Objective To extract the Ginseng polysaccharide (GPS), polysaccharides of Tricholoma matsutake (PTM)and polysaccharide of Lentinus edodes (PLE)from gingeng, tricholoma matsutake and lentinus edodes respectively,and to analyze and identify their structures,and to prepare their complex,and to study the indirect antitumor activity invitro of polysaccharide complex.Methods The polysaccharides were extracted with hot water and precipitated by ethanol.The carbohydrate levels were determined by the method of phenol-sulfuric acid.The m-hydroxyphenyl method was used to determine the levels of uronic acid, and the national standard method was used to determine the levels of starch.Infrared spectroscope and chemical methods were performed to analyze their structures. Orthogonal experiment was used to study mixing methods. Cytotoxic T lymphocyte experiment and LDH release assay were performed to detect the influence of polysaccharide complex of GPS,PTM,and PLE in the CTL killing activity,and its indirect killing effect on the P815 cells.Results The extraction rates of GPS,PTM, and PLE were 8.85%,9.40%,and 10.50%;the levels of total polysaccharides were 62.96%,59.13%,and 33.86%;the levels of uronic acid were 16.44%,9.37%,and 16.44%;the starch levels were 7.26%,2.80%,and 3.77%,respectively.The identification results showed that the polysaccharides were obstrained.When the quality ratio of the three kinds of polysaccharides was 1∶1∶1 and the concentration was 600 mg·L-1 ,the CTL cytotoxicity was the highest.Conclusion The polysaccharide complex is obtained,identified and characterized. Polysaccharide complex can enhance the cytotoxicity of CTL and has the indirectly inhibitory effect on the proliferation of P815 cells.

Objective To explore mutation diagnosis and discuss the pathogenic and clinical characteristics of neurofibromatosis type 1 (NF1).Methods DNA sequencing combined with denaturing highperformance liquid chromatography (DHPLC) method was used to diagnose patients and parents.Results A new nonsense mutations c.503C ＞ G(p.S168 *) was identified.Conclusions NF1 is a rare autosomal dominant genetic disease.Most of them are caused by new mutations.Genetic diagnosis of sporadic cases is very important for treatment and the future generations.

Objective To explore the clinical characteristics of neonatal congenital cystic lung lesions. Methods Between January 2008 and June 2014,total 28 cases diagnosed congenital cystic lung lesions in neonatal center of Guangzhou Women and Children's Hospital were collected.The clinical data were analyzed including manifestations,lesion characteristics,imaging,diagnosis,treatment and prognosis,and the related literature were reviewed.Results There were 20 male and 8 female,16 cases of lobar emphysema,7 cases of pulmonary seques-tration,4 cases of congenital cystic adenomatoid malformation and 1 case of bronchogenic cyst.Main symptoms were dyspnea(78.57%),cyanosis (39.29%),wheezing cough (17.86%),feeding difficulties (14.29%),fever (10.71%),asymptomatic(21.43%).Two cases combined with congenital heart disease,4 cases combined with other malformations(such as diaphragmatic hernia,laryngeal stridor,funnel chest,polycystic kidney).There were lesions in chest CT image,4 cases underwent lobectomy,other cases underwent conservative treatment.Seventeen cases got better and discharge,3 cases dead.Conclusion The most common type of congenital cystic lung lesions in neonate is lobar emphysema.Main manifestations are dyspnea,cyanosis.If we suspect congenital cyst-ic lung lesions,we should do chest CT or MRI scan soon ,also do echocardiography and ultrasound to exclude other malformations.The treatment effect is satisfactory,and the outcome need long time to follow up.

Objective To evaluate the expression of metastasis-associated lung adenocarcinoma transcript 1 (MALAT-1) in ovarian cancer cell lines, and to investigate the biological effects of down-regulated MALAT-1 on OVCAR3 cells.Methods qRT-PCR analysis was used to examine the expression level of MALAT-1 gene in ovarian cancer cells, including ES-2, A2780, SKOV3 and OVCAR3 cell lines.For functional research, four shRNA oligos specially targeting MALAT-1 and a empty vector were designed and constructed into pGPU6/GFP/Neo, then transfected into OVCAR3 cells.qRT-PCR was used to confirm the effective suppression of MALAT-1.Changes of proliferation and adhesion of cells were analyzed by CCK-8 and adhesion assays.Wound-healing, transwell migration and invasion assays were used to examine migration and invasion of MALAT-l-silencing cells in vitro.Results The expression of MALAT-1 gene in OVCAR3 cells was high, and qRT-PCR results confirmed successfully the knockdown of MALAT-1 after transient transfection.After successful suppression of MALAT-1, the proliferation, wound-healing and adhesion ability in vitro were inhibited to some degree.In transwell migration assay, the number of migration cells in MALAT-1-silencing group was 52.17±4.48, which is much less than that in the negative and control groups (286.50± 12.23 and 295.67±6.96, respectively).In invasion assay, the number of invasion cells passing the transwell membrane in MALAT-1-silencing group (37.33±2.40) was also decreased significantly, compared to that in the negative and control groups (239.00±15.72 and 222.67±20.85, P ＜ 0.05).Conclusions shRNA-mediated silence of MALAT-1 can effectively inhibit the proliferation, adhesion, migration and invasion abilities of ovarian cancer cell line OVCAR3 in vitro, indicating MALAT-1 is expected to be a target gene for the treatment of ovarian cancer.

