Hepatic Stellate Cells ; Humans ; Liver Cirrhosis

Objective To evaluate the changes in water, electrolyte and acid-base balance after seawater immersion in cases of open abdominal trauma associated with intestinal rupture, and to obtain a theoretical basis for the early treatment of open abdominal injury with intestinal rupture in naval combat. Methods A canine model of open abdominal trauma with intestinal rupture was established in 26 healthy adult dogs, and they were divided randomly into three groups. All animals were subjected to abdominal wall incision and intestinal rupture. Seawater immersion group(n=10) was immersed into artificial seawater after trauma; normal saline solution group(n=6) was immersed into normal saline solution after trauma; control group (n=10) had no immersion. The 3 groups were observed for changes in water, electrolyte and acid-base balance, and the results were analysed and compared. Results Signficant disturbance of water, electrolyte and acid-base imbalance were observed in the seawater immersion groups, but no significant changes of these parameters were seen in the control group and normal saline group. Conclusion Seawater immersion is the main factor leading to the disturbance of body metabolism after open abdominal trauma with intestinal rupture.

Objective: To express the protein of HPV18E6 based on pET-32a(+) at high level and study the expression and significance of HPV18E6 proteins in laryngeal carcinoma. Methods: The HPV18E6 gene was amplified by PCR and cloned into pET-32a(+). The amplified fragment was inserted into the plasmid pET32a (+) that was digested with BamHⅠand Hind Ⅲ. The recombinant plasmid pET32/E6 was transformed into E.coli JM109 which was selected with ampicillin. The recombinant plasmids were successfully introduced into E.coli BL21(DE 3) and were induced by IPTG. SDS-PAGE and Western blot analysis were used to detect the confusion protein. Finally, the optimization of expression conditions, such as temperature, concentration of IPTG, was studied. Results: The recombinant plasmids were identified and confirmed with enzyme digestion and sequencing. The BL21(DE3) transformed recombinant plasmid pET32/E6 had expressed HPV18E6 recombinant protein effectively. The optimum conditions of expression were 37 ℃, 1 mmol/L IPTG. Conclusion:Prokaryotic expression vector pET-32a(+)-HPV18E6 was successfully constructed. The high-level expression of HPV18E6 was achieved in E.coli BL21(DE3).

Objective To evaluate a protocol of early treatment for dogs suffering from open abdominal injury and seawater immersion. Methods A canine model of open abdominal injury was made with 30 adult health dogs. After 2 hours seawater immersion, the dogs were divided into two groups: treatment group( n =18) and control group(untreated group, n =18). The changes of survival rate, body liquid metabolism and hemodynamics were observed respectively. Results The survival rate significantly increased from 0 in control group to 94 4% in treatment group. The disorder of hemodynamics and body liquid metabolism was improved significantly, P

Objective To establish the xenotransplanted tumor model of Craniopharyngioma in chick chorioallan?toic membrane (CAM) and detect the angiogenesis ability, microvessel density (MVD) and cell proliferation of the xeno?graft. Method Craniopharyngioma tissues from surgical craniopharyngioma patients were transplanted on the CAM. An?giogenesis assay was performed and the MVD and PCNA were evaluated using immunohistochemistry following the trans?plantation. Results The tumor formation rate of adamantinomatous craniopharyngioma (ACP) and squamous papillary cra?niopharyngioma (SPCP) was 47.14% and 43.33%, respectively. There was no significant difference in tumor formation rate between ACP and SPCP(χ2=0.123,P=0.726). The CAM angiogenesis, MVD and expression of PCNA were higher in ACP than in SPCP. The expression of PCNA was positively correlated with MVD (Pearson r=0.639,P<0.001) and CAM assay score (Spearman r=0.490,P=0.001 ) in CP. Conclusion The model of human craniopharyngioma can be es?tablished in the CAM. The angiogenesis of the xenograft in the CAM can be evaluated and the craniopharyngioma xeno?graft of CAM possesses a new blood circulation and cell proliferation ability.

Prokaryotic expression vector of mouse HPV16E6 gene was constructed. A pair of primers were designed according to the digestion sites in plasmid pGEX-KG and the HPV16E6 gene sequence published by GenBank. The DNA fragment of 321bp was amplified by PCR from the HPV recombinant plasmid with HPV16E6 gene, then cloned into pGEX-KG and transformed into the host E. coli strain JM109. The fragment was conformed to the original sequence, which indicated that fusion expression vector pGEX-KG-HPV16E6 was constructed. The pGEX-KG-HPV16E6 plasmid was taken and transformed into BL21(DE3) for expression. Induced by IPTG at 37 degrees C, the expression product of HPV16E6 gene was identified by SDS-PAGE and Western blot. HPV16E6 fusion protein had been expressed successfully in the form of inclusion bodies, the molecular weight of fusion protein being 38 kD. Meanwhile, the optimum condition of HPV16E6 fusion protein expression was induced with 1.0 mmol/L IPTG for 4h. The fusion protein reacted specifically with the antibodies against HPV16E6. HPV16E6 gene was successfully expressed in E. coli, which could be used as a basis for preparing HPV16E6 vaccine in human.
Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Glutathione Transferase ; biosynthesis ; genetics ; Humans ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; immunology ; Repressor Proteins ; biosynthesis ; genetics ; Viral Vaccines ; immunology

