[Analysis of single-nucleotide polymorphism of Sonic hedgehog signaling pathway in non-syndromic cleft lip and/or palate in the Chinese population].

OBJECTIVE: To study the relationship between Sonic hedgehog (Shh) associated single-nucleotide polymorphism (SNP) and non-syndromic cleft lip and/or palate (NSCL/P), and to explore the risk factors of cleft lip and/or palate. Many studies suggest that the pathogenesis of NSCL/P could be related to genes that control early development, in which the Shh signaling pathway plays an important role. METHODS: Peripheral blood was collected from 197 individuals (100 patients with NSCL/P and 97 healthy controls). Haploview software was used for haplotype analysis and Tag SNP were selected, based on the population data of Han Chinese in Beijing of the international human genome haplotype mapping project. A total of 27 SNP were selected for the 4 candidate genes of SHH, PTCH1, SMO and GLI2 in the Shh signaling pathway. The genotypes of 27 SNP were detected and analyzed by Sequenom mass spectrometry. The data were analyzed by chi-squared test and an unconditional Logistic regression model. RESULTS: The selected SNP basically covered the potential functional SNP of the target genes, and its minimum allele frequency (MAF) was >0.05: GLI2 73.5%, PTCH1 91.0%, SMO 100.0%, and SHH 75.0%. It was found that the genotype frequency of SNP (rs12674259) located in SMO gene and SNP (rs2066836) located in PTCH1 gene were significantly different between the NSCL/P group and the control group. Linkage disequilibrium was also found on 3 chromosomes (chromosomes 2, 7 and 9) where the 4 candidate genes were located. However, in the analysis of linkage imbalance haplotype, there was no significant difference between the disease group and the control group. CONCLUSION: In China, NSCL/P is the most common congenital disease in orofacial region. However, as it is a multigenic disease and could be affected by multiple factors, such as the external environment, the etiology of NSCL/P has not been clearly defined. This study indicates that Shh signaling pathway is involved in the occurrence of NSCL/P, and some special SNP of key genes in this pathway are related to cleft lip and/or palate, which provides a new direction for the etiology research of NSCL/P and may provide help for the early screening and risk prediction of NSCL/P.

[Laparoscopy and endoscopy cooperative surgery in the treatment of stomach neoplasms].

With the rapid development of medical technology and the improvement of people's health awareness, the detection rate of benign gastric tumors and early gastric cancer has increased significantly. Under the premise of ensuring the safety of oncology, challenges for surgeons present is how to adopt precise and reasonable treatment plans according to the characteristics of gastric tumors to minimize surgical trauma and complications, improve postoperative quality of life, and achieve individualized and precise treatment. Laparoscopic surgery and digestive endoscopy are currently the two main methods for treating gastric tumors. However, they both have advantages and shortcomings. The combination of laparoscopy and digestive endoscopy for the treatment of gastric tumors has become a new way to treat gastric tumors. This operation not only fully exploits the advantages of laparoscopy and digestive endoscopy, but also complements the shortcomings of each. This article reviews the surgical technique categories, indications, technical improvements, and perspectives of laparoscopy combined with digestive endoscopy in the treatment of gastric tumors.

Vertical microbiological variation of a coastal aquifer in southern China.

Microbial communities in a coastal aquifer in the Zhuhai region of southern China were investigated by culture-independent molecular approaches. Four 16S rRNA gene libraries of three groundwater samples from varied depths and one seawater sample were constructed and analysed by the amplified ribosomal DNA restriction analysis technique (ARDRA). The phylogenetic analysis indicated that the 16S rDNA of clones presenting dominant ARDRA patterns were most similar to Proteobacteria, Bacteroidetes, Actinobacteria, Planctomycetes, and candidate divisions OPx (such as OP3, OP8, and OP11). In samples extracted from wells of 5-, 20-, and 35-m depth (i.e., D1, D5, and D6) Proteobacteria made up 32.3, 34.3, and 46.7% of the microbial communities, respectively. The same samples from D1, D5, and D6 also consisted of 5.0, 11.2, and 6.5% Bacteroidetes and 5.4, 6.6, and 7.8% Actinobacteria, respectively. In contrast, the seawater clone library had a predominant number of Proteobacteria (32.8%), while Bacteroidetes and Planctomycetes both accounted for 9.3%. Total microbial diversity remained relatively constant over the top layer to a depth of approximately 35 m, significant community vertical and horizontal (seawater-groundwater) shifts were observed for certain bacterial populations.