Objective To study the effect of bifidobacterium on intestinal tissue of necrotizing enterocolitis (NEC)in newborn rats and its regulation of Wnt/β-Catenin signal pathway.Methods Seventy -five newborn SD rats were randomly divided into 5 groups,and each group had 1 5 rats.Group A was artificial feeding control group;group B was NEC model group;group C was bifidobacterium treatment group;group D was artificial feeding +bifidobacterium control group;group E was rat breast feeding control group.The localization expression of Toll -like re-ceptor 4(TLR4)of ileocecal ileum tissue was detected by immunohistochemical detection,and also the equivalen-tileum tissues were detected for the contents of glycogen synthase kinase -3β(GSK3β)and β-Catenin expression by Wes-tern blot.Comparing the differences of these indicators between the groups,in addition,the data of TLR4,GSK3βandβ-Catenin were analyzed by Bivariate correlations.Results The levels of TLR4 in ileum tissue of 5 groups were 0.36 ±0.03,0.48 ±0.05,0.34 ±0.03,0.37 ±0.04,0.35 ±0.02.The levels of GSK3βin ileum tissue of 5 groups were 0.98 ±0.23,1 .48 ±0.42,0.99 ±0.20,0.56 ±0.1 7,0.60 ±0.1 5.The levels of β-Catenin in ileum tissue of 5 groups were 1 .48 ±0.22,0.64 ±0.55,1 .27 ±0.36,1 .72 ±0.51 ,1 .82 ±0.44.The levels of TLR4 and GSK3βin ileum tissue of group B were significantly increased compared with group E (P <0.05).The levels of β-Catenin sig-nificantly decreased compared with group E (P <0.05).The levels of TLR4 and GSK3βin ileum tissue of group C were significantly decreased compared with group B (P <0.05).The levels of β-Catenin significantly increased com-pared with group B (P <0.05).Negative correlation was observed between the levels of GSK3βand β-Catenin(r =-0.592,P <0.05),while positive correlation was observed between the levels of TLR4 and GSK3β(r =0.295,P <0.05),and negative correlation was observed between the levels of TLR4 and β-Catenin(r =-0.426,P <0.05). Conclusions Bifidobacterium has certain protective effect on the NEC newborn rat intestines,which can reduce the in-cidence of experimental NEC and the severity of intestinal injury.Its effect may be achieved by regulating the Wnt/β-Catenin signal pathway,which decreases the expression of the level of GSK3βand increases the level of repair fac-tor β-Catenin.

Objective:To analyze the effects of espO gene knockout on the biological characteristics of enterhemorrhagic Escherichia coli (EHEC). Methods:Two-step methods mediated by the suicide plasmid pCVD442-Δ espO and plasmid pTrc99a were used to construct the espO gene-deleted strain (Δ espO) and the complemented mutant (CΔ espO), respectively. HeLa cells were infected with different EHEC strains to analyze the biological functions and lethal effects of espO gene during infection. Results:PCR, electrophoresis and gene sequencing showed that the Δ espO and CΔ espO mutants were successfully constructed. Compared with the wild-type strain, neither the Δ espO nor CΔ espO mutant showed significant difference in growth rate, indicating that the espO gene had no influence on the growth and replication of EHEC. Furthermore, EspO could activate the tumor necrosis factor receptor (TNF)-induced NF-κB signaling pathway, while the effector protein NleB could inhibit the process. EspO could not inhibit the death of HeLa cells induced by TNF or TNF-related apoptosis-inducing ligand (TRAIL) after EHEC infection. Conclusions:In this study, we successfully constructed the espO gene-deleted and complemented mutants of EHEC and preliminarily analyzed the interaction between espO gene and host cells and the effects of espO gene on cell apoptosis during infection, which provided reference for further research on the in vitro biochemical activity and in vivo pathogenic roles of EspO.

