Anlotinib has strong antiangiogenic effects and leads to vessel normalization.However,the"window period"characteristic in regulating vessel normalization by anlotinib cannot fully explain the long-term survival benefits achieved through combining it with other drugs.In this study,through RNA sequencing(RNA-seq)and label-free quantitative proteomics analysis,we discovered that anlotinib regulated the expression of components of the extracellular matrix(ECM),leading to a significant reduction in ECM stiffness.Our bioinformatic analysis revealed a potential positive relationship between the ECM pathway and gefitinib resistance,poor treatment outcomes for programmed death 1(PD-1)targeting,and unfavourable prognosis following chemotherapy in lung cancer patients.We administered anlotinib in combination with these antitumour drugs and visualized their distribution using fluorescent labelling in various tumour types.Notably,our results demonstrated that anlotinib prolonged the retention time and distribution of antitumour drugs at the tumour site.Moreover,the combination therapy induced notable loosening of the tumour tissue structure.This reduction was associated with decreased interstitial fluid pressure and tumour solid pressure.Additionally,we observed that anlotinib effectively suppressed the Ras homologue family member A(RhoA)/Rho-associated protein kinase(ROCK)signalling pathway.These findings suggest that,in addition to its antiangiogenic and vessel normalization effects,anlotinib can increase the distribution and retention of antitumour drugs in tumours by modulating ECM expression and physical properties through the RhoA/ROCK signalling pathway.These valuable insights contribute to the development of combination therapies aimed at improving tumour targeting in cancer treatment.

Animals ; Antibodies, Neutralizing/biosynthesis* ; Antibodies, Viral/biosynthesis* ; Body Weight/immunology* ; COVID-19/virology* ; Disease Models, Animal ; Disease Progression ; Humans ; Immunohistochemistry ; Lung/virology* ; Male ; Mesocricetus ; Nasal Cavity/virology* ; RNA, Viral/immunology* ; SARS-CoV-2/pathogenicity* ; Severity of Illness Index ; Viral Load

Aim To evaluate the potency of anti-D. acutus venom IgY neutralizing the main activities of D. acutus venom.Methods After mixing the different a-mounts of IgY with snake venom and incubating togeth-er,the main activities of snake venom were assayed by biochemical methods.Results The in vitro assays in-dicated that anti-D.acutus venom IgY obviously neu-tralized the activities of PLA2 ,5′-nucleotidase,hyalu-ronidase,metalloprotease and serine proteinase (fi-brinogenase)in D.acutus venom.Mouse experiments showed that the ED50 value of IgY for mouse was 1 131.09 μg.Conclusion Anti-D.acutus venom IgY antibodies have good effects in neutralizing D.acutus venom without the toxicities themselves.

OBJECTIVETo investigate the safety, effectiveness and long-term results of Jinling procedure.

METHODSA total of 1 100 patients with refractory constipation were admitted to the Institute of General Surgery between February 2000 and December 2013 and received Jinling procedure. There were 217 male and 883 female patients with an average age of (45±15) years and an average duration of (275±159) months. The Jinling procedure modifies the classic procedure of subtotal colectomy with colorectal anastomosis by adding a new side-to-side cecorectal anastomosis. The general clinical data, postoperative complications, gastrointestinal quality of life index (GIQLI), Wexner constipation score, satisfaction rate, body composition, nutritional status, homeostasis, anal manometry and defecography during follow-up were collected. Statistical analysis was performed by t-test analysis, χ(2) test or Fisher's exact probability.

