[Effect of SHP-1 knockout in airway epithelial cells on emphysema phenotype in chronic obstructive pulmonary disease in mice].

Objective: To construct and characterize conditional Src homology region 2 protein tyrosine phosphatase 1 (SHP-1) knockout mice in airway epithelial cells and to observe the effect of defective SHP-1 expression in airway epithelial cells on the emphysema phenotype in chronic obstructive pulmonary disease (COPD). Methods: To detect the expression of SHP-1 in the airway epithelium of COPD patients. CRISPR/Cas9 technology was used to construct SHP-1flox/flox transgenic mice, which were mated with airway epithelial Clara protein 10-cyclase recombinase and estrogen receptor fusion transgenic mice (CC10-CreER+/+), and after intraperitoneal injection of tamoxifen, airway epithelial SHP-1 knockout mice were obtained (SHP-1flox/floxCC10-CreER+/-, SHP-1Δ/Δ). Mouse tail and lung tissue DNA was extracted and PCR amplified to discriminate the genotype of the mice; the knockout effect of SHP-1 gene in airway epithelial cells was verified by qRT-PCR, Western blotting, and immunofluorescence. In addition, an emphysema mouse model was constructed using elastase to assess the severity of emphysema in each group of mice. Results: Airway epithelial SHP-1 was significantly downregulated in COPD patients. Genotyping confirmed that SHP-1Δ/Δ mice expressed CC10-CreER and SHP-1-flox. After tamoxifen induction, we demonstrated the absence of SHP-1 protein expression in airway epithelial cells of SHP-1Δ/Δ mice at the DNA, RNA, and protein levels, indicating that airway epithelial cell-specific SHP-1 knockout mice had been successfully constructed. In the emphysema animal model, SHP-1Δ/Δ mice had a more severe emphysema phenotype compared with the control group, which was manifested by disorganization of alveolar structure in lung tissue and rupture and fusion of alveolar walls to form pulmonary alveoli. Conclusions: The present study successfully established and characterized the SHP-1 knockout mouse model of airway epithelial cells, which provides a new experimental tool for the in-depth elucidation of the role of SHP-1 in the emphysema process of COPD and its mechanism.

[The prospect and challenge of liquid biopsy in the diagnosis and treatment of chest malignancy].

In recent years, the incidence of chest malignant tumors in China has increased year by year, which has seriously threatened the health problems of people. Among them, early screening and intervention of patients with chest malignancies is the key to cancer prevention. Early detection, early diagnosis, and early treatment as the "three early prevention" of clinical practice are conducive to improve the survival rate of tumor patients. As a non-invasive and real-time reflection of tumor status, liquid biopsy has gradually received attention in clinical diagnosis and treatment. Circulating tumor cells (CTCs), circulating tumor DNA (ctDNA) and exosomes as liquid biopsy "Three carriages" are not only widely used in the diagnosis, monitoring and prognostic evaluation of chest malignancies, but also face many unknown challenges. In this article, the application of liquid biopsy in chest malignancies in recent years is elaborated in detail, which provides a reference for the formulation of clinical tumor prevention and diagnosis and treatment strategies.

CTNNBIP1 modulates keratinocyte proliferation through promoting the transcription of ß-catenin/TCF complex downstream genes.