Objective:To explore the factors related to functional recovery of the knee in patients with tibial plateau fracture complicated with intercondylar ridge fracture.Methods:Included in this retrospective study were 63 patients who had been treated at Emergency Center of Trauma, The Third Hospital of Hebei Medical University from January to June 2019 for tibial plateau fracture complicated with intercondylar ridge fracture. They were 43 males and 20 females, aged from 18 to 66 years (average, 43.9 years). According to our comprehensive classification of tibial plateau fractures, there were 27 simple ones (20 cases of type Ⅰ and 7 cases of type Ⅱ) and 36 complicated ones (16 cases of type Ⅲ, 17 cases of type Ⅳ, one of type Ⅴ and 2 cases of type Ⅵ). To screen out major influencing factors, a multiple linear regression model was used to analyze the 12 factors that might affect functional recovery of the knee by the Hospital for Special Surgery (HSS) scoring: age, gender, occupation, fracture type, way of medical payment, cause of injury, operation method, starting time for rehabilitation, postoperative brace installation, time from injury to operation, length of hospital stay, and presence or absence of a free intercondylar ridge fracture fragment.Results:The 63 patients were followed up for 6.0 to 7.1 months (average, 6.1 months). The HSS knee scores ranged from 45 to 100 points (average, 92.4 points). The HSS knee scores were significantly different between different fracture types ( P<0.05). The HSS scores were significantly higher for the patients without a free intercondylar ridge fracture fragment than for those with ( P<0.05). Multiple linear regression model analysis of the 5 variables with P<0.02 in the single factor analysis (age, fracture type, way of medical payment, hospital stay and presence or absence of a free intercondylar ridge fracture fragment) and HSS scores showed that only fracture type and presence or absence of a free intercondylar ridge fracture fragment had a significant impact on the knee function. Their regression equation was Y=125.591-7.790a-17.277b (Y indicates HSS score, a indicates fracture type and b indicates presence or absence of a free intercondylar ridge fracture fragment). Conclusions:The short-term prognosis for tibial plateau fractures of comprehensive types Ⅰ&Ⅱ (simple ones) may be better than that for tibial plateau fractures of comprehensive types Ⅲ, Ⅳ, Ⅴ&Ⅵ (complicated ones). For patients with a free intercondylar ridge fracture fragment on their lateral knee X-ray film, effective reduction and fixation is indicated but is not for those without.

OBJECTIVETo investigate the correlation between uncoupling protein 2 (UCP2) expression and myocardial mitochondria injury in rats with sepsis induced by lipopolysaccharide (LPS).

METHODSThe rat model of sepsis was established through an intraperitoneal injection of LPS. Forty male Sprague-Dawley rats were randomly and equally divided into control group (an intraperitoneal injection of normal saline), sepsis 6 h group (LPS-6 h group), sepsis 12 h group (LPS-12 h group), sepsis 24 h group (LPS-24 h group), and sepsis 48 h group (LPS-48 h group). The serum and heart tissues were harvested at corresponding time points and myocardial mitochondria was extracted. The microplate reader was applied to measure creatine kinase (CK), creatine kinase-MB (CK-MB), and reactive oxygen species (ROS). Flow cytometry was applied to measure the degree of mitochondrial swelling and mitochondrial membrane potential (MMP). Western blot was used to measure the expression level of UCP2. Electron microscopy was applied to observe the morphological changes in heart tissues and myocardial mitochondria.

RESULTSCompared with the control group, the LPS groups had significantly increased serum levels of CK, CK-MB, and myocardial ROS, as well as a significantly increased degree of mitochondrial swelling (P<0.05), and these values reached their peaks at 24 hours after LPS injection. The LPS groups had a significant decrease in MMP (P<0.05), which reached the lowest level at 24 hours after LPS injection. Western blot showed that the LPS groups had a significant increase in the expression level of myocardial UCP2 compared with the control group (P<0.05), which reached its peak at 24 hours after LPS injection. The results of electron microscopy showed mitochondrial swelling, partial rupture of the mitochondrial membrane, and cavity formation in rats in the LPS groups. The most severe lesions occurred in the LPS-24 h group. In rats with LPS, the ROS level in the myocardial mitochondria and the degree of mitochondrial swelling were positively correlated with the expression level of UCP2 (r=0.796 and 0.893, respectively; P<0.05), while MMP was negatively correlated with the expression level of UCP2 (r=-0.903, P<0.05).

CONCLUSIONSIn the rat model of sepsis, the myocardium and myocardial mitochondria have obvious injuries, and the expression level of UCP2 is closely correlated with mitochondrial injury. Therefore, UCP2 might play an important role in myocardial mitochondrial injury in sepsis.

