Objective To study the molecular pathway of osteoblastic differentiation induced by substance P (SP), a neurotransmitter. Methods Mesenchymal stem cells were isolated and cultured, and treated with SP or its receptor (NK1) antagonist to induce osteoblastic cell differentiation, respectively. Alkaline phosphatase activity was determined; Osterix gene expression was detected by RT-PCR after 1-2 weeks for three times. The data of each culture condition were analyzed using SPSS12.0 statistical software to determine whether the differences between conditions were significant. Results After 4-5 days' culture, bone marrow stromal cells became spindle-shaped, triangular or polygonic. They covered the plate surface, formed extensive cell sheets in each group after 11-12 days of culture, and then induced differentiation to osteoblast. SP up-regulated the important transcription factor Osterix gene expression significantly (P<0.05). Conclusion The up-regulation of Osterix gene expression by SP may stimulate osteoblastic cell differentiation. SP's regulation depends on its receptor NK1.

Objective To evaluate the tissue response to the covered metallic stent loaded with radioactive 125I seeds in normal rabbit esophagus.Methods Twelve rabbits were randomlv assigncd into 2 groups.A covered metallic stent loaded with three 125I seeds set in sheaths Was implanted into the esophagus of the rabbit from experimental group(22.2 MBq for each seed,n=6).The rabbits of control group were implanted with stent without 125I (n=6).Two rabbits from each group underwent esophagus X-ray examination to detect migration of the stent and the 125I seeds at 2,4 and 8 weeks after stent implantation,respectively.And the animals were then sacrificed to observe the changes in esophagus.Results No 125I seed migration Was found during and after the implantation of the stent.No esophageal perforation was observed.Two weeks after stent implantation,such mild lesions were observed in the middle of the esophagus in experimental group as slight epithelial hyperplasia and submucosal inflammation.At 4th week after stentimplantation,granulation and fibrosis were observed,which became more obvious at 8th week after the procedure.The injury of esophageal tissue opposite to the 125I seed Was significantly milder than that of the tissue exposed to 125I seed.In control group,esophagus adjacent to the middle of the stent WaS similar to normal tissue,with slight epithelial hyperplasia.However,both ends of the stent were covered with marked hyperplasia epithelium in both groups,and severe granulation and fibrosis could be observed.Conclusion The main pathological changes of esophagus wall adjacent to the 125I seed are granulation and fibrosis.No bleeding or perforation occurs.

To investigate the effects of anti-sense peptide nucleic acids (PNAs) targeting Ki-67 gene on modulation of the proliferation and apoptosis of human renal carcinoma cell lines, human renal carcinoma cell line 786-0 cells were treated with anti-sense PNAs at different concentrations (1.0 micromol/L, 2.0 micromol/L, 10.0 micromol/L). The Ki-67 expression of 786-0 cells was detected by immunohistochemical technique and Western blot method respectively. The proliferation of 786-0 cells was studied by cell growth curves and 3H-thymidine incorporation. The apoptosis of 786-0 cells was detected by TUNEL assay. The control groups were treated with anti-sense oligonucleotide (ASODNs) targeting Ki-67 gene. Our results showed that the Ki-67 expression of 786-0 cells treated with anti-sense PNAs (16.9+/-0.7) was significantly inhibited as compared with that of the control groups (28.6+/-0.4) (P<0.01). The Ki-67 protein rate of 786-0 cells treated with anti-sense PNAs (42.1 +/-2.2) was significantly reduced when compared with that of the control groups (83.6+/- 1.4) (P<0.01). Proliferation of 786-0 cells treated with anti-sense PNAs (20.7+/- 1.5) was significantly inhibited as compared with that of the control groups (58.6+/- 1.4) (P<0.01). The apoptosis rate of 786-0 cells treated with anti-sense PNAs (28.7+/- 2.3) was significantly increased higher compared with that of the control groups (13.8 +/- 1.0) (P<0.01). From these finds we are led to conclude that anti-sense PNAs targeting Ki-67 gene have stronger effects on the inhibition of the proliferation and induction of apoptosis of human renal carcinoma cells than ASODNs targeting Ki-67 gene. The strategies using anti-sense PNAs targeting Ki-67 gene may be a promising approach for the treatment of renal cell carcinoma.