BACKGROUND: During psoriasis initiation and development, deregulations in signalling pathways and gene expression are observed. METHODS: Herein, we downloaded seven sets of microarray mRNA expression profiles showing differentially expressed genes in psoriasis lesion skin and non-lesion skin tissues and three sets of RNA-seq data and analysed these online data attempting to screen for crucial genes related to keratinocyte proliferation and psoriasis development. The expression of CTNNBIP1 in psoriasis patients and IMQ mouse model skin tissues were examined by RT-PCR, immunoblotting and IHC. The functions of CTNNBIP1 on HaCaT cell proliferation, apoptosis and ß-catenin/TCF complex were measured by MTT, EdU, flow cytometry, IF, luciferase assays and immunoblotting. RESULTS: The expression of catenin beta interacting protein 1 (CTNNBIP1) was remarkably downregulated within psoriasis lesion skin tissue samples compared to that within non-lesion skin tissues based on both online data and experimental results. In response to a period of different therapies, respectively, CTNNBIP1 expression could be rescued in lesion skin tissues. Within IMQ-induced psoriasis-like dermatitis in mice, CTNNBIP1 silence further aggravated psoriatic phenotypes. In human immortalized keratinocytes, HaCaT cells, CTNNBIP1 silence significantly inhibited cell apoptosis and promoted cell proliferation. Regarding the molecular mechanism, CTNNBIP1 silence in HaCaT cells promoted ß-catenin nucleus translocation, enhanced the transcriptional activity of TCF4 and increased ß-catenin/TCF complex downstream c-Myc and cyclin D1 proteins, and also increased the expression of cell proliferation marker ki-67. In contrast to CTNNBIP1, the expression of c-Myc and cyclin D1 showed to be dramatically upregulated within psoriasis lesion tissue samples than that within non-lesion tissue samples. Within tissues, c-Myc and cyclin D1 showed to be negatively correlated with CTNNBIP1, respectively. CONCLUSIONS: We identify CTNNBIP1 as an abnormally downregulated gene in psoriasis. CTNNBIP1 silence significantly disturbs the proliferation of keratinocytes through promoting the transcription of ß-catenin/TCF complex downstream genes.

Prevalence of positivity to antibodies to hepatitis C virus among volunteer blood donors in China: a meta-analysis.

OBJECTIVES: Safe blood transfusion plays an important role in the prevention of transfusion-transmissible infections, and hepatitis C virus (HCV) infection is one of the major problems associated with this procedure. This meta-analysis aimed to determine the prevalence of HCV infection in Chinese blood donors. STUDY DESIGN: The study design of this study is a meta-analysis. METHODS: Eligible studies were retrieved from PubMed, Embase, China National Knowledge Infrastructure, China Science and Technology Journal Database and Wanfang literature databases from 2010 to 2020. The effect measure was presented as HCV prevalence with a 95% confidence interval (CI). Q test was used to assess the heterogeneity, and the I2 statistics was determined to decide whether a random effects model or a fixed effects model should be used as the pooling method. Subgroup analyses were also conducted. RESULTS: A total of 62 eligible studies, including 9,007,220 HCV blood donors, were analysed. Of the total blood donors, 35,017 were infected with HCV. The pooled HCV prevalence was 0.415% (95% CI: 0.371-0.458). The subgroup analysis revealed that the prevalence of positivity to anti-HCV antibodies was significantly different in each year (P < 0.05). However, no significant difference was observed in HCV prevalence in terms of sex. Moreover, the prevalence of positivity to anti-HCV was remarkably higher in first-time blood donors than in repeat blood donors (P < 0.05), and the rate of HCV infection among university students was significantly lower than that among soldiers (P < 0.05). CONCLUSIONS: The rate of HCV infection showed a downward trend from 2010 to 2014, increased in 2015-2016, and finally decreased in 2017-2018. Thus, the prevalence of HCV infection has decreased in Chinese blood donors after comprehensive prevention and treatment.

[Study on the expression of p62 protein in patients with lung adenocarcinoma].

