[Comparison of efficacy between laparoscopic and open proximal gastrectomy with double-tract reconstruction for Siewert type â ¡ and â ¢ adenocarcinoma of the esophagogastric junction].

Objective: To compare the efficacy between laparoscopic and open proximal gastrectomy with double-tract reconstruction for Siewert type II and III adenocarcinoma of the esophagogastric junction (AEG). Methods: A retrospective cohort study was conducted. Inclusion criteria: (1) 18 to 80 years old; (2) Siewert II and III AEG was confirmed by preoperative gastroscopy and biopsy, which could not be resected by endoscopy; patients undergoing radical proximal gastrectomy with double-tract reconstruction; (3) contrast-enhanced abdominal CT staging was cT1-2N0M0; (4) Eastern Cooperative Oncology Group (ECOG) physical status score <2 points, American Association of Anesthesiologists (ASA) grade 1 to 2; (5) patients agreed to perform proximal gastrectomy and signed an informed consent. Those who had undergone neoadjuvant radiochemotherapy, suffered from serious mental diseases and had incomplete data were excluded. According to the above criteria, clinical data of 84 consecutive patients with Siewert II and III AEG undergoing surgery at General Surgery Department of The Affiliated Tumor Hospital of Zhengzhou University from October 2010 to December 2018 were collected and analyzed. Of 84 patients, 61 underwent open proximal gastrectomy with double-tract reconstruction (OPG group), while 23 underwent laparoscopic proximal gastrectomy with double-tract reconstruction (LPG group). The perioperative complications and postoperative reflux esophagitis of two groups were compared. A P-value of <0.05 was considered to be statistically significant. Results: Among 84 cases, 74 were male and 10 were female. There were 43 cases of Siewert type II and 41 cases of Siewert type III. There were no significant differences in age, gender, body mass index, comorbidities, Siewert type, and tumor staging between the two groups (all P>0.05). As compared to the OPG group, the LPG group had longer operation duration [(223±21) minutes vs. (161±14) minutes, t=15.352, P<0.001], less intraoperative blood loss [195 (150, 215) ml vs. 208 (192, 230) ml, Z=2.143, P=0.032], and shorter time to flatus [(2.8±0.7) days vs. (3.3±0.9) days, t=2.477, P=0.015]. There were no significant differences in the number of harvested lymph nodes, time to the first meal and postoperative hospital stay between the two groups (all P>0.05). Postoperative complications developed in 2 cases (8.7%, 1 case each for anastomotic leakage and intestinal obstruction) in the LPG group and 5 cases (8.2%, 1 case each for anastomotic leakage, anastomotic bleeding, and anastomotic stenosis, 2 cases of incision infection) in the OPG group (χ(2)=5.603, P=0.231). The median follow-up was 41.2 (12.8-110.5) months. One patient (1.6%,1/61) had obvious reflux symptoms in the OPG group, compared with none in the LPG group (χ(2)=0.644, P=0.422). Esophagitis occurred in 1 case (4.8%, 1/21) in LPG group, compared with 4 patients (7.1%, 4/56) in the OPG group, without significant difference between the two groups (χ(2)=0.505, P=0.477). Conclusion: Laparoscopic proximal gastrectomy with double-tract reconstruction is safe and feasible without increasing the risk of postoperative complication and reflux esophagitis.

Effects of 12 weeks of aerobic exercise on body composition and vascular compliance in obese boys.

AIM: This study tested the hypothesis that 12 weeks of air board exercise would enhance cardiorespiratory fitness and vascular compliance and reduce % body fat in obese Korean boys. METHODS: Twenty-two obese boys (>30% body fat) were studied. They were divided into 2 groups- an aerobic exercise group (N.=12), which trained 3 days/week, 50 min/day for 12 weeks, and a control group (N.=10). Control subjects only performed activities involved in their physical education classes. Body composition, cardiovascular fitness (20 m multistage endurance test performance) and vascular compliance were assessed before and after the completion of exercise training. RESULTS: The % changes in body fat (-4.6±0.9 vs. -1.5±1.0%), fat mass (-5.4±1.5 vs. -0.1±1.6%) and performance on the cardiovascular fitness test (14.3±2.5 vs. 3.7±1.6%) were greater in the exercise group than in the controls Compared to controls, % increases in vascular compliance were greater in the arms and legs of the exercise group (left arm: 2.8±0.5 vs. 2.0±2.9%; left leg: 2.6±1.2 vs. -0.5±2.0%; right arm: 2.9±0.9 vs. 0.3±2.9%; right leg: 4.8±1.8 vs. 1.5±2.0%). CONCLUSION: Results suggest that exercise training can reduce % body fat and enhance vascular compliance in obese male adolescents; changes that may reduce the risk for later development of cardiovascular disease.

