Gallstone ileus is a rare mechanical ileus,which was caused by discharge of giant gall bladder stone to the intestine.Understanding the causes of ileus is the key factor for treatment,and surgical treatment is the treatment of choice.An old patient with gallstone ileus was admitted to the Second Affiliated Hospital of Soochow University in April 2013.Preoperative X ray detection and computed tomography showed gallbladder wall thickening,formation of a sinus tract between the gall bladder and the duodenum,and intestinal ileus in the left iliac region (the diameter of the stone was about 4 cm).The patient received medical treatment for 3 days and then exploratory laparotomy + lithotomy.Gall bladder stones were not detected during the operation,so the gall bladder was preserved.The patient was followed up till December 2013,the sinus tract was disappeared under B sonography,and the cholecystitis was cured.

Objective To investigate the feasibility of amplitude of intraoperative nerve action potentials (NAP) for early quantitative diagnosis of peripheral nerve injury. Methods The sciatic nerve injury model were established in 16 rabbits. Intraoperative NAP were recorded after 4 weeks. According to amplitude of NAP, the injuried nerve were divided into 3 groups: NAP ＜ 100 μV in A group, 100 μV ≤NAP ＜ 500 μV in B group, NAP ≥ 500 μV in C group. Nerve specimen 1cm distal to injuried point were resected that received glycine silver stain and image analysis including number, diameter and cross section area of regenerative axons. Footprint parameter and ulcer area were measured and contrasted between each two groups. Results The number, diameter and cross section area of A group regenerative axons have significant difference with B and C group, no significant difference between B and C group; Footprint parameter and ulcer area have significant difference in each two groups. Conclusion Amplitude of intraoperative NAP can be a quantitative criteria to diagnose the degree of peripheral nerve injury that provides experiment evidence for guide intraoperative decision-making in clinical practice.

Objective To establish an efficient baculovirus-insect cell expression system for the production of human immunodeficiency virus-1 ( HIV-1 ) envelope glycoprotein gp120 and to evaluate the physiochemical properties, antigenicity and immunogenicity of the recombinant protein. Methods The gene encoding HIV-1 NL4-3 gp120 was cloned into the downstream of pH promoter of the baculovirus transfer vec-tor pAcgp67B to construct the recombinant transfer vector pAc-gp120. Expression of the protein of interest was induced in baculovirus-infected High FiveTM insect cells. ELISA, analytical ultracentrifugation and size-exclusion chromatography were carried out to characterize physicochemical properties of the expressed gp120 protein. Immunogenicity of the recombinant gp120 protein was analyzed by HIV neutralization assay after im-munizing BALB/c mice with it. Results The recombinant HIV-1 gp120 protein was successfully obtained from the established insect cell expression system with a purity of more than 90％ and a mean yield of 13 mg/L in four batches. That recombinant HIV-1 gp120 protein was characterized by homogeneity in solution and possessed a good immunoreactivity to neutralizing antibodies and antisera against HIV. Immunogenicity analysis in BALB/c mice demonstrated that the recombinant gp120 protein could induce effective immune re-sponses against HIV-1 NL4-3. Conclusion A simple and scalable approach to obtain homogeneous and im-munogenic HIV-1 gp120 antigen is successfully established, which will promote further investigation of HIV vaccine candidates.

Objective To evaluate the predictive performance of hematoma density heterogeneity scale (HDHS) to hematoma expansion (HE) in patients with spontaneous intracerebral hemorrhage (sICH).Methods Sixty-two patients with sICH,received therapy in our hospital form November 2011 to December 2014,were enrolled.HDHS scores were assessed on non-contrast CT (NCCT) image source,and HE progresses were assessed based on the follow up of NCCT at 24 h.Binary Logistic-regression analysis was used to determine independent risk factors of HE.Receiver operating characteristic (ROC) curve was adopted to identify the predictive performance of HDHS for HE.Results There were 37％ patients (23/62) in the cohort underwent HE.Binary Logistic-regression analysis showed that HDHS scores were independently associated with HE (OR=2.451,95％ CI:1.345-4.468,P=0.003).ROC curve analysis showed that the area under curve (AUC) was 0.745 (95％CI:0.625-0.865,P=0.001),the diagnostic critical value was 3,the sensitivity was 87％ and the specificity was 59％.Conclusion HDHS is an efficient tool that independently predict early HE in patients with sICH,and provides reliable evidence for prognosis and treatment.