To explore the expression of p62 protein in lung adenocarcinoma (LUAD). In this study, a cross-sectional study was adopted. From December 2011 to May 2013, 60 patients with lung adenocarcinoma who were diagnosed and treated in Tongji Hospital of Tongji University, Shanghai were selected for paraffin embedding and tissue chip preparation, and immunohistochemistry (IHC) technology was used to detect the expression of p62 in lung adenocarcinoma patients' cancer tissues and adjacent tissues, and analyze the relationship between p62 expression and the clinicopathological characteristics and survival prognosis of patients with lung adenocarcinoma; at the same time, 6 cases of lung adenocarcinoma were selected by random sampling cancer tissues and adjacent tissues were detected by Western Blot (WB) to detect p62 protein and analyzed by gray value. Preoperative examination specimens of inpatients with lung adenocarcinoma diagnosed from April 2018 to early October 2019, and plasma specimens of healthy subjects were collected, and enzyme linked immunosorbent assay (ELISA) was used to detect lung adenocarcinoma patients and healthy patients. The expression of p62 in the plasma of the subjects was statistically analyzed using SPSS 22.0 software. The results of IHC showed that the positive expression rate of p62 in cancer tissues was significantly higher than that in adjacent tissues, and the difference was statistically significant (t=5.593, P<0.001). Similarly, WB results showed that the expression of p62 protein in cancer tissues was significantly higher than that in adjacent tissues. It is statistically relevant (t=2.238, P=0.049). The expression of p62 was statistically correlated with tumor size, clinicopathological stage and lymph node metastasis in patients with lung adenocarcinoma (all P<0.05). The overall survival of patients with lung adenocarcinoma with high p62 expression was worse than that of patients with low p62 expression (95%CI was 0.238-0.870, P=0.028), suggesting that the high expression of p62 is related to the poor prognosis of patients with lung adenocarcinoma. The level of p62 protein in the plasma of patients with lung adenocarcinoma was significantly higher than that in the healthy control group. The difference was statistically significant (t=8.533, P<0.001). The area under the receiver operating characteristic curve was 0.835 (95%CI was 0.779-0.891, P<0.001), which is significantly higher than CEA, CA125, CA153 and other single traditional indicators, and the combined detection of four indicators has the highest diagnostic efficiency. p62 was strongly expressed in cancer tissues and serum, which is related to the poor prognosis and overall survival rate of LUAD patients.

[Safety of biological valves for aortic valve replacement: A systematic review and meta-analysis].

OBJECTIVE: To provide a comprehensive and contemporary overview of the long-term safety outcomes after aortic valve replacements (AVR) with conventional biological heart valve (stented or stentless). METHODS: English databases (Medline, Embase, Web of Science, CENTRAL, and ClinicalTrial.gov) and Chinese databases (CNKI, VIP, WanFang, and SinoMed) were searched systemically from January 1, 2000 to January 26, 2019. Eligible randomized controlled trials, non-randomized clinical trials, cohort studies (retrospective or prospective), and unselected case series were included. Strict screening of the obtained literature was conducted to extract relevant data by two reviewers. Other inclusion criteria were studied reporting on outcomes of AVR with biological valves (stented or stentless), with or without coronary artery bypass grafting (CABG) or valve repair procedure, with mean follow-up length equal to or longer than 5 years. We excluded studies that reported only a specific patient group (e.g., patients with renal failure, or pregnancy), without the report of biological valve type, or with study population size less than 100. The meta-analysis was performed using Stata 14.0 software. RESULTS: In this study, 53 papers (in total 57 study groups) involving 47 803 patients were included. (1) The all-cause mortality was 6.33/100 patient-years (95%CI: 5.85-6.84). Subgroup analysis showed that the mortality rates of porcine and bovine valve prostheses were 5.69/100 patient-years (95%CI: 5.05-6.41) and 7.29/100 patient-years (95%CI: 6.53-8.13), respectively. The all-cause mortality rates for stented and stentless valve were 6.69/100 patient-years (95%CI: 6.12-7.30) and 5.21/100 patient-years (95%CI: 4.43-6.14), respectively. (2) The incidence rate of thromboembolism was 1.16/100 patient-years (95%CI: 0.96-1.40), the incidence rate of permanent pacemaker (PPM) implantation was 1.08/100 patient-years (95%CI: 0.75-1.54), the incidence rate of stroke was 0.74/100 patient-years (95%CI: 0.51-1.06), the incidence rate of structural valve dysfunction (SVD) was 0.73/100 patient-years (95%CI: 0.59-0.91), the incidence rate of major bleeding was 0.52/100 patient-years (95%CI: 0.41-0.65), the incidence rate of endocarditis was 0.38/100 patient-years (95%CI: 0.33-0.44), and the incidence rate of non-structural valve dysfunction (NSVD) was 0.20/100 patient-years (95%CI: 0.13-0.31). The total reoperation rate for biological aortic valve was 0.77/100 patient-years (95%CI: 0.65-0.91), and the SVD related reoperation rate was 0.46/100 patient-years (95%CI: 0.36-0.58). CONCLUSION: The all-cause mortality for conventional biological AVR was 6.33/100 patient-years. Thromboembolism, PPM implantation, reoperation, stroke, and SVD were major long term complications.