Animals ; Cardiomyopathies ; genetics ; metabolism ; Disease Models, Animal ; Humans ; Ion Channels ; genetics ; metabolism ; Lipopolysaccharides ; adverse effects ; Male ; Mitochondria, Heart ; metabolism ; Mitochondrial Proteins ; genetics ; metabolism ; Myocardium ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sepsis ; genetics ; metabolism ; Uncoupling Protein 2

BACKGROUNDNeovascular glaucoma (NVG) is a refractory glaucoma. The management of NVG is very difficult, and it is more difficult when combined with vitreous hemorrhage. The aim of this study was to investigate the effects of ranibizumab plus combined surgery for NVG with vitreous hemorrhage.

METHODSA total of 26 eyes of 26 NVG patients with vitreous hemorrhage were recruited in this study. The patients aged from 36 to 63 years with a mean age of 51.97 ± 7.60 years. The mean intraocular pressure (IOP) was 46.38 ± 5.75 mmHg (1 mmHg = 0.133 kPa) while being treated with the maximum medical therapy. The mean best-corrected visual acuities converted to logarithm of the minimum angle of resolution (logMAR BCVA) was 2.62 ± 0.43. All the patients underwent intravitreal injection of 0.5 mg (0.05 ml) ranibizumab combined with pars plana vitrectomy (PPV), pars plana lensectomy (PPL) with a preserved anterior capsule, panretinal photocoagulation (PRP), and trabeculectomy (intravitreal ranibizumab [IVR] + PPV + PPL + PRP + trabeculectomy). The IOP and logMAR BCVA were the main outcome measures in this study.

RESULTSThe follow-up period was 12 months. The mean postoperative IOPs were 26.38 ± 3.75 mmHg, 21.36 ± 3.32 mmHg, 18.57 ± 3.21 mmHg, and 16.68 ± 2.96 mmHg, respectively at 7 days, 1 month, 3 months, and 12 months after PPV + PPL + PRP + trabeculectomy. At the last follow-up, the mean IOP was significantly lower than the preoperative one (t = 6.612, P = 0.001). At 7 days, 1 month, 3 months, and 12 months after PPV + PPL + PRP + trabeculectomy, the mean logMAR BCVA were 1.30 ± 0.36, 1.29 ± 0.37, 1.29 ± 0.39, and 1.26 ± 0.29, respectively. At the last follow-up, the mean logMAR BCVA was significantly improved, and the difference was statistically significant compared with preoperative one (t = 6.133, P = 0.002). The logMAR BCVA improved in 22 eyes (84.62%), and remained stable in 4 eyes (15.38%). The neovascularization in the iris and the angle regressed significantly in all patients 7 days after ranibizumab injection. No serious complications occurred during 12 months of the study.

CONCLUSIONSIVR + PPV + PPL + PRP + trabeculectomy can control IOP well and improve BCVA without severe complication for NVG patients with vitreous hemorrhage.

Adult ; Female ; Glaucoma, Neovascular ; drug therapy ; surgery ; Humans ; Intraocular Pressure ; drug effects ; Male ; Middle Aged ; Postoperative Complications ; Ranibizumab ; therapeutic use ; Trabeculectomy ; adverse effects ; Vitrectomy ; adverse effects ; Vitreous Hemorrhage ; drug therapy ; surgery

Objective:To evaluate the effect of epidural labor analgesia administration methods on occurrence of postpartum urinary retention in nulliparous parturients through a comparison between programmed intermittent epidural bolus (PIEB) and continuous epidural infusion.Methods:Two hundred nulliparous parturients who were at full term with a singleton fetus in vertex presentation, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, aged 22-35 yr, with body mass index of 22.4-42.6 kg/m 2, were divided into 2 groups ( n=100 each) using a random number table method: continuous epidural infusion group (group CEI) and group PIEB.Patient-controlled epidural analgesia (PCEA) was performed in active phase of labor (cervical dilatation≥1 cm) during the first stage of labor.The PCEA solution contained the mixture (10 ml) of 0.1% ropivacaine with 0.5 μg/ml sufentanil.The PCEA pump was set up to deliver a 5-ml bolus dose with a 30-min lockout interval.The analgesia solution contained the mixture (100 ml) of 0.08% ropivacaine and sufentanil 0.5 μg/ml.In group CEI, the drugs were given at 8 ml/h immediately after the initial dose.PIEB regimens were programmed as 8 ml over 80 s once an h after the initial bolus, and the administration was stopped after delivery of fetus.The labor time, consumption of analgesia solution and the incidence of postpartum urinary retention were recorded. Results:Compared with group CEI, the duration of the second stage of labor was significantly shortened, and consumption of analgesia solution and the incidence of postpartum urinary retention were decreased in group PIEB ( P<0.05). Conclusion:Compared with the continuous epidural infusion, the application of PIEB in labor analgesia can reduce the incidence of postpartum urinary retention in nulliparous parturients.