We constructed the CAP2NC prokaryotic expression vector of HIV-1 NL4-3 strain and obtained relatively pure CAP2NC protein by optimizing its purification conditions to explore its in vitro self-assembly conditions. Primers were designed according to the CAP2NC DNA sequence of HIV-1 NL4-3 strain. The target gene was amplified by PCR and cloned into prokaryotic expression vector pTO-T7. Then the recombinant strain was transformed into Escherichia coli BL21 (DE3). IPTG induced protein expression, then the protein was purified by hydrophobic chromatography. SDS-PAGE and Western blotting were performed to analyze the target protein, and the biological activity of the antigen was identified through ELISA. The self-assembly of CAP2NC protein was analyzed by transmission electron microscopy and gel filtration chromatography. The protein had good reaction with the specific antibodies of p24 and formed different structures in various conditions. When 10% yeast RNA was added to the protein complex, the recombinant protein only formed into a tubular structure, which was similar to the self-assembled structure of the HIV-1 virus capsid. The results showed that the HIV-1 CAP2NC protein had in vitro self-assembly activity, and the RNA affected the structure of CAP2NC protein assembly. The protein can be used as a simple and effective molecular model to study its structure, and then it can provide a reference for the study of HIV immature virus particles.

Disturbance of macrophage-associated lipid metabolism plays a key role in atherosclerosis. Crosstalk between autophagy deficiency and inflammation response in foam cells (FCs) through epigenetic regulation is still poorly understood. Here, we demonstrate that in macrophages, oxidized low-density lipoprotein (ox-LDL) leads to abnormal crosstalk between autophagy and inflammation, thereby causing aberrant lipid metabolism mediated through a dysfunctional transcription factor EB (TFEB)-P300-bromodomain-containing protein 4 (BRD4) axis. ox-LDL led to macrophage autophagy deficiency along with TFEB cytoplasmic accumulation and increased reactive oxygen species generation. This activated P300 promoted BRD4 binding on the promoter regions of inflammatory genes, consequently contributing to inflammation with atherogenesis. Particularly, ox-LDL activated BRD4-dependent super-enhancer associated with liquid-liquid phase separation (LLPS) on the regulatory regions of inflammatory genes. Curcumin (Cur) prominently restored FCs autophagy by promoting TFEB nuclear translocation, optimizing lipid catabolism, and reducing inflammation. The consequences of P300 and BRD4 on super-enhancer formation and inflammatory response in FCs could be prevented by Cur. Furthermore, the anti-atherogenesis effect of Cur was inhibited by macrophage-specific Brd4 overexpression or Tfeb knock-out in Apoe knock-out mice via bone marrow transplantation. The findings identify a novel TFEB-P300-BRD4 axis and establish a new epigenetic paradigm by which Cur regulates autophagy, inhibits inflammation, and decreases lipid content.

Inflammation-driven endothelial dysfunction is the major initiating factor in atherosclerosis, while the underlying mechanism remains elusive. Here, we report that the non-canonical stimulator of interferon genes (STING)-PKR-like ER kinase (PERK) pathway was significantly activated in both human and mice atherosclerotic arteries. Typically, STING activation leads to the activation of interferon regulatory factor 3 (IRF3) and nuclear factor-kappa B (NF-κB)/p65, thereby facilitating IFN signals and inflammation. In contrast, our study reveals the activated non-canonical STING-PERK pathway increases scaffold protein bromodomain protein 4 (BRD4) expression, which encourages the formation of super-enhancers on the proximal promoter regions of the proinflammatory cytokines, thereby enabling the transactivation of these cytokines by integrating activated IRF3 and NF-κB via a condensation process. Endothelium-specific STING and BRD4 deficiency significantly decreased the plaque area and inflammation. Mechanistically, this pathway is triggered by leaked mitochondrial DNA (mtDNA) via mitochondrial permeability transition pore (mPTP), formed by voltage-dependent anion channel 1 (VDAC1) oligomer interaction with oxidized mtDNA upon cholesterol oxidation stimulation. Especially, compared to macrophages, endothelial STING activation plays a more pronounced role in atherosclerosis. We propose a non-canonical STING-PERK pathway-dependent epigenetic paradigm in atherosclerosis that integrates IRF3, NF-κB and BRD4 in inflammatory responses, which provides emerging therapeutic modalities for vascular endothelial dysfunction.