[Cytogenetic abnormalities and morphological changes of bone marrow in multiple myeloma: a pathological analysis of 151 cases].

Objective: To investigate the relationship between six common cytogenetic abnormalities and bone marrow pathomorphology in multiple myeloma (MM). Methods: Bone marrow biopsy was performed on 151 newly-diagnosed MM patients. Meanwhile, myeloma cells were enriched by CD138 immunomagnetic beads, and then lq+, 13q-, 17p-, t(4;14), t (11;14), t (14;16) and other common genetic abnormalities were detected using interphase fluorescence in situ hybridization (FISH). The relationship between different genetic abnormalities and biopsy morphology was compared. Results: Of the 151 patients, 15 had extramedullary infiltration (9.9%). The rate of cytogenetic abnormalities was 76.2% (115/151), of which 1q+ accounted for 49.7% (75/151), 13q-39.1% (59/151), 17p-8.6% (13/151), t(4;14) 21.2% (32/151), t(11;14) 19.2% (29/151), and t(14;16) 2.0% (3/151). The proliferation patterns of MM plasma cells were nodular (48.3%, 73/151), interstitial (33.8%, 51/151) and diffuse (17.9%, 27/151). The morphology of plasma cells was mainly mature type (58.3%, 88/151), followed by juvenile type (20.5%, 31/151), intermediate type (15.9%, 24/151) and plasmacyte type (5.3%, 8/151). According to the mSMART risk stratification system, the proliferation pattern of myeloma cells in the high-risk group was mainly diffuse type, and the morphology was mainly immature and plasmacyte type. In the middle-risk group, mature type myeloma cells were mainly nodular proliferating. In the low-risk and negative group, mature type myeloma cells were mainly interstitial proliferating. There was no difference in the probability of different proliferation modes of intermediate type plasma cells in each group. Conclusions: The proliferation pattern and morphology of plasma cells in bone marrow biopsy combined with cytogenetic markers can more accurately predict the severity and prognosis of MM.

Effects of Moringa oleifera silage on milk yield, nutrient digestibility and serum biochemical indexes of lactating dairy cows.

This study investigated the effects of Moringa oleifera (MO) as a partial substitute of alfalfa hay on milk yield, nutrient apparent digestibility and serum biochemical indexes of dairy cows. MO was harvested at 120 days post-seeding. Fresh MO was cut, mixed with chopped oat hay (425:575 on a DM basis), ensiled and stored for 60 days. Sixty healthy Holstein dairy cows were allocated to one of three groups: NM (no MO or control), LM (low MO; 25% alfalfa hay and 50% maize silage were replaced by MO silage) or HM (high MO; 50% alfalfa hay and 100% maize silage were replaced by MO silage). The feeding trial lasted 35 days. The LM and HM diets did not affect dry matter (DM) intake, milk yield or milk composition (lactose, milk fat, milk protein and somatic cell count). The apparent digestibility of DM and NDF was lower for HM group than NM group. Additionally, there were no significant differences in serum biochemical indexes between the LM and NM groups. The HM group had lower serum concentrations of total cholesterol, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol and higher serum concentrations of urea than the NM group. The partial replacement of alfalfa hay (≤50%) and maize silage with MO silage had no negative effects on milk yield, in vivo nutrient apparent digestibility or serum biochemical indexes of lactating cows.

[Effect of the vacuum-formed retainer on preventing the proximal contact loss between implant supported crown and adjacent natural teeth].