RESULTSThe postoperative follow-up rate were 96.73%, 94.36%, 93.00% and 92.55% at 1, 3, 6, and 12 months. The satisfaction rate were 62.50%, 72.45%, 93.16% and 94.70% respectively during 1, 3, 6 and 12 months follow-up. The postoperative 1 month, GIQLI was lower than that of preoperative (P=0.038), but significantly better at postoperative 3 month than the preoperative (P=0.022), and gradually improve after then. Wexner score was significantly lower at postoperative 1 month than the preoperative (P=0.018), and with the gradually reduce. Compared to the preoperative baseline, all body composition and serum protein parameters decreased significantly at the one-month follow-up and then increased gradually over the remaining follow-up period. Weight and prealbumin were significantly higher than the preoperative at postoperative 12 month. Jinling procedure had no effect on electrolytes. All patients were detected in defecography before surgery, but these abnormal indicators were improved after surgery. Anal function, coordinated movement anorectal function, anorectal reflex and rectal sensation were injured at early postoperative, but as time goes on, these indicators gradually returned to preoperative levels, some indicators better than the preoperative level. Postoperative surgical site infection, anastomotic bleeding , anastomotic leakage, urinary retention, sexual dysfunction, recurrent constipation, bowel obstruction, anastomotic stricture and mortality rates were 4.36%, 5.45%, 6.00%, 6.91%, 0.64%, 0.45%, 8.82%, 4.00% and 0.27% respectively.

CONCLUSIONSJinling procedure is a safe and effective surgical procedure for refractory constipation. It can improve defecation function and quality of life significantly, and has a high follow-up satisfaction rate.

Adult ; Anastomosis, Surgical ; Colectomy ; Constipation ; surgery ; Defecation ; Defecography ; Female ; Follow-Up Studies ; Humans ; Intestinal Obstruction ; Male ; Middle Aged ; Postoperative Period ; Quality of Life ; Rectum ; Treatment Outcome

OBJECTIVETo investigate the safety, efficacy and long-term outcomes of Jinling procedure in the treatment of adult Hirschsprung disease.

METHODSClinical and follow-up data of 125 patients with adult Hirschsprung disease undergoing Jinling procedure at the Department of General Surgery between January 2000 and January 2013 were summarized. All the patients were diagnosed by CT, barium enema, anorectal pressure detection and pathology examination. Abdominal symptoms, gastrointestinal quality of life index(GIQLI, the lower score, the worse quality of life), Wexner constipation score (higher score indicated worse symptom), defecography (evaluation included rectocele, mucosal prolapse, intramucosal intussusception, perineal prolapse) and other operative complications were compared before and after operation.

RESULTSAmong 125 patients, 69 were male and 56 were female with median age of (41.2±15.5) (18 to 75) years. The follow-up rates were 94.4%(118/125), 92.0%(115/125), 89.6%(112/125) and 88.0%(110/125) at postoperative months 1, 3, 6, and 12. Incidences of abdominal distension and abdominal pain were 100% and 82.4%(103/125) before operation, and were 7.3%(8/110) and 20.9%(23/110) at 12 months after surgery. Wexner score was significantly lower at postoperative months 1(8.7±2.9), 3 (7.2±2.8), 6(6.7±2.2) and 12(6.3±1.7) than that before operation (21.4±7.2) (P<0.01). GIQLI score was 51.6±11.9 before operation, though it decreased at 1 month (47.3±5.5)(P<0.05) after surgery, but increased significantly at postoperative months 3, 6, 12(68.9±8.0, 96.5±8.2, 103.2±8.6)(P<0.01). Abnormal rate of defecography was 70.4%(81/115), 48.2%(54/112) and 27.3%(30/110) at postoperative months 3, 6, 12, which was significantly lower than 91.2%(114/125) before operation (P<0.01). Morbidity of postoperative complication was 29.6%(37/125), including 5 cases of surgical site infection (4.0%), 2 of anastomotic bleeding (1.6%), 8 of anastomotic leakage (6.4%, one died of severe abdominal infection), 4 of urinary retention (3.2%), 3 of recurrent constipation (2.4%, without megacolon relapse), 11 of bowel obstruction (8.8%), 2 of anastomotic stricture(1.6%) and 2 of refractory staphylococcus aureus enteritis (1.6%, diagnosed by stool smear and culture, and both died finally).

CONCLUSIONJinling procedure is a safe and effective surgical procedure for adult Hirschsprung's disease.