[Pelvic membrane anatomy and surgery with network preservation of autonomic nervous system for rectal cancer].

The principle of total mesorectal excision (TME) standardizes the resection range and surgical dissection plane in radical rectal cancer surgery, reduces the local recurrence rate and improves the long-term survival. TME is the "gold standard" in radical rectal cancer surgery. However, with the progress of laparoscopic surgical instruments and techniques in recent years, further understanding of pelvic membrane anatomy and autonomic nervous system has been gained, which makes the surgical plane of TME more accurate and the autonomic nervous system better preserved. According to anatomical discovery and histological confirmation, there is a fascia between the mesorectal fascia and pelvic parietal fascia, called pre-hypogastric nerve sheath, in which autonomic nervous system courses, including the superior hypogastric plexus, left and right hypogastric nerves, pelvic plexus and the neurovascular bundles, from the abdominal to the pelvic cavity behind the mesorectal fascia. It fuses with the end of the mesorectum at the superior border of musculi puborectalis, and goes around the mesorectum to join with Denonvillier fascia. On the basis of anatomical studies and empirical anatomical observations, we put forward the concept of network preservation of the autonomic nervous system: the main trunk as well as the nerve branches of the pelvic autonomic nervous system and accompanying blood vessels should be preserved to ensure the integrity of the nerve reflex arc. The concept allows the radical resection of rectal cancer to follow the principle of TME, and meanwhile, protect patient's urination function and sexual function to the greatest extent, improving the quality of life of patients after surgery.

Sensitive and rapid detection of Vibrio corallilyticus by loop-mediated isothermal amplification targeted to the alpha subunit gene of RNA polymerase.

AIMS: A diagnostic protocol was developed for rapid detection of Vibrio corallilyticus by method of loop-mediated isothermal amplification (LAMP). METHODS AND RESULTS: For cloning and sequencing of rpo A gene of V. corallilyticus, a set of four LAMP primers were designed by targeting the rpoA gene. With Bst DNA polymerase, the reaction time and temperature were optimized for 70 min at 65 degrees C, respectively. The amplification products were detected by electrophoresis. The detection limit of V. corallilyticus by LAMP was 3.6 x 10(3) CFU ml(-1) (8 CFU per reaction), but PCR could detect up to 3.6 x 10(4) CFU ml(-1) (72 CFU per reaction). The LAMP method was ninefold more sensitive than conventional PCR. The results also indicated that the LAMP reaction was highly specific to V. corallilyticus. CONCLUSIONS: The LAMP assay was a sensitive, specific and cost-effective method for the rapid detection of V. corallilyticus. SIGNIFICANCE AND IMPACT OF THE STUDY: This LAMP method provides an important diagnostic tool for the detection of V. corallilyticus infection. It can replace laborious biochemical tests for the identification of V. corallilyticus.

[Feasibility, safety and long-term efficacy of laparoscopic total gastrectomy combined with distal pancreaticosplenectomy for T4b gastric cancer].

