Objective To study the expression of substance P(SP) and substance P receptor(SPR) during the development of mice brains. Methods The expression of SP and SPR during the development of mice brains from embryonic day(E) 11 to postnatal day(P) 0 days was analyzed by immunohistochemical method. Results The expression of SP began at E11 and gradually increased until birth. The expression of SPR began at E11 and maintained stable expression until birth. SP mostly expressed at striatum and SPR mostly expressed at medullary raphe.Conclusion The expression of SP and SPR during the embryo brain stage may indicate that SP could be an important factor involved in the early organization and maturation of neuron.

The coronavirus disease 2019 (COVID-19) is an acute infectious disease caused by the new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, which has led to serious worldwide economic burden. Due to the continuous emergence of variants, vaccines and monoclonal antibodies are only partial effective against infections caused by distinct strains of SARS-CoV-2. Therefore, it is still of great importance to call for the development of broad-spectrum and effective small molecule drugs to combat both current and future outbreaks triggered by SARS-CoV-2. Cathepsin L (CatL) cleaves the spike glycoprotein (S) of SARS-CoV-2, playing an indispensable role in enhancing virus entry into host cells. Therefore CatL is one of the ideal targets for the development of pan-coronavirus inhibitor-based drugs. In this study, a CatL enzyme inhibitor screening model was established based on fluorescein labeled substrate. Two CatL inhibitors IMB 6290 and IMB 8014 with low cytotoxicity were obtained through high-throughput screening, the half inhibition concentrations (IC₅₀) of which were 11.53 ± 0.68 and 1.56 ± 1.10 μmol·L^-1, respectively. SDS-PAGE and cell-cell fusion experiments confirmed that the compounds inhibited the hydrolysis of S protein by CatL in a concentration-dependent manner. Surface plasmon resonance (SPR) detection showed that both compounds exhibited moderate binding affinity with CatL. Molecular docking revealed the binding mode between the compound and the CatL active pocket. The pseudovirus experiment further confirmed the inhibitory effects of IMB 8014 on the S protein mediated entry process. In vitro pharmacokinetic evaluation indicated that the compounds had relatively good drug-likeness properties. Our research suggested that these two compounds have the potential to be further developed as antiviral drugs for COVID-19 treatment.

It is the objective of this paper to study pharmacokinetics-pharmacodynamics (PK-PD) characteristics of ginsenoside Rg1 and Rb1 on the effect of inducing nitric oxide (NO) release after intravenous administration of Shengmai injection to rats with myocardial ischemia. The model of myocardial ischemia rats was produced by subcutaneous injection of isoproterenol. The serum samples were collected at different time points after intravenous administration of Shengmai injection to rats with the dose of 10.8 mL x kg(-1). The concentrations of ginsenoside Rg1 and Rb1 in serum were determined, and then the concentration-time curves were drawn. Pharmacokinetic parameters of ginsenoside Rg1 and Rb1 were calculated after the construction of pharmacokinetic models. Meanwhile, NO2- and NO3-, the metabolites of NO, in serum were determined, and then the effect-time curve was drawn. The combined PK-PD model was established based on the theory of effect compartment by Sheiner et al. Then pharmacodynamic parameters were calculated. The results indicated that the pharmacokinetics of ginsenoside Rg1 and Rb1 conformed to a two-compartment model. Ginsenoside Rg1 and Rb1 exhibited quick and slow elimination in rats respectively. The effect of Shengmai injection on inducing NO release did not relate directly with and lagged behind the concentrations of ginsenoside Rg1 and Rb1 in serum. The effect exhibited good correlation with ginsenoside Rg1 and Rb1 levels in effect compartment. The relationship between effect and serum concentration fits Sigmoid-E(max) model. This study successfully established the combined PK-PD model of ginsenoside Rg1 and Rb1 after intravenous administration of Shengmai injection to rats. The model can efficiently predict the concentration and effect of Shengmai injection in vivo.
Administration, Intravenous ; Animals ; Ginsenosides ; administration & dosage ; pharmacokinetics ; Humans ; Male ; Myocardial Ischemia ; drug therapy ; metabolism ; Nitric Oxide ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo explore the therapeutic effects of internal fixation with Herbert screws for the treatment of Pipkin type I and type II femoral head fractures.