OBJECTIVE: To evaluate the effect of the vacuum-formed retainer on preventing the proximal contact loss between the implant supported crown and its adjacent natural teeth. METHODS: Forty-six posterior implant crowns in the mandible including 92 interproximal contacts in 46 patients (19 men, 27 women) aged from 25 to 66 years were included. The participants in experimental group (22 cases) were vacuum-formed retainers at night, while participants in control group (24 cases) only received routine examination. The two groups were not different in age, gender, the time interval of the tooth loss and tooth position at baseline. Mesial and distal proximal contact tightness was measured using the orthodontic dynamometer and metallic articulating film immediately after crown delivery, and 1-month, 3-month, 6-month, and 1-year follow-up respectively. The articulating film was inserted interdentally from the occlusal direction, and then it was slowly removed in the buccallingual direction by the dynamometer. Increasing the number of films (N) piece by piece until the frictional force (F) was great than 0, and the number of films (N) was recorded. At each follow-up, proximal contact between implant crown and its adjacent teeth was considered to be loss if the number of films (N) used at immediate crown delivery passed without frictional force (F=0). Besides, the periodontal conditions [scored according to the probing depth (PD), bleeding index (BI), mobility (M)] and complaint of food impaction were recorded. The mesial and distal proximal contact loss rates were compared between the two groups at different times. Chi-square test or Fisher's exact test was used for statistical analysis. RESULTS: The proximal contact loss rate on the mesial surface of the implant supported crown continuously increased over the follow-up periods. At the end of the 1-, 3-, and 6-month follow ups, 18.2%, 22.7% and 27.3% were identified for the contact loss rates on the mesial surface of the implant supported crown in the experimental group, respectively. Meanwhile in control group, the rates were 20.8%, 37.5% and 45.8%. No significant differences were observed at the end of the 1-, 3-, and 6-month follow ups(1-month: χ2=0.000, P=1.000; 3-month: χ2=1.183, P=0.277; 6-month: χ2=1.697, P=0.193). The proximal contact loss rate on the mesial surface in control group (62.5%) was significantly higher than that in the experimental group (31.8%, χ2=4.330, P=0.037) at the end of the 1-year follow-up. However, no statistical difference was found on the distal surfaces between the two groups during the whole follow-up periods. The first open contact was noted 1 month after crown insertion. CONCLUSION: By wearing vacuum-formed retainer for one year, the incidence of open contacts between the posterior implant prostheses and mesial adjacent teeth in the mandible has been reduced.

Gut microbiome remodeling induces depressive-like behaviors through a pathway mediated by the host's metabolism.

Major depressive disorder (MDD) is the result of complex gene-environment interactions. According to the World Health Organization, MDD is the leading cause of disability worldwide, and it is a major contributor to the overall global burden of disease. However, the definitive environmental mechanisms underlying the pathophysiology of MDD remain elusive. The gut microbiome is an increasingly recognized environmental factor that can shape the brain through the microbiota-gut-brain axis. We show here that the absence of gut microbiota in germ-free (GF) mice resulted in decreased immobility time in the forced swimming test relative to conventionally raised healthy control mice. Moreover, from clinical sampling, the gut microbiotic compositions of MDD patients and healthy controls were significantly different with MDD patients characterized by significant changes in the relative abundance of Firmicutes, Actinobacteria and Bacteroidetes. Fecal microbiota transplantation of GF mice with 'depression microbiota' derived from MDD patients resulted in depression-like behaviors compared with colonization with 'healthy microbiota' derived from healthy control individuals. Mice harboring 'depression microbiota' primarily exhibited disturbances of microbial genes and host metabolites involved in carbohydrate and amino acid metabolism. This study demonstrates that dysbiosis of the gut microbiome may have a causal role in the development of depressive-like behaviors, in a pathway that is mediated through the host's metabolism.

Intestinal microbiota could transfer host Gut characteristics from pigs to mice.