Adolescent ; Adult ; Aged ; Anastomosis, Surgical ; Colectomy ; Constipation ; Defecography ; Digestive System Surgical Procedures ; Female ; Hirschsprung Disease ; surgery ; Humans ; Intestinal Obstruction ; Intussusception ; Male ; Middle Aged ; Perineum ; Postoperative Complications ; Postoperative Period ; Quality of Life ; Rectocele ; Staphylococcus aureus ; Treatment Outcome ; Young Adult

The aim of this research is to investigate the preparation method of valve membrane in transcatheter bio- prosthetic valve, and to study the effect of chemical modification and cutting technology to tensile property and suture force property of valve membrane. We carried out a series of processes to perform the tests, such as firstly to test the crosslinking degree of valve membrane using ninhydrin method, then to test the tensile property and suture force property by using Instron's biomechanicAl testing equipment, and then to observe the collagen fiber orientation in valve membrane using Instron's biomechanical testing equipment and using field emission scanning electron microscopy. The study indicated that after the chemical modification, the crosslinking degree, tensile strength and suture force strength increasing rate of valve membrane were 93.78% ± 3. 2%, (8.24 ± 0.79) MPa, 102%, respectively. The valve membrane had a better biomechanical property and would be expected to become valve membrane in transcatheter bioprosthesis valve.
Bioprosthesis ; Collagen ; chemistry ; Heart Valve Prosthesis ; Microscopy, Electron, Scanning ; Tensile Strength

BACKGROUNDMitochondrial dysfunction has been reported to be one of the contributing factors of sepsis-associated encephalopathy (SAE). Mitochondrial biogenesis controls mitochondrial homeostasis and responds to changes in cellular energy demand. In addition, it is enhanced or decreased due to mitochondrial dysfunction during SAE. The aim of this study was to explore the changes of mitochondrial biogenesis of astrocytes under septic conditions.

METHODSLipopolysaccharide (LPS; 50 ng/ml) and interferon-γ (IFN-γ; 200 U/ml) were incubated with astrocytes to model the effects of a septic insult on astrocytes in vitro. The mitochondrial ultrastructure and volume density were evaluated by transmission electron microscopy. Intracellular adenosine triphosphate (ATP) levels were detected by the firefly luciferase system. The expression of protein markers of mitochondrial biogenesis and the binding ability of mitochondrial transcription factor A (TFAM) were determined by western blot and electrophoretic mobility shift assays, respectively. The mitochondrial DNA (mtDNA) content was detected by real-time polymerase chain reaction.

RESULTSThe number of mildly damaged mitochondria was found to be significantly greater after treatment for 6 hours, as compared with at 0 hour (P < 0.05). The mitochondrial volume density was significantly elevated at 24 hours, as compared with at 0 hour (P < 0.05). The ATP levels at 6 hours, 12 hours, and 24 hours were significantly greater than those at 0 hour (P < 0.05). The protein markers of mitochondrial biogenesis were significantly increased at 6 hours and 12 hours, as compared with at 0 hour (P < 0.05). The TFAM binding activity was not significantly changed among the four time points analyzed. The mtDNA contents were significantly increased at 12 hours and 24 hours, as compared with at 0 hour (P < 0.05).

CONCLUSIONSUnder septic conditions, mitochondrial biogenesis of astrocytes increased to meet the high-energy demand and to promote mitochondrial recovery. Furthermore, the TFAM-DNA binding ability was not sensitive to sepsis-induced injury.

