[Exploration on laparoscopic hepatectomy on central liver tumor: a report of 40 cases].

Objective: To assess the safety and feasibility of the application of the laparoscopic modality in the perioperative treatment of central liver tumors. Methods: Collecting all the clinical information of a total of 40 patients with central liver tumors who received laparoscopic resection treatment carried out at Department of Hepatological Surgery of People's Hospital of Hunan Provincial from January 2016 to December 2018 to take a retrospective review. There were 19 males and 21 females.The age was (59.5±14.5) years (range: 15 to 71 years) . There were 26 cases of primary hepatic carcinoma (24 cases of hepatocellular carcinoma, 2 cases of cholangiocellular carcinoma) , 8 cases of hepatic cavernous hemangioma, 1 case of metastatic hepatic carcinoma, 5 cases of hepatocellular adenoma. The maximum diameter of tumors were (6.2±2.9) cm (range: 2 to 13 cm) . The patient's information about hepatectomy methods, blocking mode and time of blood flow, operation time, intraoperative blood loss, intraoperative blood transfusion rate, post-operative hospitalization time, perioperative reoperation and postoperative complications were collected. Results: A total of 40 patients all were treated with laparoscopic surgery. The surgical procedure was as follows: 2 patients received the right hepatic lobectomy (Ⅴ, Ⅵ, Ⅶ and Ⅷ segments) , 2 patients received the left hepatic lobectomy (Ⅱ, III and Ⅳ segments) , 13 patients received mesohepatectomy (Ⅳ, Ⅰ and Ⅷ segments) , 2 patients received left hepatic trisegmentectomy (Ⅱ, Ⅲ, Ⅳ and Ⅷ segments) , 2 patients received right hepatic trisegmentectomy (Ⅳ, Ⅴ, Ⅵ, Ⅶ and Ⅷ segments) , 7 patients received Ⅷ segmentectomy, 1 patient received Ⅳ segmentectomy, 3 patients received Ⅴ and Ⅷ segmentectomy, 5 patients received hepatic caudate lobe resection (Ⅰ, Ⅸ segments) , and 3 patients received local tumors resection.Pathological results: there were 26 cases of primary hepatic carcinoma (24 cases of hepatocellular carcinoma, 2 cases of cholangiocellular carcinoma) , 8 cases of hepatic cavernous hemangioma, 1 case of metastatic hepatic carcinoma, 5 cases of hepatocellular adenoma; the pathological reports of all malignant tumor cases all showed negative incisal edge. The operative time was (333±30) minutes (range: 280 to 380 minutes) ; the intraoperative hepatic portal occlusion period was (58±13) minutes (range: 30 to 90 minutes) ; the intraoperative hemorrhage was (173±129) ml (range: 20 to 600 ml) ; the intraoperative blood transfusion rate was 2.5% (1/40) ; the postoperative incidence of bile leakage was 2.5% (1/40) , the hospital discharge of 1 patient with bile leakage was approved after conservative treatments like T pipe decompression and adequate drainage; there was 1 case of abdominal infection and 1 case of pulmonary infection, both of which were discharged from the hospital with conservative treatments; there were no other serious postoperative complications. The postoperative hospital stay was (10.7±2.7) days (range: 6 to 16 days) ; there were no perioperative mortality and reoperation cases. Conclusion: In the centers with abundant laparoscopic hepatectomy experiences, the laparoscopic resection is proved to be safe and feasible in the perioperative treatments of central liver tumors by the highly selective cases, the adequate preoperative assessment and reasonable surgical techniques and approach.

[Amputation of a limb secondary to snakebite in a child]. / Amputation de membre secondaire à une morsure de vipère.

Amputation of a limb is a serious consequence of snakebite poisoning. It is caused by the toxicity of the venom and often the use of a tourniquet in some patients, which can lead to limb ischemia. Management of the victim aims to ensure survival and preserve the function of the bitten limb. Antivenom immunotherapy is the only specific treatment for ophidian envenomation. It is indicated in cases of general symptoms and signs related to the bite, but also if local damage could lead to the loss of limb function. The authors report on a case of snakebite that led to amputation at the thigh.

Scaffolding protein Homer1a protects against NMDA-induced neuronal injury.