Objective: To explore the feasibility, safety and long-term efficacy of laparoscopic total gastrectomy combined with distal pancreaticosplenectomy for the treatment of T4b gastric cancer. Methods: A retrospective cohort study was performed. Clinical data of consecutive patients with T4b gastric cancer invading pancreatic tail undergoing laparoscopic or open total gastrectomy combined with distal pancreaticosplenectomy from January 2010 to December 2014 were analyzed retrospectively. Enrollment criteria: (1) primary gastric cancer confirmed by pathology as T4b adenocarcinoma; (2) chest+abdominal+pelvic enhanced CT indicated cancer invading pancreatic tail without distant metastasis, and R0 resection was evaluated as feasible before operation; (3) physical status was ECOG score 0 to 2, and was tolerant to operation. Patients with peritoneal implant metastasis and tumor invasion of other organs during operation, or changes in surgical methods for other reasons were excluded. All the operations were performed by the same surgical team, which had the experiences of more than 100 cases of laparoscopic and 100 cases of open radical gastrectomy with D2 lymph node dissection. The choice of surgical procedure was discussed by the surgeon and the patient, and decided according to the patient's intension. Patients were divided into the laparoscopic group and open group according to the surgical method. Intraoperative and perioperative findings were compared between the two groups. The 3-year disease-free survival rate were analyzed with Kaplan-Meier survival curve and compared by using log-rank test. Results: A total of 37 consecutive patients were enrolled, including 21 in the laparoscopic group and 16 in the open group, and no one receiving laparoscopic procedure was converted to open surgery. The baseline data of two groups were comparable (all P>0.05). Compared with the open group, the laparoscopic group had significantly longer operation time [(264.0±35.1) minutes vs. (226.6±49.9) minutes, t=2.685, P=0.011], significantly less intraoperative blood loss [(65.7±37.4) ml vs. (182.2±94.6) ml, t=-4.658, P<0.001], significantly shorter time to postoperative flatus [(2.8±0.7) days vs. (4.1±0.7) days, t=-5.776, P<0.001] and significantly shorter postoperative hospital stay [(13.3±2.8) days vs. (16.6±4.3) days, t=-2.822, P=0.008]. Morbidity of postoperative complications, including anastomotic leakage, pancreatic fistula, abdominal abscess, intraperitoneal hemorrhage and duodenal stump leakage, in two groups was similar [19.0% (4/21) vs. 4/16, P=0.705]. There were no cases of anastomotic bleeding or stenosis. The 30-day postoperative mortality was 0 in the laparoscopic group and 1/16 in the open group, respectively (P=0.432). The 3-year disease-free survival rates were 38.1% and 37.5% in the laparoscopic and open group, respectively (P=0.751). Conclusion: Laparoscopic total gastrectomy combined with distal pancreaticosplenectomy performed by experienced surgeons for T4b gastric cancer is safe and effective.

Molecular typing and identification of Bdellovibrio-and-like organisms isolated from seawater shrimp ponds and adjacent coastal waters.

AIMS: To apply and compare two PCR-based methods for typing saltwater Bdellovibrio-and-like organisms (BALOs) and to understand ecological and phylogenetic aspects of the BALOs isolated from shrimp mariculture systems. METHODS AND RESULTS: Using double-layer agar technique, the numbers of culturable BALOs that lyse Vibrio alginolyticus were found to be 10-10(3) PFU ml(-1) in the surface water samples. A total of 130 BALOs isolates were differentiated into five phylotypes by denaturing gradient gel electrophoresis targeting the 16S rDNA V3 region and four phylotypes by amplified rDNA restriction analysis of the Bacteriovoracaceae-specific 16S rDNA fragment respectively. Phylogenetic analysis of representative isolates showed that all of them were identified as Bacteriovorax spp., but affiliated with four different clusters in the family Bacteriovoracaceae. CONCLUSIONS: The two PCR-based methods both can be chosen to rapidly type the saltwater BALOs at cluster level. And the relatively large numbers of BALOs with various phylotypes recovered from the same habitats suggested that these predators might play important ecological role in shrimp mariculture environments. SIGNIFICANCE AND IMPACT OF THE STUDY: We proposed two effective methods to distinguish rapidly large numbers of BALOs isolates and our results would be helpful to understand the diversity and function of BALOs in mariculture environments.