BACKGROUND: The present study was conducted to compare the differences in gut microbiota composition and gut-phenotypes among pig breeds, and determine whether these differences would transmit to mice colonized with fecal microbiota of different pig breeds. A total of 24 1-day-old germ-free BALB/C mice were divided into 3 groups (TFM, YFM and RFM), which were transplanted with intact fecal microbiota of Tibetan pig (TP), Yorkshire pig (YP) and Rongchang pig (RP), respectively. RESULTS: Results showed that different pig breeds exhibited distinct gut microbiota profile based on high-throughput pyrosequencing. YP exhibited a lower Firmicutes/Bacteroidetes ratio and apparent genera differences compared with RP and TP, and higher levels of bacteria from Spirochaetes were observed in TP compared with RP and YP (P < 0.05). Transplanted porcine microbiota into GF mice replicated the phenotypes of pig donors. Moreover, the three groups of donor pigs and their mice recipients exhibited different intestinal index and morphology. TP and RP had higher intestinal weight and relative CDX2 mRNA expression in ileum than YP, and longer intestine, higher villus height of duodenum and jejunum were observed in TP compared with YP and RP (P < 0.05). TP exhibited higher GLP-2 mRNA expression in duodenum and jejunum than RP (P < 0.05). Similarly, YFM had lower intestine weight and CDX2 mRNA expression in ileum than TFM and RFM (P < 0.05). The intestine length in TFM was longer than that in RFM, and TFM had higher villus height in duodenum and jejunum and GLP-2 mRNA expression in ileum than the other two groups (P < 0.05). Besides, the digestive and absorptive ability was different among the three groups in donor pigs and mice recipients. YP had higher jejunal lactase and maltase activities than TP and RP, while TP had higher activities of jejunal ATPase, γ-GT, and relative SGLT1 mRNA expression in duodenum and jejunum than YP and RP (P < 0.05). Likewise, YFM had higher jejunal sucrase and maltase activities than TFM and RFM, whereas higher jejunal γ-GT activity and relative SGLT1 mRNA expression in duodenum and ileum were observed in TFM compared with YFM and RFM (P < 0.05). In addition, Tibetan pigs-derived microbiota improved gut barrier in mice recipients. The concentration of MDA in YP was higher than that in TP and RP (P = 0.078), and the relative ZO-1 mRNA expression in ileum in TP was higher than that in YP (P < 0.05). Likely, compared with TFM and RFM, YFM exhibited increasing MDA concentration in jejunum (P = 0.098), and the relative ZO-1 mRNA expression in duodenum and ileum in TFM were higher than that in YFM (P < 0.05). CONCLUSIONS: There were huge differences in gut microbiota composition and gut characteristics among pig breeds, and gut microbiota could partially convey host gut characteristics from pigs to mice.

Orthogonal design in the optimization of a start codon targeted (SCoT) PCR system in Roegneria kamoji Ohwi.

Roegneria kamoji Ohwi is an excellent forage grass due to its high feeding value and high resistance to some biotic and abiotic stresses. However, the start codon targeted (SCoT) polymorphism has not been conducted on R. kamoji. In this study, an orthogonal L16 (45) design was employed to investigate the effects of five factors (Mg2+, dNTPs, Taq DNA polymerase, primer, and template DNA) on the polymerase chain reaction (PCR) to determine the optimal SCoT-PCR system for R. kamoji. The results showed that the most suitable conditions for SCoT-PCR in R. kamoji included 1.5 mM Mg2+, 0.15 mM dNTPs, 1.0 U Taq DNA polymerase, 0.4 pM primer, and 40 ng template DNA. SCoT primers 39 and 41 were used to verify the stability of the optimal reaction system, and amplification bands obtained from diverse samples were found to be clear, rich, and stable in polymorphisms, indicating that this reaction system can be used for SCoT-PCR analysis of R. kamoji. We have developed a simple and rapid way to study the mutual effects of factors and to obtain positive results through the use of an orthogonal design L16 (45) to optimize the SCoT-PCR system. This method may provide basic information for molecular marker-assisted breeding and analyses of genetic diversity in R. kamoji.

Optimizing broadband terahertz modulation with hybrid graphene/metasurface structures.

We demonstrate efficient terahertz (THz) modulation by coupling graphene strongly with a broadband THz metasurface device. This THz metasurface, made of periodic gold slit arrays, shows near unity broadband transmission, which arises from coherent radiation of the enhanced local-field in the slits. Utilizing graphene as an active load with tunable conductivity, we can significantly modify the local-field enhancement and strongly modulate the THz wave transmission. This hybrid device also provides a new platform for future nonlinear THz spectroscopy study of graphene.

[Application of standardization of medical care model for chronic mild lead poisoning].