ObjectiveTo investigate the effect of interstitial brachytherapy with 125Ⅰon human esophageal carcinoma implanted in nude mice. MethodsAnimal model simulating human esophageal carcinoma was established by subcutaneous implantation of cultured Eca-109 cell lines into nude mice.The mice were randomly divided into 3 groups,namely control group (saline plus empty seed),125Ⅰ seed group (22.2 MBq×1 seed),and DDP group (cisplatin at the dose of 1 mg/kg),to receive corresponding treatment.The growth rate and the pathological changes of esophageal carcinoma were observed.ResultsThe animals were sacrificed 16 days after irradiation.The average tumor weight in control group,125Ⅰ seed group,and DDP group were (0.20±0.06) g, (0.12±0.03) g and (0.12±0.05 ) g,respectively (P<0.05).Pathological findings included degeneration and necrosis of the tumor cells.Compared to the control group,the necrosis areas in 125Ⅰ seed group and DDP group were significantly larger than those in control (P<0.05).Conclusion125Ⅰ seed brachytherapy in esophageal carcinoma could cause degeneration and necrosis of the tumor cells and had inhibitory effect on tumor growth.

Early intervention contributes to improving patient experience and doctor-patient relationship in the case of non-psychiatric outpatients with psychological problems.The authors studied the psychological assessment and hierarchical management for non-psychiatric inpatients at a general hospital. Measures taken include establishing multi-disciplinary and inter-department teams, building an intra-hospital joint-action system, and implementing the psychological assessment and hierarchical management for non-psychiatric inpatients.These efforts explored ways for a general hospital in psychological counseling, offering humanistic service, and transformation of medical pattern.

To investigate the effects of anti-sense peptide nucleic acids (PNAs) targeting Ki-67gene on modulation of the proliferation and apoptosis of human renal carcinoma cell lines, human renal carcinoma cell line 786-0 cells were treated with anti-sense PNAs at different concentrations (1.0 μmol/L, 2.0 μmol/L, 10.0 μmol/L). The Ki-67 expression of 786-0 cells was detected by immunohistochemical technique and Western blot method respectively. The proliferation of 786-0 cells was studied by cell growth curves and 3H-thymidine incorporation. The apoptosis of 786-0 cells was detected by TUNEL assay. The control groups were treated with anti-sense oligonucleotide (ASODNs)targeting Ki-67 gene. Our results showed that the Ki-67 expression of 786-0 cells treated with anti-sense PNAs (16.9±0.7) was significantly inhibited as compared with that of the control groups (28.6±0.4) (P＜0.01). The Ki-67 protein rate of 786-0 cells treated with anti-sense PNAs (42.1±2.2)was significantly reduced when compared with that of the control groups (83.6±1.4) (P＜0.01). Proliferation of 786-0 cells treated with anti-sense PNAs (20.7±1.5) was significantly inhibited as compared with that of the control groups (58.6±1.4) (P＜0.01). The apoptosis rate of 786-0 cells treated with anti-sense PNAs (28.7±2.3) was significantly increased higher compared with that of the control groups (13.8±1.0) (P＜0.01). From these finds we are led to conclude that anti-sense PNAs targeting Ki-67 gene have stronger effects on the inhibition of the proliferation and induction of apoptosis of human renal carcinoma cells than ASODNs targeting Ki-67 gene. The strategies using anti-sense PNAs targeting Ki-67 gene may be a promising approach for the treatment of renal cell carcinoma.

Cryoablation (CRA) and microwave ablation (MWA) are two main local treatments for hepatocellular carcinoma (HCC). However, which one is more curative and suitable for combining with immunotherapy is still controversial. Herein, CRA induced higher tumoral PD-L1 expression and more T cells infiltration, but less PD-L1^highCD11b⁺ myeloid cells infiltration than MWA in HCC. Furthermore, CRA had better curative effect than MWA for anti-PD-L1 combination therapy in mouse models. Mechanistically, anti-PD-L1 antibody facilitated infiltration of CD8⁺ T cells by enhancing the secretion of CXCL9 from cDC1 cells after CRA therapy. On the other hand, anti-PD-L1 antibody promoted the infiltration of NK cells to eliminate PD-L1^highCD11b⁺ myeloid cells by antibody-dependent cell-mediated cytotoxicity (ADCC) effect after CRA therapy. Both aspects relieved the immunosuppressive microenvironment after CRA therapy. Notably, the wild-type PD-L1 Avelumab (Bavencio), compared to the mutant PD-L1 atezolizumab (Tecentriq), was better at inducing the ADCC effect to target PD-L1^highCD11b⁺ myeloid cells. Collectively, our study uncovered the novel insights that CRA showed superior curative effect than MWA in combining with anti-PD-L1 antibody by strengthening CTL/NK cell immune responses, which provided a strong rationale for combining CRA and PD-L1 blockade in the clinical treatment for HCC.