Excessive N-methyl-D-aspartate receptor (NMDAR) activation and the resulting activation of neuronal nitric oxide synthase (nNOS) cause neuronal injury. Homer1b/c facilitates NMDAR-PSD95-nNOS complex interactions, and Homer1a is a negative competitor of Homer1b/c. We report that Homer1a was both upregulated by and protected against NMDA-induced neuronal injury in vitro and in vivo. The neuroprotective activity of Homer1a was associated with NMDA-induced Ca2+ influx, oxidative stress and the resultant downstream signaling activation. Additionally, we found that Homer1a functionally regulated NMDAR channel properties in neurons, but did not regulate recombinant NR1/NR2B receptors in HEK293 cells. Furthermore, we found that Homer1a detached the physical links among NR2B, PSD95 and nNOS and reduced the membrane distribution of NMDAR. NMDA-induced neuronal injury was more severe in Homer1a homozygous knockout mice (KO, Homer1a-/-) when compared with NMDA-induced neuronal injury in wild-type mice (WT, Homer1a+/+). Additionally, Homer1a overexpression in the cortex of Homer1a-/- mice alleviated NMDA-induced neuronal injury. These findings suggest that Homer1a may be a key neuroprotective endogenous molecule that protects against NMDA-induced neuronal injury by disassembling NR2B-PSD95-nNOS complexes and reducing the membrane distribution of NMDARs.

Activity of p44/42 MAP kinase in the caudal subnucleus of trigeminal spinal nucleus is increased following perioral noxious stimulation in the mouse.

The extracellular signal-regulated protein kinase-1 and -2 (ERK1 and ERK2), also referred to as the p44/42 mitogen-activated protein kinase (p44/42 MAP kinase), plays an essential role in neuronal signal transduction, but its function involved in nociceptive response has not been deeply studied yet. Here we report immunohistochemical evidence that p44/42 MAPK might be critical in nociceptive response. We found that after formalin was injected into the perioral skin of the upper lip of mice, the number of activated p44/42 MAPK-like immunoreactive neurons was significantly increased in the laminae I and II of the caudal subnucleus of the trigeminal spinal nucleus (Sp5C). The positive neurons and fibers were mostly concentrated in the middle portion of Sp5C dorsoventrally, where the afferent fibers innervating the skin of the upper lip are terminated. The reactive products were localized in perikarya, dendrites, nuclei, and diffusely in the neuropil. The present result suggests that p44/42 MAPK may be important in the transmission and modulation of noxious information in Sp5C.

Fusion Expression and Purification of OSBP PH Domain and Preliminary Analysis of Its Second Structure.

Oxysterol binding protein (OSBP) is a regulator of oxysteroid metabolism. To investigate the function and the structure-function relationship of OSBP, the recombinant vector OSBP PH-pRSET-A was transformed into E.coli JM109(DE3), and the strain highly expressing soluble 6His-OSBP PH domain in minimal medium were obtained. The fusion protein was purified by Ni(2 )-NTA agarose beads. The secondary structure of the purified 6His-OSBP PH domain fusion protein was analysed by circular dichronism. The results indicated the PH domain was composed of alpha-helix 7.2%, beta-pleated sheets 71.1% and radom coil 21.7%.

Expression, Purification and Functional Identification of Extracellular Part of Discoidin Domain Receptor 2.

Discoidin domain receptor 2 (DDR2) is a new type of receptor tyrosine kinases, and was thought to be involved in the metastasis of some tumors. Its ligand is fibrillar collagen. The activation of DDR2 induced by collagen mediates the over-expression of matrix metalloproteinase 1 (MMP-1) in cells. A specific inhibitor of DDR2 was necessary for the study of DDR2 function. Theoretically, a soluble receptor could possibly be used as specific inhibitor for the native receptor on cell membrane. In this report, a fragment (DB) of extracellular part of DDR2 was cloned and expressed for the use as potential inhibitor. This DB fragment corresponded to the polypeptide from the 23rd amino acid residue to the 293rd amino acid residue of DDR2. The fragment was amplified by RT-PCR from human lung cancer tissue, and the product was cloned into pMD18-T vector. After identification by sequence analysis, the fragment was sub-cloned into pGEX-4T-1 vector. Fusion protein of GST-DB was expressed in JM109 E.coli cells as expected and the soluble part accounted for about 13% of the total fusion protein. The soluble fusion protein was then purified with glutathione affinity resin, and GST-DB with purity of 86.1% was obtained. Competitive combination inhibitory test showed that the purified GST-DB inhibited the interaction between collagen II and DDR2 on the surface of RA synovial fibroblasts. Zymography analysis showed that the level of MMP-1 of both NIH 3T3 cell and RA synovial fibroblasts with collagen II-stimulation decreased after adding GST-DB fusion protein. The results indicated that the fusion protein GST-DB could inhibit the function of DDR2 on cells, and DDR2 might mediate collagen II-induced over-expression of MMP-1 in these cells.

Independent maternal origin of Chinese swamp buffalo (Bubalus bubalis).

