index(HOMA-IR), and blood lipid were collected. Compared with group NC, the serum betatrophin levels in patients with type 2 diabetes were obviously increased. In patients with type 2 diabetes, betatrophin levels increased along with the increase of UACR and there were significant differences in betatrophin among the three groups(P<0. 01). Betatrophin positively correlated with UACR, HbA1C, FPG, 2hPG, FINS, 2hPINS, HOMA-IR, TC, LDL-C, and TG( r were 0. 785, 0. 225, 0. 136, 0. 241, 0. 386, 0. 223, 0. 411, 0.216,0.193,and0.298,allP<0.05),and betatrophin were also positively correlated with APN and IL-1β(rwere 0. 643 and 0. 710, both P<0. 01). Stepwise multiple regression analysis showed that UACR, HbA1C, FINS, and TG were independent relevant factors affecting betatrophin levels.

Objective To investigate the expression levels of serum vasohibin-1(VASH-1)in type 2 diabetes mellitus(T2DM)patients at different stages of urinary albumin to creatinineratio(UACR)and to attempt to investigate the relationship between VASH-1 and inflammation and fibrosis in the pathogenesis of diabetic nephropathy(DN), one of the microvascular complications of T2DM. Methods 486 patients with T2DMwere divided into four groups:normal albuminuria [ UACR<30 mg/ g, n = 134], microalbuminuria [ UACR at 30-300 mg/ g, n = 122], clinical albuminuria [UACR > 300 mg/ g, n = 106 ], and clinical albuminuria hypertensive [ UACR > 300 mg/ g, with hypertension, n=124] groups. Age, course, serum levels ofVASH1, inflammation markers(CRP, ESR)and fibrosis marker( TGF-β1) with other biochemical indicators were measured, and 130 normal control subjects were also included. Results Compared with normal control group, the levels of UACR, HbA1C ,ESR, CRP, TGF-β1 and VASH-1 in groups ofnormal albuminuria, microalbuminuria, clinical albuminuria, and clinical albuminuria hypertensive were significantly higher(P<0. 05). Pearson correlation analysis showed that levels of VASH-1 were positively correlated with UACR, HbA1C ,ESR, CRPand TGF-β1( r = 0. 521, 0. 261, 0. 519, 0. 523, 0. 479, P<0. 001), while multivariate regression analysis showed that levels of UACR, HbA1C ,ESR, CRP and TGF-β1 were important factors affecting serum VASH-1 levels. Conclusion Serum levels of VASH-1 may become new biomarkers of early diagnosis of DN. Consequently, VASH-1 level may provide a new pattern and direction of inflammation and fibrosis for consideration in diabetic kidney damage.

OBJECTIVETo analyze the efficacy of percutaneous endoscopic gastrostomy (PEG) in pediatric patients.

METHODFrom October 2011 to October 2014, children in the gastrointestinal ward of Guangzhou Women and Children's Medical Center received PEG or jejunal tube PEG(JET-PEG). The success rate, operation time were recorded. The changes of their weight, enteral nutrition calories and the incidence of pneumonia before and after the first 6 months of operation were compared. Follow-up was conducted until October 2014, the recent and long term complications, the length of indwelling time, the replacement or removal of the tube were recorded, the patients swallowing function or the primary disease's outcomes were observed.

RESULTOf the 13 cases, 10 were male, 3 were female, their average age was 2 years (range 1.8 months-9 years). We performed PEG for 12 of the patients who had congenital craniofacial problems that led to feeding difficulties or recurrent cough and pneumonia (6/12), or neurological disorders (6/12) with inability to swallow, and in one case JET-PEG was performed, this child suffered from chronic intestinal pseudo-obstruction with vomiting and abdominal distension. The gastrostomy was successful in all the patients through one operation, the average operation time of PEG was (25 ± 3) minutes, JET-PEG was 60 minutes. One local skin infection was noted, no long-term complication occurred. In the first 6 months after operation, all the patients gained weight((5.5-30.5) kg postoperation vs. (3.0-30.0) kg preoperation), and 12 cases' enteral nutrition calories increased (from (209-502) to(272-543) kJ/(kg·d)), the incidence of pneumonia decreased in the children who had recurrent pneumonia before the operation (from (0-1.5) to (0-0.16) per month). Until October 2014, their average length of gastric tube indwelling time was 17.8 months (range 4-36 months). In 4 cases PEG tube was removed when they could eat completely independently, the other 9 needed enteral vein nutrition via PEG tube or jejunal tube, in 3 of them balloon type gastric fistula tube was applied. Two of the 13 cases who had cleft palate received stomatological operations when their weight grew to meet the standard.

CONCLUSIONPEG and JET-PEG are safe and effective method for enteric nutrition feeding in pediatrics, the technique causes minimal trauma and has rapid postoperative recovery, few complications, good aesthetic appearances and simple nursing, it can significantly improve their nutritional status and quality of life.

Child ; Child, Preschool ; Enteral Nutrition ; methods ; Female ; Gastrostomy ; adverse effects ; Humans ; Incidence ; Infant ; Male ; Nervous System Diseases ; therapy ; Pneumonia ; therapy

Objective To explore the associations of serum Mir-217 with silent information regulator 1 (Sirt1)and hypoxia-inducible factor-1α(HIF-1α)in type 2 diabetic patients with different urinary albumin excretion rates. Methods A total of 479 patients with type 2 diabetes were divided into normoalbuminuric(D1, n=181), microalbuminuric(D2, n=165), and macroalbuminuric(D3, n=133)subgroups. 192 normal subjects served as control group. Serum level of Mir-217 was detected by realtime PCR. Serum Sirt1, HIF-1α, and vascular endothelial growth factor ( VEGF ) levels were determined by enzyme-linked immunosorbent assay. Results Compared with control subjects, serum Mir-217 level was significantly increased in type 2 diabetic patients and gradually increased in D1, D2, and D3 groups(P<0. 01). The parameters of age, diabetes duration, body mass index, fasting plasma glucose, fasting insulin, homeostasis model assessment of insulin resistant index(HOMA-IR), HbA1C, low density lipoprotein-cholesterol( LDL-C) , total cholesterol ( TC ) , triglyceride ( TG ) , serum uric acid ( SUA ) , blood urea nitrogen(BUN), HIF-1α, VEGF, and Mir-217 all were positively correlated with ACR(all P<0. 05). High density lipoprotein-cholesterol(HDL-C)and Sirt1 were negatively correlated with ACR(both P<0. 05). VEGF, HIF-1α, Mir-217, BUN, diabetes duration, LDL-C, Sirt1, and SUA were independent factors that influenced ACR(all P<0. 01). Additionally, diabetes duration, HOMA-IR, HbA1C, ACR, LDL-C, TC, TG, SUA, BUN, HIF-1α, and VEGF were positively correlated with Mir-217(all P<0. 05), while Sirt1 was negatively correlated with Mir-217(P<0. 01). Conclusion Serum Mir-217, as a possible biomarker for early diagnosis of diabetic nephropathy, may be involved in the development of diabetic nephropathy by promoting chronic inflammatory reaction, renal fibrosis, and angiogenesis.