Measurement of pulsatile haemodynamic forces in a model of a bifurcated stent graft for abdominal aortic aneurysm repair.

The longitudinal haemodynamic force (LF) acting on a bifurcated stent graft for abdominal aortic aneurysm repair has been estimated previously using a simple one-dimensional analytical model based on the momentum equation which assumes steady flow of an inviscid fluid. Using an instrumented stent-graft model an experimental technique was developed to measure the LF under pulsatile flow conditions. The physical stent-graft model, with main trunk diameter of 30mm and limb diameters of 12 mm, was fabricated from aluminium. Strain gauges were bonded on to the main trunk to determine the longitudinal strain which is related to the LF. After calibration, the model was placed in a pulsatile flow system with 40 per cent aqueous glycerol solution as the circulating fluid. The LF was determined using a Wheatstone bridge signal-conditioning circuit. The signals were averaged over 590 cardiac cycles and saved to a personal computer for subsequent processing. The LF was strongly dependent on the pressure but less so on the flowrate. The measured forces were higher than those predicted by the simplified mathematical model by about 6-18 per cent during the cardiac cycle. The excess measured forces are due to the viscous drag and the effect of pulsatile flow. The peak measured LF in this model of 30 mm diameter may exceed the fixation force of some current clinical endovascular stent grafts.

Glycosides of cistanche improve learning and memory in the rat model of vascular dementia.

OBJECTIVE: Glycosides of cistanche (GC) is extracted from Xin Jiang Cistanche, which is widely used as a Chinese herb. This study aims to evaluate the effects of GC on vascular dementia (VD). MATERIALS AND METHODS: The VD model was established by the ligature of the bilateral common carotid artery in adult Wistar rats, who received daily i. p. administration of saline, GC (10 mg/kg body weight/d, i.p.) or oxiracetam (450 mg/kg body weight/d, i.p) for 14 days. Morris Water Maze test valued cognitive performance of the rats. The hippocampus was dissected and subjected to proteomics and immunohistochemical analysis. RESULTS: The GC group showed significantly lower escape latency than the VD group at four and five days after surgery. They showed no significant difference when compared with sham-operated group and the oxiracetam control group. In the hippocampus, the 21 protein spots in the GC group showed different expression levels compared with the VD group. This included the four proteins that showed a significant difference: three upregulated proteins thioredoxin-like protein 1, dual specificity mitogen-activated protein kinase kinase 1 and dihydropyrimidinase-related protein 2 (CRMP-2), and one down-regulated protein glutathione synthetase. Immunohistochemistry analysis showed that P-tau protein level was significantly higher in the VD model group than the sham-operated group (p < 0.05). After GC treatment, P-tau protein level in VD model rats showed a significant decrease compared with VD group treated with saline (p < 0.05). CONCLUSIONS: The GC plays a critical role in protecting the hippocampal neurons in the VD, by decreasing P-tau phosphorylation and increasing the CRMP-2 expression level. Pharmacological manipulation of GC offers a new opportunity for VD treatment.

[Rapid destroying of mRNAs in disaggregated Dictyostelium cells and effect of antibiotics].

Nogalamycin can inhibit RNA synthesis in both aggregated and disaggregated Dictyostelium discoideum cells. Cycloheximide and puromycin are effective in blocking translation. The late mRNAs, specific for the multicellular stage, degrade rapidly in disaggregated Dictyostelium cells. Nogalamycin or actinomycin D and daunomycin protect some of these mRNAs. Cycloheximide stabilizes all of the mRNAs, Puromycin, however, does not protect the mRNAs from destroying. These results imply that the rapid degradation of the late mRNAs during cell differentiating does not depend on nuclease or protein synthesis de novo.

A new nonadride derivative from mangrove fungus (strain No. k38).