Objective To study the risk factors of the biochemical remission of pituitary tumor patients with acromegaly treated by the endoscopic endonasal surgery. Methods A retrospective analysis of clinical data was conducted on 61 cases acromegaly patients underwent endoscopic endonasal surgery between August 2013- November 2016.Endocrinology tests were performed in all patients,including the fasting/random GH(growth hormone,GH)level, Nadir GH during OGTT(oral glucose tolerance test, OGTT), and IGF-1(insulin-like growth factor-1,IGF-1). The clinical data included gender, age, preoperative GH value, preoperative IGF-1 value, tumor invasion of the inferface space of internal carotid artery(ICA),tumor surrounding angle of internal carotid artery(≥/<135°),Knosp grade, Hardy grade, and tumor volume. Univariate analysis and multivariate Logistic retrospective analysis were used to analyze the association between above-mentioned factors and the occurrence of biochemical remission. Results There were 52.5% (32/61)patients achieving biochemical remission.Univariate analysis showed significant differences between patients with and without biochemical remission in preoperative GH value, tumor surrounding angle of internal carotid artery(

Objective The aim of our study was to establish an endoscopic endonasal transsphenoidal surgical training model, and to examine its application value. Methods The endoscopic endonasal transsphenoidal surgical training models were made.After assessments,the models was used for endoscopic operation training for 8 neurosurgeons with no experiences of endoscopic operation. Results The available operating space of this model is similar to the actual operating space in endoscopic endonasal transsphenoidal surgery. The egg fixed on the model has many layers of structure, such as eggshell, shell membrane, egg white and egg yolk, which constituted a good practice object. All students indicated that they could benefit from the training using this model. The skill in grinding the eggshell [(1.07 ± 0.221)cm2/min vs.(1.45±0.27)cm2/min, P<0.001]was significantly improved and the chance for shell membrane rupture (8 vs. 2, P=0.007) was significantly reduced in the last-time practice compared with the first-time practice (P<0.001). Conclusion The endoscopic endonasal transsphenoidal surgical training model is helpful for neurosurgeons to practice the basic operation of endoscopic surgery and to improve their surgical skills,and can be used repeatedly.It can be used in basic operation training before the training using cadaver cranium.

Objective To investigate the distance change of bilateral internal carotid artery in acromegalic patients. Methods Twenty patients with acromegaly were included in this study from August 2016 to March 2018 in Department of Neurosurgery of the First Affiliated Hospital of Fujian Medical University. There were 7 males and 13 females with an average age of 40.2±12.6 years (range from 21 to 62 years old). Forty sex and age matched patients with non-functional pituitary adenoma were selected as controls. The 3D printing software was used for reconstructing the internal carotid artery. Distances of the bilateral internal carotid arteries were measured on the reconstructed images. Results The maximum distance between bilateral siphon carotid ectasias (25.1±3.9 mm vs. 23.2±2.9 mm, P=0.041) and distance between bilateral lacerum segments (26.2 ±3.1 mm vs. 23.8 ±4.1 mm, P=0.022) were significantly longer in patients with acromegaly than in those without. Although the distance between bilateral cavernous segments (20.7 ±4.3 mm vs. 22.4 ±3.1 mm, P=0.076) tended to be shorter, the difference was not statistically significant. The distance between bilateral ophthalmic segments was not significantly different between the two groups (15.7±5.1 mm vs. 16.0±2.5 mm, P=0.783). Conclusion The distance of bilateral internal carotid artery of acromegalic patients is different from that of patients with non-functional pituitary adenomas. Patients with acromegaly have significantly longer distance between bilateral siphon carotid ectasias and distance between bilateral lacerum segments compared with the patients with non- functional pituitary adenomas.

Objective:To effectively express the receptor binding domain (RBD) of SARS-CoV-2 spike protein in Pichia pastoris and to evaluate its immunogenicity. Methods:The gene encoding the RBD protein was synthesized and cloned into the pPICZαA plasmid. After linearization, the plasmid was transferred and integrated into the genome of Pichia pastoris. The expressed RBD protein in culture supernatant was analyzed by Western blot and Biolayer interferometry. After screening, a single clone expressing the RBD protein was selected. The high-level expression of RBD protein was achieved by optimizing the fermentation process, including the salt concentration adjusting of the medium and induction condition optimization (pH, temperature and duration). The immunogenicity of the expressed RBD protein was evaluated in a mouse model. Results:A single clone with a high expression level of RBD protein was obtained and named RBD-X33. The expression level of RBD protein in the fermentation supernatant reached up to 240 mg/L after optimization of the induction condition (HBSM medium, pH=6.5±0.3, 22℃ and 120 h). In the mouse experiment, the recombinant RBD protein was formulated with Alum+ CpG dual adjuvant and injected into mice. The binding IgG antibody levels were up to 2.7×10 6 tested by ELISA and the neutralizing antibody levels were up to 726.8 tested by live virus neutralizing antibody assay (prototype). Conclusions:The RBD protein could be efficiently expressed in Pichia pastoris and induce stronger immune response in animals. This study suggested that the recombinant SARS-CoV-2 RBD protein expressed in Pichia pastoris could serve as a candidate antigen in the development of SARS-CoV-2 vaccine.