OBJECTIVE: To standardize the treatment and evaluate the clinical application effectiveness of clinical pathway (CP) in chronic mild lead poisoning. METHODS: 60 patients with chronic mild lead poisoning hospitalized from Jan. 2014 to Dec.2014 were enrolled for the study group, 60 patients with chronic mild lead poisoning hospitalized from Jan.2013 to Dec.2013 were enrolled for the control group. The study group were cared according to clinical pathway, the control group received routine therapy; the clinical application effectiveness were compared between the two groups. RESULTS: No adverse drug reactions occurred in both groups. The curative ratio was significantly higher in the study group than in the control group (96.7% vs 86.0%, P<0.05) , the rate of patients' satisfaction to medical care significantly higher in the study group (98.3% vs 88.3%, P<0.05) , the rate of health education awareness higher in the study group (95.0% vs 81.7%, P<0.05) , the course of treatment shorter in the study group (3.2±0.6 vs 3.4±0.7, P<0.05) , the medical cost less in the study group (5773.5 yuan±1242.1 yuan vs 6354.7 yuan±1177.0 yuan, P<0.05) , the length of hospitalization was shorter in the study group (21.9 d±6.7 d vs 24.6 d±7.9 d, P<0.05). The variation rate of clinical pathway was 13.3% in clinical pathway group. CONCLUSION: The implementation of clinical pathway could improve the curative ratio, satisfaction, and health education awareness. The course of treatment, length of hospital stay and costs of hospitalization in the study group could be obviously shorter and less, and there is a little variation rate in the clinical pathway.

Transcription factor CAAT/enhancer-binding protein is involved in regulation of expression of sterol carrier protein x in Spodoptera litura.

The Spodoptera litura sterol carrier protein x (SlSCPx) gene is expressed in various tissues throughout the life cycle and plays important role in sterol absorption and transport. In this study, the effects of insect hormones (20-hydroexcdysone and juvenile hormone) and lipids (arachidonic acid, cholesterol) on the expression of SlSCPx was analysed by reverse-transcriptase PCR. The results showed that none of these substances significantly induced the expression of SlSCPx in Spodoptera litura-221 (Spli-221) cells. To identify the transcription factors responsible for regulation of SlSCPx expression, a 3311-bp promoter sequence of the gene was cloned. Transcriptional activity of the promoter was studied using an in vivo promoter/reporter system and a 29-bp sequence between -1000 and -1029 nucleotides (nt) upstream of this gene was found to be responsible for the up-regulation of the gene. Over-expression of CAAT/enhancer-binding protein (C/EBP) in Spli-221 cells increased the promoter activity 5.57-fold. An electrophoretic mobility shift assay showed that two nuclear proteins bound to this sequence. Recombinant C/EBP specifically bound with a putative cis-regulatory element (CRE). Mutation of the C/EBP CRE abolished the binding of the C/EBP with the CRE. These results suggest that the transcription factor C/EBP may regulate the expression of SlSCPx by binding to the CRE in the promoter of this gene.

Transgenic characterization of two testis-specific promoters in the silkworm, Bombyx mori.

Sex-specific regulatory elements are key components for developing insect genetic sexing systems. The current insect genetic sexing system mainly uses a female-specific modification system whereas little success was reported on male-specific genetic modification. In the silkworm Bombyx mori, a lepidopteran model insect with economic importance, a transgene-based, female-specific lethality system has been established based on sex-specific alternative splicing factors and a female-specific promoter BmVgp (vitellogenin promoter) has been identified. However, no male-specific regulatory elements have yet been identified. Here we report the transgenic identification of two promoters that drive reporter gene expression in a testis-specific manner in B. mori. Putative promoter sequences from the B. mori Radial spoke head 1 gene (BmR1) and beta-tubulin 4 gene (Bmß4) were introduced using piggybac-based germline transformation. In transgenic silkworms, expression of the reporter gene enhanced green fluorescent protein (EGFP) directed by either BmR1 promoter (BmR1p) or Bmß4p showed precisely testis-specific manners from the larval to adult stage. Furthermore, EGFP expression of these two transgenic lines showed different localization in the testis, indicating that BmR1p or Bmß4p might be used as distinct regulatory elements in directing testis-specific gene expression. Identification of these testis-specific promoters not only contributes to a better understanding of testis-specific gene function in insects, but also has potential applications in sterile insect techniques for pest management.