A new nonadride derivative, ( - )-1-hydroxybyssochlamic acid (1) and the known ( - )-byssochlamic acid (2) were isolated from mangrove fungus (strain No. k38) collected from the South China Sea coast. The structure and relative configuration of 1 were elucidated by spectral data and X-ray diffraction analysis. Primary bioassays showed that 2 had medium cytotoxic activity against HEp-2 and HepG2 Cells, and 1 exhibited weak activity.

Human mesenchymal stromal cells ameliorate the phenotype of SOD1-G93A ALS mice.

BACKGROUND: Amyotrophic lateral sclerosis (ALS) is a progressive, lethal, neurodegenerative disease, currently without any effective therapy. Multiple advantages make mesenchymal stromal cells (MSC) a good candidate for cellular therapy in many intractable diseases such as stroke and brain injury. Until now, no irrefutable evidence exists regarding the outcome of MSC transplantation in the mouse model of ALS. The present study was designed to investigate the therapeutic potential of human MSC (hMSC) in the mouse model of ALS (SOD1-G93A mice). METHODS: hMSC were isolated from iliac crest aspirates from healthy donors and kept in cell cultures. hMSC of the fifth passage were delivered intravenously into irradiated pre-symptomatic SOD1-G93A mice. Therapeutic effects were analyzed by survival analysis, rotarod test, motor neuron count in spinal cord and electrophysiology. The engraftment and in vivo differentiation of hMSC were examined in the brain and spinal cord of hMSC-transplanted mice. RESULTS: After intravenous injection into irradiated pre-symptomatic SOD1-G93A mice, hMSC survived more than 20 weeks in recipient mice, migrated into the parenchyma of brain and spinal cord and showed neuroglia differentiation. Moreover, hMSC-transplanted mice showed significantly delayed disease onset (14 days), increased lifespan (18 days) and delayed disease progression compared with untreated mice. DISCUSSION: Our data document the positive effects of hMSC transplantation in the mouse model of ALS. It may signify the potential use of hMSC in treatment of ALS.

Identification of the critical role of Tyr-194 in the catalytic activity of a novel N-acyl-homoserine lactonase from marine Bacillus cereus strain Y2.

Enzymatic disruption of quorum-sensing (QS) pathways in pathogenic organisms is a promising anti-infection therapeutic strategy. AHL-lactonase, a potent tool for biocontrol, can hydrolyze QS signal molecule N-acyl-homoserine lactones (AHLs) into inactive products, thereby blocking the QS systems. A marine bacterial isolate Y2, identified as a Bacillus cereus subsp., was found capable of inactivating AHLs. The aiiA gene encoding the AHL-degrading enzyme from bacterial strain Y2 was cloned and expressed in Escherichia coli. The 28-kDa recombinant Y2-AiiA protein was purified and showed strong AHL-degrading activity. Sequence comparisons of Y2-aiiA with known AHL-lactonases revealed high identities in the deduced amino-acid sequences. Functional determination of a potential catalytic residue Tyr-194 of AHL-lactonases was performed by site-directed mutagenesis. As judged by AHL-degrading bioassay, substitution of Tyr-194 with Ala resulted in a dramatic decrease of activity compared with wild-type (WT) recombinant Y2-AiiA, although the expression level of the mutated Y2-AiiA protein was equivalent to that of WT Y2-AiiA. These results suggested that the conserved residue Tyr-194 is critical for catalytic function of the novel AHL-lactonase.

Open method versus capsulorrhaphy without drainage in the treatment of children with hepatic hydatid disease.

A recognized major postoperative complication of hepatic hydatid disease is accumulated hydrops leading to secondary infection in the residual cavity of the excised cyst. This study examined two methods of treating cystic hepatic hydatid disease in 43 children. A group of 22 children were treated by a new method of open drainage, 21 underwent capsulorrhaphy without drainage. There was no mortality or morbidity. The open method was associated with more rapid resolution of hydrops and quicker shrinkage of residual cavities, with efficacy over a median follow-up of 42 months.