Identification of four highly conserved genes between breakpoint hotspots BP1 and BP2 of the Prader-Willi/Angelman syndromes deletion region that have undergone evolutionary transposition mediated by flanking duplicons.

Prader-Willi and Angelman syndromes (PWS and AS) typically result from an approximately 4-Mb deletion of human chromosome 15q11-q13, with clustered breakpoints (BP) at either of two proximal sites (BP1 and BP2) and one distal site (BP3). HERC2 and other duplicons map to these BP regions, with the 2-Mb PWS/AS imprinted domain just distal of BP2. Previously, the presence of genes and their imprinted status have not been examined between BP1 and BP2. Here, we identify two known (CYFIP1 and GCP5) and two novel (NIPA1 and NIPA2) genes in this region in human and their orthologs in mouse chromosome 7C. These genes are expressed from a broad range of tissues and are nonimprinted, as they are expressed in cells derived from normal individuals, patients with PWS or AS, and the corresponding mouse models. However, replication-timing studies in the mouse reveal that they are located in a genomic domain showing asynchronous replication, a feature typically ascribed to monoallelically expressed loci. The novel genes NIPA1 and NIPA2 each encode putative polypeptides with nine transmembrane domains, suggesting function as receptors or as transporters. Phylogenetic analyses show that NIPA1 and NIPA2 are highly conserved in vertebrate species, with ancestral members in invertebrates and plants. Intriguingly, evolutionary studies show conservation of the four-gene cassette between BP1 and BP2 in human, including NIPA1/2, CYFIP1, and GCP5, and proximity to the Herc2 gene in both mouse and Fugu. These observations support a model in which duplications of the HERC2 gene at BP3 in primates first flanked the four-gene cassette, with subsequent transposition of these four unique genes by a HERC2 duplicon-mediated process to form the BP1-BP2 region. Duplicons therefore appear to mediate genomic fluidity in both disease and evolutionary processes.

Retrotransposed genes such as Frat3 in the mouse Chromosome 7C Prader-Willi syndrome region acquire the imprinted status of their insertion site.

Prader-Willi syndrome (PWS) results from loss of function of a 1.0- to 1.5-Mb domain of imprinted, paternally expressed genes in human Chromosome (Chr) 15q11-q13. The loss of imprinted gene expression in the homologous region in mouse Chr 7C leads to a similar neonatal PWS phenotype. Several protein-coding genes in the human PWS region are intronless, possibly arising by retrotransposition. Here we present evidence for continued acquisition of genes by the mouse PWS region during evolution. Bioinformatic analyses identified a BAC containing four genes, Mkrn3, Magel2, Ndn, Frat3, and the Atp5l-ps1 pseudogene, the latter two genes derived from recent L1-mediated retrotransposition. Analyses of eight overlapping BACs indicate that these genes are clustered within 120 kb in two inbred strains, in the order tel-Atp5l-ps1-Frat3-Mkrn3-Magel2-Ndn-cen. Imprinting analyses show that Frat3 is differentially methylated and expressed solely from the paternal allele in a transgenic mouse model of Angelman syndrome, with no expression from the maternal allele in a mouse model of PWS. Loss of Frat3 expression may, therefore, contribute to the phenotype of mouse models of PWS. The identification of five intronless genes in a small genomic interval suggests that this region is prone to retroposition in germ cells or their zygotic and embryonic cell precursors, and that it allows the subsequent functional expression of these foreign sequences. The recent evolutionary acquisition of genes that adopt the same imprint as older, flanking genes indicates that the newly acquired genes become 'innocent bystanders' of a primary epigenetic signal causing imprinting in the PWS domain.