METHODSFrom January 2008 to December 2012, 23 patients with Pipkin type I and type II femoral head fractures were treated with open reduction and internal fixation by Herbert screws through Kocher-Langenbeck approach. Twenty-three patients (aged 18 to 52 years with a mean of 35.5 years, including 18 males and 5 females patients, 8 left hips and 15 right hips) with femoral head fractures and posterior hip dislocation. The fracture was classified according to Pipkin classification based on the radiographic findings, 5 patients had type I and 18 had type II fractures. The duration time from admission to the operation ranged from 6 to 72 h (averaged 32 h). The clinical and radiographic outcomes of the patients were measured using Thompson-Epstein scoring scale. The Harris hip score(HHS) was used to evaluate and compare hip functions at the latest follow-up between affected and healthy sides.

RESULTSAll the patients were followed up, and the duration ranged from 20 to 48 months (averaged 30 months). According to Thompson-Epstein system, 12 patients got an excellent result, 6 good, 4 fair and 1 poor. The average HHS at the finial follow-up was 87.80 ± 8.46 (ranged from 66 to 95), which is similar to that in the healthy side 90.10 ± 6.35 (ranged from 72 to 98) (t = 1.044, P = 0.302). The complications such as deep infection, and deep vein thrombosis were not found. At the 3rd year during follow-up,4 patients had avascular necrosis of femoral head. At the 1st year of follow-up, 1 patient had hip pain after walking,screws loosening and shift after trauma,and serious complications of traumatic arthritis. All the 5 patients were treated with total hip arthroplasties.

CONCLUSIONThe treatment of internal fixation with Herbert screws through the Kocher-Langenbeck approach is effective for Pipkin type I and type II femoral head fracture. The method is reliable and valuable for recommendation. However, such fracture may have avascular necrosis of femoral head and complication of traumatic arthritis, which should be observed carefully in clinic with preparation of the prevention and treatment measures.

Adolescent ; Adult ; Bone Screws ; Female ; Femoral Neck Fractures ; surgery ; Fracture Fixation, Internal ; adverse effects ; methods ; Humans ; Male ; Middle Aged

OBJECTIVETo investigate the effects of antibiotic-PMMA (polymethyl-methacrylate) beads combined with external fixator in treatment of infected fracture nonunion.

METHODSTwenty-two cases of infected fracture-nonunions were reviewed involving 20 male and 2 female with an average age of 34.68 years (ranging 21 to 74 years). The data consisted of 9 cases of tibial fractures, 2 distal fractures of the femur, 6 femoral shaft fractures, 3 intertrochanteric fracture of the femur and 2 humeral shaft fractures. The procedure included thorough debridement to wipe out dead bone and granulation tissue, then antibiotic-PMMA bead chains imbedded into the dead space. One week later, secondary debridement was performed, antibiotic-PMMA bead chains were changed according to result of bacterial culture and susceptibility test, and fractures were stabilized with external fixator. Three months after debridement, antibiotic-PMMA bead chains were taken out and bone graft with autogenous iliac cancellous bone chips was performed.

RESULTSThe mean follow-up period was 19.98 months (ranging 15 to 28 months). Infection was controlled in 20 cases. One tibial fracture and 1 intertrochanteric fracture of the femur needed repeated debridement 2 and 3 months after bone grafting respectively,because of infection recurrence and sinus formation. All 22 cases achieved bony union averaged 15.09 weeks after bone grafting with a range of 8 to 24 weeks.

CONCLUSIONThorough debridement, imbedding antibiotic-PMMA bead chains combined with external fixator and staged bone grafting has proven to be effective and simple for treatment of infected fracture nonunion. The antibiotic bead delivers high tissue levels,obliterates dead space, aids bone repair.