Protein-responsive assemblies from catechol-metal ion supramolecular coordination.

Supramolecular self-assembly driven by catechol-metal ion coordination has gained great success in the fabrication of functional materials including adhesives, capsules, coatings and hydrogels. However, this route has encountered a great challenge in the construction of nanoarchitectures in the absence of removable templates, because of the uncontrollable crosslinking of catechol-metal ion coordination. Herein, we show that a supramolecular approach, combining both catechol-metal ion coordination and polymer self-assembly together, can organize polymers into hybrid nanoassemblies ranging from solid particles, homogeneous vesicles to Janus vesicles. Without the introduction of a specific binding ligand or complicated molecular design, these assemblies can totally disassemble in response to proteins. UV/vis absorption, fluorescence quenching and recovery investigations have confirmed that proteins can seize metal ions from the hybrid nanoassemblies, thus causing the degradation of catechol-metal ion coordination networks.

Novel EDA mutation in X-linked hypohidrotic ectodermal dysplasia and genotype-phenotype correlation.

OBJECTIVES: X-linked hypohidrotic ectodermal dysplasia (XLHED) is characterized by abnormalities of hair, teeth, and sweat glands, while non-syndromic hypodontia (NSH) affects only teeth. Mutations in Ectodysplasin A (EDA) underlie both XLHED and NSH. This study investigated the genetic causes of six hypohidrotic ectodermal dysplasia (HED) patients and genotype-phenotype correlation. METHODS: The EDA gene of six patients with HED was sequenced. Bioinformatics analysis and structural modeling for the mutations were performed. The records of 134 patients with XLHED and EDA-related NSH regarding numbers of missing permanent teeth from this study and 20 articles were reviewed. Nonparametric tests were used to analyze genotype-phenotype correlations. RESULTS: In four of the six patients, we identified a novel mutation c.852T>G (p.Phe284Leu) and three reported mutations: c.467G>A (p.Arg156His), c.776C>A (p.Ala259Glu), and c.871G>A (p.Gly291Arg). They were predicted to be pathogenic by bioinformatics analysis and structural modeling. Genotype-phenotype correlation analysis revealed that truncating mutations were associated with more missing teeth. Missense mutations and the mutations affecting the TNF homology domain were correlated with fewer missing teeth. CONCLUSIONS: This study extended the mutation spectrum of XLHED and revealed the relationship between genotype and the number of missing permanent teeth.

Optimization of SCoT-PCR reaction system in Dactylis glomerata by orthogonal design.

The effects of 5 factors (template DNA, Mg(2+), dNTPs, Taq DNA polymerase, and primer) on the polymerase chain reaction (PCR) were investigated to optimize the start codon targeted polymor-phism (SCoT)-PCR system of Dactylis glomerata L., using an orthogo-nal design L16 (4(5)). A suitable SCoT-PCR system for D. glomerata was established; the 20 µL reaction volume contained 3.0 mM Mg(2+), 0.2 mM dNTPs, 1.0 U Taq DNA polymerase, 0.2 µM primer, 20 ng tem-plate DNA, and 2 µL 10X buffer. Each factor had a different effect on the amplification reaction, and the concentration of dNTPs had the larg-est effect on the SCoT-PCR system. We tested 10 orchardgrass samples to determine and verify the stability of the reaction system. The results showed that amplified bands from diverse materials were clear, stable, and rich in polymorphisms, indicating that the optimized system was very stable.

Controlling graphene ultrafast hot carrier response from metal-like to semiconductor-like by electrostatic gating.

We investigate the ultrafast terahertz response of electrostatically gated graphene upon optical excitation. We observe that the photoinduced terahertz absorption increases in charge neutral graphene but decreases in highly doped graphene. We show that this transition from semiconductor-like to metal-like response is unique for zero bandgap materials such as graphene. In charge neutral graphene photoexcited hot carriers effectively increase electron and hole densities and increase the conductivity. In highly doped graphene, however, photoexcitation does not change net conducting carrier concentration. Instead, it mainly increases electron scattering rate and reduce the conductivity.