[Further report on application of DNA probe in diagnosis of vivax malaria infection].

Southern blotting with a labeled and linearized pUC 19 DNA containing a specific fragment of 0.24 kb DNA of Plasmodium vivax asexual blood stages (kindly offered by Dr. C. Kidson) was used for further identification of blood samples showing positive reaction by dot-blot hybridization. The results showed that those with positive reaction from patients with P. vivax, with P. falciparum or with fever but with negative microscopic findings were also positive by Southern blotting. It was confirmed that some of those positive with P. falciparum were likely to infect P. vivax at the same time. So did a part of those with fever but negative in the blood films (Figs. 1,2).

Expression of a partial synthetic human TNF cDNA in E. coli.

A recombinant human tumour necrosis factor (rhTNF) cDNA was constructed. The TNF gene was isolated from a human genomic gene library. There are four exons in the TNF gene. The fourth exon codes for 140 amino acids of the TNF matured protein which is composed of 157 amino acids. A major portion of the fourth exon was isolated and then ligated to a synthesized DNA fragment coding for the remaining amino acids. The partial synthetic hTNF (rhTNF) cDNA thus generated was subcloned into a vector and successfully expressed in E. coli. 5-1 fermentator was used to produce rhTNF. About 20 g (wet weight) of bacterial pellet per liter medium and 10(6)-10(7) units of cytotoxicity to L929 cells per milliliter medium were obtained. rhTNF was purified by HPLC and dried with a freeze dryer. rhTNF with a purity of about 95% in the form of white powder was obtained. The sequence of ten amino acids at the amino terminus of the rhTNF was determined. The result showed that it was identical with that of the natural human TNF.

[The selective isolation of cosmid clones by homologous recombination in Escherichia coli--a cosmid clone containing t complex linkage DNA sequence of mouse was isolated].

A procedure for the selective isolation of specific cosmid clones by homologous recombination between cosmid clones of genomic library and a probe DNA sequence cloned in a plasmid in vivo has been developed. The cosmid library was constructed in a rec- host cell strain and packaged into phage particles in vivo. The rec+ host cells containing a DNA sequence used as selection probe cloned in the pUC plasmid were infected by packaged cosmid phage particles. There is no homology between cosmid and the plasmid vectors. After a period of 1-3 hr. for the recombination to take place, the probe plasmids were integrated into cosmid, in which the DNA sequence are homologous with the probe, by homologous recombination. The cosmids are then packaged in vivo and transferred into a rec- cell strain. The specific cosmid clones were selected by double antibiotic resistance carried by both vectors. The probe plasmid can be excised by lambda excision enzyme by using superinfection with red+ phage. After packaging in vivo, these cosmid revertants can be identified on Xgal plate. A cosmid clone containing the t complex linkage DNA sequence of mouse was selected by using the procedure above with a probe derived from microdissected metaphase chromosome.

Molecular cloning and restriction mapping of human lymphotoxin gene.

In order to clone the human lymphotoxin (HuLT) gene, we practiced a concise and time-saving method: homologous recombination in vivo (1). By using the mouse lymphotoxin (MuLT) cDNA (1.3 kb) as a probe, we isolated the HuLT gene from a human genomic library which was constructed with cosmid pcos2EMBL as a vector. After linearization, the recombinant cosmid was partially digested with BamHI, EcoRI, PstI, and PvuII respectively, and either cos end was labelled by hybridization with radioactive oligos complementary to the cohesive end sequence (2). The physical map of HuLT gene was made by this method.

Analysis of cosmids using linearization by phage lambda terminase.

A group of cosmid clones was isolated from the region of the mouse t complex and analysed by a rapid restriction mapping protocol based on linearization of circular cosmid DNA in vitro. A plasmid capable of producing high levels of phage lambda terminase was constructed and procedures for in vitro cleavage of cosmid DNAs were optimised. After linearization, the cosmids were partially digested with restriction enzymes, and either cos end was labelled by hybridization with radioactive oligos complementary to the cohesive end sequence, a step which we have described previously for clones in phage lambda (Rackwitz et al., 1984). High-resolution restriction maps derived by this method were used to identify and align the cosmids, to localise the position of repetitive sequences, and to interpret the results of electron microscopy heteroduplex experiments.