Animals ; Astrocytes ; drug effects ; metabolism ; Blotting, Western ; Cells, Cultured ; DNA, Mitochondrial ; genetics ; Electrophoretic Mobility Shift Assay ; Interferon-gamma ; pharmacology ; Lipopolysaccharides ; pharmacology ; Microscopy, Electron, Transmission ; Mitochondrial Turnover ; drug effects ; physiology ; Nitric Oxide ; metabolism ; Rats ; Reactive Nitrogen Species ; metabolism ; Sepsis ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective To investigate effect of the venom peptide liquor on adjunctive arthritis (AA) in mice .Methods The male Kunming mice were randomly divided into 6 groups ,named as the control group ,model group ,positive control group(1 mg/kg dex-amethason) ,wine group(10 mL/kg) ,low dosage of venom peptide liquor group(8 .3 mL/kg) and high dosage of venom peptide liq-uor group(33 .2 mL/kg);the right toes ,ankle diameter and whole body weight were measured at 7-day intervals ;the spleen index , serum level of circulating immune complexes(CIC) were determined after the thirty-fourth day when the mice was put to death .Re-sults The level of CIC in AA mice decreased significantly (P<0 .05) ,the decrease of spleen index and the reduction of toe and an-kle swelling in the low dose group were significant(P<0 .01) .Conclusion The venom peptide liquor exhibited apparent inhibitory effect on AA in mice .

OBJECTIVETo clone and express VP1-VP4 genes encoding the structural proteins of Coxsackie virus A16 and analyze the antigenicity of the expressed recombinant proteins.

METHODSThe VP1-VP4 cDNAs were amplified with RT-PCR from the extracted viral RNA and cloned into pMD19-T vectors. The VP1-VP4 genes were inserted to the multi-cloning sites of the plasmid pQE30a, and the protein expressions in E. coli M15 were induced by IPTG. After purification by washing with 8 mol/L urea under denaturing condition, the recombinant proteins were identified by Western blotting and ELISA for their immunogenicity against rabbit antisera of Coxsackie virus A16 and enterovirus 71, respectively.

RESULTSThe recombinant VP1-VP4 proteins were highly expressed in E. coli M15. The purified proteins could be recognized by rabbit antiserum of Coxsackie virus A16 and showed cross reactivity with the rabbit antiserum of Enterovirus 71.

CONCLUSIONThe recombinant Coxsackie virus A16 VP1-VP4 proteins obtained possess good antigenicity.

Antigens, Viral ; genetics ; immunology ; Capsid Proteins ; classification ; genetics ; immunology ; Cloning, Molecular ; Enterovirus A, Human ; genetics ; Gene Expression ; Genetic Vectors ; Plasmids ; Recombinant Proteins ; genetics ; immunology ; Reverse Transcriptase Polymerase Chain Reaction

Objective To explore the changes of etiology and mortality of upper gastrointestinal bleeding (UGIB) in the last 20 years in Guangdong region. Methods A total of 3140 UGIB cases diagnosed in Guangdong General Hospital from January 1990 to October 2009 were analyzed with retrospective analysis. Groups were divided according to admission chronological order and age to analyze the causes of UGIB to get the trend of changes and influencing factors. Results UGIB occurred more in men than in women, the gender ratio was 2. 5∶1. There was no significant change in gender composition between the first and later 10 years. Peptic ulcer bleeding (PUB) was the main cause of UGIB in young and middle-age patients (age<60 years), especially duodenal ulcer (DU)bleeding (45. 0%) , and secondary was acute gastric mucosal lesion ( AGML) (16. 6%). While in elder patients group (age≥60 years), the main cause of UGIB was AGML (40. 5%), the frequency was significantly higher compared with the young and middle-age group (40. 5% vs 16. 6%,P<0. 01).Compared the first 10 years with the later 10 years, the occurrence rate of PUB was overall in downward trend (57. 4% vs 42. 8%, P<0.01), of which DU and compound ulcer(CU) bleeding reduced significantly (DU: 43.8% vs 27.7%, P<0. 01 and CU: 7.7% to 5.5%, P<0. 05). While the incidence of gastric ulcer (GU) (7.4% vs 8. 1% ,P>0. 05) and esophagogastric variceal bleeding (EVB) (11. 7% vs 12. 9%, P>0.05) remained stable. The occurrence of AGML bleeding significantly increased than before (32. 4% vs 18. 8%,P<0. 01), and became one of the most important causes of UGIB. Conclusions PUB is still the most important cause of UGIB in Guangdong region. AGML becomes another important cause of UGIB in elder patients, which may relate to the increasing use of non-steroidal anti-inflammatory drugs (NSAIDs), anti-platelet and antineoplastic medicine.