To obtain more knowledge on the origin and genetic diversity of the swamp buffalo (Bubalus bubalis) in China, the complete mitochondrial D-loop sequences of 119 samples representing seven native types were compared. Two mitochondrial DNA (mtDNA) lineages (lineages A and B) were determined for the Chinese swamp buffalo. Examination of the diversity patterns suggest that lineage A has undergone a population expansion event. Divergence of lineages A and B was estimated at 18,000 years ago. Combined analyses of mtDNA sequences from Chinese, Indian, Brazilian/Italian and Southeast Asian/Australian buffalo samples showed independent domestication events in the swamp buffalo from China and the river buffalo from the India subcontinent. The spread of swamp and river buffalo from China and India respectively to mainland Southeast Asia suggests that Southeast Asia is a hybrid zone for buffalo. Our data support the hypothesis of the evolution of domesticated swamp and river buffalo from ancestral swamp-like animals. These ancestral animals were extensively distributed across mainland Asia and most likely are represented today by the wild Asian buffalo (Bubalus arnee).

An hepatitis B virus surface antigen specific single chain of variable fragment derived from a natural immune antigen binding fragment phage display library is specifically internalized by HepG2.2.15 cells.

Hepatitis B virus surface antigen (HBsAg), a specific antigen on the membrane of hepatitis B virus (HBV)-infected cells, provides a perfect target for therapeutic drugs. In order to mediate successful targeted delivery of these therapies, it is essential to have antibodies that recognize HBsAg with high specificity and affinity. In this report, we constructed a natural immune antigen binding fragments (Fab) antibody phage display library against HBsAg and after three rounds of panning, five Fab fragments with significant HBsAg binding ability were selected and analysed. DNA sequencing revealed that all the light chains had the same sequence, while all the Fd genes exhibited different sequences. For further application, all of the Fab antibodies were reconstructed into single chain antibodies (scFvs) and expressed in Escherichia coli BL21 cells. Indirect enzyme-linked immunosorbent assay analysis demonstrated that all five scFvs maintained a high affinity for HBsAg and could bind HBsAg on the membrane of HBV-infected cells. Indirect fluorescent staining analysis revealed that one of the scFvs (scFv15) could be internalized into HBsAg-positive HepG2.2.15 cells through clathrin-mediated endocytosis pathway. The internalizing scFv15 antibody would have great potential for the targeted delivery of therapeutics to HBV-infected cells.

Selective proapoptotic activity of a secreted recombinant antibody/AIF fusion protein in carcinomas overexpressing HER2.

Apoptosis-inducing factor (AIF) represents a caspase-independent apoptotic pathway in the cell, and a mitochondrial localization sequence-truncated AIF (AIFDelta1-120) can be relocated from the cytoplasm to the nucleus and exhibit a constitutive proapoptotic activity. Here, we generated a chimeric immuno-AIF protein, which comprised an HER2 antibody, a Pseudomonas exotoxin translocation domain and AIFDelta1-120. Human Jurkat cells transfected with the immuno-AIF gene could express and secrete the chimeric protein, which selectively recognized HER2-overexpressing tumor cells and was endocytosed. Subsequent cleavage of truncated AIF from immuno-AIF and its release from the internalized vesicles resulted in apoptosis of tumor cells. Intramuscular injection of the immuno-AIF gene caused significant suppression of tumors and substantially prolonged mice survival in an HER2-overexpressing xenograft tumor model. Our study demonstrates the feasibility of the immuno-AIF gene as a novel approach to treating cancers that overexpress HER2.

Signal transduction pathways involved in low intensity He-Ne laser-induced respiratory burst in bovine neutrophils: a potential mechanism of low intensity laser biostimulation.

BACKGROUND AND OBJECTIVE: Low intensity He-Ne laser irradiation has been reported to induce respiratory burst of neutrophils for a long time, but the mechanism remains obscure. We speculated that it is mediated by some signal transduction pathways. STUDY DESIGN/MATERIALS AND METHODS: The protein tyrosine kinases (PTKs) inhibitor, genistein, the phospholipase C (PLC) inhibitor, U-73122, and the protein kinase C (PKC) inhibitor, calphostin C, were used to probe signal transduction pathways of respiratory burst of bovine neutrophils which were induced by He-Ne laser at a dose of 300 J/m(2), respectively. RESULTS: The inhibitor of PTKs can completely inhibit the He-Ne laser-induced respiratory burst of neutrophils. PLC and PKC inhibitors can obviously reduce it, but not fully inhibit it. CONCLUSION: These results suggest that PTKs play a key role in the He-Ne laser-induced respiratory burst of neutrophils and [PTK-PLC-PKC-NADPH oxidase] signal transduction pathways may be involved in this process.

[Human angiogenin: expression, purification, biological assay].

Angiogenin cDNA was obtained by RT-PCR, and cloned into the fusion expression vector pRSETB. The recombinant Angiogenin protein was fused with His6 at its N-terminal and expressed as inclusion body. The expression level was about 10% of the total bacteria protein. After dissolved in 8 mol/L urea, the recombinant protein was purified by Ni2(+)-NTA chelating resin, according to the high affinity of His6 with Ni2+. The biological assay indicated that purified rhANG could induced the new blood vessel formation of CAM and degraded tRNA in vitro.