Adult ; Aged ; Anti-Bacterial Agents ; therapeutic use ; Bone Diseases, Infectious ; drug therapy ; microbiology ; surgery ; Bone Transplantation ; External Fixators ; Female ; Follow-Up Studies ; Fractures, Bone ; complications ; drug therapy ; surgery ; Fractures, Ununited ; complications ; drug therapy ; surgery ; Humans ; Male ; Middle Aged ; Polymethyl Methacrylate ; chemistry ; Young Adult

Objective:To investigate the safety and prognostic value of neoadjuvant chemotherapy and surgery for advanced gastric cancer patients with para-aortic lymph node metastasis.Methods:Clinicopathological data of 25 patients admitted to the Department of Gastrointestinal Surgery, Fujian Cancer Hospital from Jan 2015 to Jun 2017 were retrospectively analyzed. All patients were treated with SOX chemotherapy for 3 cycles. D 2 + paraaortic lymphadenectomy was performed in patients with stable disease (SD) . After operation, SOX regimen was used for 5 cycles of chemotherapy. Results:After 3 cycles of neoadjuvant chemotherapy, there were 2 cases with progressive disease, 6 cases of SD and 17 cases of partial remission. There was no treatment-related death. Twenty-three patients underwent surgery, including 19(76%) patients of R 0 resection. Tirty-four out of 128 para aortic lymph nodes were metastatic. Postoperative complications occurred in 5(22%) patients, with no mortality . The median progression free survival time and median overall survival time were 20 and 29 months respectively. The 1, 3-year overall survival rates were 80% and 48%, and the 1-year and 3-year progression free survival rates were 72% and 38%, respectively. For those with para-aortic lymph node metastasis the 1-year and 3-year OS rate were 70% and 17%, respectively. Multivariate analysis showed that the efficacy of neoadjuvant chemotherapy was an independent prognostic factor. Conclusion:Neoadjuvant chemotherapy is among others an independent prognostic factor affecting the post-op survival of advanced gastric carcinoma with para-aortic lymph node metastasis.

OBJECTIVETo compare therapeutic effects between antegrade intramedullary nailing and retrograde intramedullary nailing for the treatment of humeral shaft fractures.

METHODSFrom March 1999 to October 2006, 105 patients with humeral shaft fractures were treated with locked intramedullary nail and were adequately followed up. There were 82 antegrade nailing and 23 retrograde nailing. The follow-up parameters included operation time, blood loss,fracture healing rate, healing time, complications, Constant-Murley shoulder score and Mayo elbow performance score.

RESULTSThe mean follow-up period was 31.2 months. Antegrade intramedullary nailing had significantly less blood loss than that in retrograde intramedullary nailing (P=0.002). The differences in operation time, complications, healing time and bone healing rate between he two groups had no statistical significance. Complications in the antegrade intramedullary nail group included 4 patients with nonunions, 1 patient with radial nerve palsy, and 8 patients with shoulder pains and decrement in shoulder range of motion. Complications in the retrograde intramedullary nail group included 1 patient with radial nerve palsy and 3 patients with iatrogenic fractures. For shoulder joints,the difference in the average Constant-Murley shoulder score between the two groups was statistically significant (P=0.04). For elbow joints, the average postoperative Mayo elbow performance score between these two approaches did not differ significantly.

CONCLUSIONBoth the antegrade intramedullary nailing and the retrograde intramedullary nailing are good alternatives for the treatment of humeral shaft fractures. Because of higher incidence of iatrogenic fractures, the insertion point of retrograde intramedullary nailing should be carefully prepared. With antegrade insertion, it important to bury the humeral nail below the rotator cuff to prevent the subacromial impingement, and the rotator cuff should be carefully repaired to avoid shoulder pain and improve shoulder function.

Adult ; Aged ; Case-Control Studies ; Female ; Fracture Fixation, Intramedullary ; methods ; Humans ; Humeral Fractures ; surgery ; Male ; Middle Aged

OBJECTIVETo investigate the expression of recombination activating gene-1 (RAG-1) and its localization in the mouse brain during the embryonic development.

METHODSThe brain tissues of E (embryonic day) 11, E13, E15, E17, E19, P0 (the birth day) and adult mice were taken, the total RNA of brains were extracted and the changes of RAG-1 expression were detected with the method of RT-PCR. The freeze sections of brain tissues from each group were stained with immunohistochemistry method.

RESULTThe expression of RAG-1 persisted from E11 to P0 brain and was steadily increased from E11 to E19; the results of RT-PCR were similar to that of immunohistochemistry. The positive-cells mainly appeared in the nucleus amygdalae, hypothalamus, thalamus and hippocampus at developmental stage. The expression began to appear in ventricular zone (VZ) and intermediate zone (IZ) of telecephalic vesicle, then gradually increased in subventricular zone (SVZ), corticle plate (CP) and subcorticle plate (SP).

CONCLUSIONThe expression of RAG-1 in mouse embryonic brain tissue is higher than that in the adult mouse, which may be related to the process of neuron development.

Animals ; Brain ; cytology ; embryology ; metabolism ; Female ; Gene Expression Regulation, Developmental ; Homeodomain Proteins ; genetics ; metabolism ; Immunohistochemistry ; Mice ; Mice, Inbred ICR ; Neurons ; cytology ; metabolism ; Pregnancy ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors

Tumor angiogenesis provides adequate oxygen and nutrition for tumor development and supports tumor growth and metastasis. Stromal cell derived factor 1 (SDF-1) and its receptor C-X-C motif chemokine receptor 4 (CXCR4) in pancreatic cancer microenvironment are involved in tumor growth such as promoting tumor cell proliferation, migration, and angiogenesis. In this study, anti-CXCR4 nanobody (CXCR4 Nb) and anti-programmed cell death ligand 1 (PD-L1) &CXCR4 bispecific nanobody (PX4 BsNb) were expressed in Escherichia coli system and purified by nickel column affinity chromatography. We investigated the anti-angiogenesis activity and mechanism of CXCR4 Nb by in vivo and in vitro experiments. Ethical approval was obtained for collection of human peripheral blood mononuclear cell (hPBMC) samples from the Local Ethics Committee of Shanghai Jiao Tong University. All animal experiments were approved by the Animal Ethic Committee of Shanghai Jiao Tong University. The results showed that CXCR4 Nb at 0.1 μmol·L^-1 could effectively inhibit the proliferation and migration of human umbilical vein endothelial cells (HUVEC) promoted by pancreatic stellate cells in vitro. CXCR4 Nb and PX4 BsNb at 0.3 mg·kg^-1 obviously decreased tumor angiogenesis and inhibited the tumor growth in NOD/SCID mice, the inhibitory rates were 28.8% and 36.1%, respectively. CXCR4 Nb significantly inhibited tumor growth and angiogenesis with great safety, which provides support for application of CXCR4 Nb and anti-angiogenesis therapy of pancreatic cancer.

OBJECTIVETo explore the application of serum protein pattern models in diagnosis of colorectal cancer (CRC) by proteinchip technology.

METHODSOne hundred and forty-seven serum samples (55 CRC patients and 92 healthy individuals) randomly divided into training set (n = 87, 32 CRC patients and 55 healthy individuals) and test set (n = 60), were subjected for analysis by surface enhanced laser desorption/ionization-time of flight-mass spectrometry (SELDI-TOF-MS). Four top-scored peaks in 5910, 8930, 4476 and 8817 were detected by proteinchip software version 3.0. and were trained by a multi-layer artificial neural network (ANN) with a back propagation algorithm. An artificial neural network classifier had developed for separating CRC from the healthy group. The classifier was then challenged with the test set (60 samples including 23 CRC patients and 37 healthy individuals) to determine the validity and accuracy of the classification system.

RESULTSThe artificial neural network classifier separated the CRC from the healthy samples, with sensitivity of 82.6% and specificity of 91.9%.

CONCLUSIONCombination of SELDI-TOF-MS with the artificial neural network yields significant higher sensitivity and specificity than CEA in the diagnosis of CRC, which should be further studied.

Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; analysis ; Blood Proteins ; analysis ; Colorectal Neoplasms ; diagnosis ; Female ; Humans ; Male ; Middle Aged ; Neural Networks (Computer) ; Protein Array Analysis ; Proteomics ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization