BACKGROUND:Acute toxicity in vivo experiments in previous studies has been confirmed that the median lethal dose of chitosan microcapsules is higher than 2 000 mg/kg, but the specific pathogenic mechanism is unclear.
OBJECTIVE:To explore the influence of nano chitosan on MC3T3-E1 cellgrowth as a bone substitute material, as wel as the physiological function of rats.
METHODS:MC3T3-E1 cells were respectively cultured in Dulbecco’s modified Eagle’s medium with different concentrations of chitosan nanoparticles (0, 10 mg/L, 100 mg/L, 1 g/L, 10 g/L). The absorbance values were determined. Changes in MC3T3-E1 cellmorphology were observed by scanning electron microscope after 24 hours culture. 10 g/L nano chitosan suspension was prepared using PBS. Two different doses of nano chitosan suspension (166.67 and 16.67 mg/kg body weight) with PBS were injected intraperitoneal y into Sprague-Dawley rats, three times a week, for 4 weeks. The control group was injected with equal volume of physiologic saline. Serum biochemical markers were detected to analyze the functions of liver and kidney of rats. Moreover, histopathology slices were observed to evaluate the morphological changes of tissue and inflammatory infiltration.
RESULTS AND CONCLUSION:10 mg/L, 100 mg/L, 1 g/L, 10 g/L nano chitosan suspensions were found growth inhibition in MC3T3-E1 cells as compared with the control group (P<0.05). The reunion of chitosan was observed in the cytoplasm of MC3T3-E1 cells by transmission electron microscopy. On the cellsurface, pseudopodia formed, wavy undulating membrane, nucleus degeneration, fragmentation and condensation were found. Compared with the control group, blood urea nitrogen, Na+levels were significantly increased in rats injected with nano chitosan suspension at two dosages, but the K+level in the high concentration group was decreased significantly (P<0.05). cellapoptosis was found in the liver and renal tissue in a dose-dependent manner. It suggests that apoptosis may be the possible mechanism of nano chitosan toxicity, and normal physiological function may be impacted over a certain dose.

OBJECTIVE:To study the antitumor mechanism of Ruixiang Langdu(Stellera chamaejasme L ) abstracts(SCA) METHODS:SCA-contained serum was derived from mice pre-administrated with different oral dosages of SCA and at different times after administration The effects of the serum on the proliferation of K562 leukemic cells were observed with MTT assay and clone formation RESULTS:After mixing with the SCA-contained serum derived at 1,2,4,8h after giving SCA(3,6 and 12g/kg),the rates of MTT transformation and clone formation of K562 cells were decreased significantly The SCA-contained serum 12 h after giving drug was more effective than others CONCLUSION:The SCA-contained serum inhibited the proliferation of tumor cells,which may be one of its important antitumor mechanism

Object To explore the antitumor mechanism of Stellera chamaejasme Linn. (SC). Methods SC containing-serum (SCCS) was derived from mice pretreated with different doses of SC. Cultured human leukemia HL-60 and human gastric adenocarcinoma SGC-7901 cells were used. Inhibition of proliferation was measured using MTT assay. Morphological assessment of apoptosis was performed with fluorescence microscope. DNA fragmentation was assessed by agarose gel electrophoresis and flow cytometry. Expression of bcl-2 protein was measured with immunohistochemistry. Results Exposure of exponentially growing HL-60 cells to mice serum containing 10% SC (pretreated with SC 3, 6, and 12 g/kg) for 48 h resulted in growth inhibition in a dose-dependent manner. Typical morphological changes of apoptosis and DNA fragmentation in HL-60 cells were induced. "Apobodies" in the apoptotic cells were observed, "ladder" pattern of agarose gel electrophoresis of DNA from these cells was revealed, and the percentage of apoptotic cells with fractional DNA content increased from 11.7% to 57.4%. Treatment with SC containing serum decreased the percentage of SGC-7901 cells of bcl-2 protein positive expression from 78.3% to 32.9%. Conclusion SC could induce apoptosis of HL-60 cells and decrease the expression of bcl-2 protein of gastric adenocarcinoma SGC-7901 cells.

Object To explore the antitumor mechanism of Stellera chamaejasme Linn.(SC).Methods SC containing-serum(SCCS)was derived from mice pretreated with different doses of SC.Cultured human leukemia HL-60 and human gastric adenocarcinoma SGC-7901 cells were used.Inhibition of proliferation was measured using MTT assay.Morphological assessment of apoptosis was performed with fluorescence microscope.DNA fragmentation was assessed by agarose gel electrophoresis and flow cytometry.Expression of bcl-2 protein was measured with immunohistochemistry.Results Exposure of exponentially growing HL-60 cells to mice serum containing 10% SC(pretreated with SC3,6, and 12 g/kg)for 48h resulted in growth inhibition in a dose-dependent manner.Typical morphological changes of apoptosis and DNA fragmentation in HL-60 cells were induced."Apobodies'in the apoptotic cells were observed,'ladder"pattern of agarose gel electrophoresis of DNA from 11.7% to 57.4%.Treatment with SC containing serum decreased the percentage of SGC-7901 cell of bcl-2 protein positive expression from 78.3% to 32.9%.Conclusion SC could induce apoptosis of HL-60 cells and decrease the expression of bcl-2 protein of gastric adenocarcinoma SGC-7901 cells.

Objective:To understand the characteristics of the callers with the diagnosis of mental disorders called Beijing Psychological Crisis Hotline for help,in order to provide specific psychological intervention services for them in future.Methods:From December 2002 to December 2008,24217 different callers'data collected from a computer-based operating system of the Beijing Psychological Crisis Hotline were analyzed.According to their self-reported of having the diagnoses of mental disorder or not,the callers were divided into the ones with diagnosis (n=6516)and the ones without diagnosis (n =17701).The callers'demographic characteristics,their main counseling problems and the suicide-related factors in the two groups were compared.Results:The top five main counseling problems in the group with diagnosis were mental problems,the mental illness related knowledge,the problems of family relationships,working problems and interpersonal problems.Mental problem ranked first in both groups.Compared to the group without diagnosis,the diagnosed ones were more likely to be found in such group of people as women,aged between 20 -29 and 30 -44,having 10 or more years of education and the unem-ployed.The results of the assessments of 11 suicide-related risk factors showed that callers with diagnoses had higher prevalence of most suicide-related factors than those in the other group(P <0.001 or 0.05),which included suicidal intention and behavior while calling,history of attempted suicide,severe depressive symptoms,severe physi-cal illness,history of being abused,afraid of being attacked,suicidal history of relatives or associates,severe of hopeless.Conclusion:The hotline counselors should be aware of the necessity of assessing the severity of depres-sion and other suicide-related factors for the callers with mental disorder diagnoses before providing any psychologi-cal intervention.Moreover,except for providing interventions on the callers'psychiatric problems,the hotline coun-selors should also identify and help to deal with the stress events in their lives.

OBJECTIVE: To explore the relationship between hypoxia-inducible foctor-1α (HIF-1α) and neovascularization in early atherosclerosis plaques by establishing rabbit carotid atherosclerosis models, and to observe the value of contrast-enhanced ultrasound in the detection of neovascularization. METHODS: We provided high-fat diet combined with the implantation of silicone rubber ring to establish carotid atherosclerosis in rat models. On the 14th and 28th days, we detected neovascularization in the carotid atherosclerotic plaques by contrast-enhanced ultrasound, obtained the peak intensity (PI) of the contrast agent in the plaques by time-intensity curve (TIC) and analyzed the difference. We also tested the level of HIF-1α, vascular endothelial growth factor (VEGF), cluster of differentiation 31 (CD31), α-actin, and RAM-11 by immunohistochemical method in each group, analyzed their correlation, and the correlation between PI and CD31 expression. RESULTS: On the 14th day, contrast-enhanced ultrasound showed the neovascularization in the carotid atherosclerotic plaques. On the 14th and 28th days, the intensity of contrast-enhanced ultrasound showed significant difference, the mean optical density of HIF-1α, VEGF, CD31, RAM-11, and α-actin within the carotid atherosclerotic plaques also showed statistical difference. The expressions between HIF-1α and VEGF, HIF-1α and CD31, HIF-1α and RAM-11, HIF-1α and α-actin, as well as PI and CD31 showed highly positive correlations. CONCLUSION During the process of atherosclerosis evolution, neovascularization in the atherosclerotic plaques has come into being in the early period, and HIF-1α in early atherosclerosis can promote the formation of neovascularization. Contrast-enhanced ultrasound can detect the dynamic changes of neovascularization within early atherosclerotic plaques.
Animals ; Carotid Artery Diseases ; complications ; metabolism ; pathology ; Disease Models, Animal ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Male ; Neovascularization, Pathologic ; etiology ; metabolism ; pathology ; Rabbits

Objective:To investigate the role of down-regulating serine racemase (SRR) in alleviating the β-amyloid peptide (Aβ) induced neurotoxicity and synaptic damage and possible mechanism in PC12 cells.Methods:(1) PC12 cells cultured in vitro were divided into 0, 20, 40 and 80 μmol/L Aβ 25-35 treatment groups; they were treated with 0, 20, 40 and 80 μmol/L Aβ 25-35 for 24 h, respectively; cell counting kit (CCK)-8 was used to detect the survival rate of cells in each group, and Western blotting was used to detect the SRR protein expression. PC12 cells were treated with 40 μmol/L Aβ 25-35 for 0, 12, 24 and 48 h, respectively; cell survival and SRR protein expression were detected by CCK-8 and Western blotting, respectively. (2) PC12 cells were divided into control group, nonsense sequence group, SRR small interfering RNA (siRNA) group 1, SRR siRNA group 2, and SRR siRNA group 3; cells in the later three groups were transfected with SRR nonsense sequence or different SRR siRNA sequences, respectively; 48 h after that, Western blotting was used to detect the SRR protein expression of cells in each group, and SRR siRNA with best effect was selected for subsequent experiments. (3) PC12 cells were divided into control group, AD group, AD+nonsense sequence group, and AD+SRR siRNA group; cells in the latter two groups were transfected with nonsense sequence or SRR siRNA for 48 h, respectively; cells in the latter three groups were added 40 μmol/L Aβ 25-35, and cells in the control group were added same amount of solvent; 24 h after treatment, the SRR protein expression was detected by Western blotting, cell survival was detected by CCK-8, cell apoptosis was detected by Hoechst 33258 fluorescent staining, Caspase 3 activity was detected by enzyme linked immunosorbent assay, and the expressions of activated Caspase 3, N-methyl- D aspartate (NMDA) receptor-associated proteins and postsynaptic dense protein 95 (PSD95) were detected by Western blotting. Results:(1) The survival rate of cells in 0, 20, 40 and 80 μmol/L Aβ 25-35 treatment groups was successively decreased and the SRR protein expression was successively increased, with significant differences ( P<0.05); PC12 cells treated with 40 μmol/L Aβ 25-35 for 0, 12, 24 and 48 h had successively decreased survival rate and successively increased SRR protein expression, with significant differences ( P<0.05). (2) The SRR protein expressions in the SRR siRNA group 1, SRR siRNA group 2 and SRR siRNA3 group 3 were significantly decreased as compared with those in the control group and nonsense sequence group ( P<0.05), and the decrease in the SRR siRNA group 2 was the most obvious. (3) As compared with the control group, the cells in the AD group had significantly increased SRR protein expression and apoptosis rate, statistically decreased cell survival rate, significantly increased Caspase 3 activity and activated Caspase 3 protein expression, significantly increased protein expressions of NMDA receptor 2A (NMDAR2A) and NMDA receptor 2B(NMDAR2B), and statistically decreased PSD95 protein expression ( P<0.05); as compared with cells in the AD group, cells in the AD+SRR siRNA group had significantly decreased SRR protein expression and apoptosis rate, statistically increased cell survival rate, significantly decreased Caspase 3 activity and activated Caspase 3 protein expression, significantly decreased NMDAR2A protein expression, and statistically increased PSD95 protein expression ( P<0.05). Conclusion:Down-regulation of SRR expression can reduce the NMDAR2A protein expression, alleviate the over-activation of NMDA receptor, reduce the cell apoptosis, improve cell survival rate, protect nerve cells, increase PSD95 protein expression, and alleviate synaptic damage in PC12 cells.

PURPOSE: An older female predominance has been reported among chronic cough patients in Western countries, which is considered to be associated with a higher cough sensitivity in females. However, the characteristics of Chinese chronic cough patients remain unclear. This study aimed to explore the age and sex distribution as well as their relationship with cough reflex sensitivity to capsaicin in Chinese chronic cough patients. METHODS: We analyzed the demographic features of 1,882 consecutive chronic cough patients who attended our cough clinic in Guangzhou, China. Cough sensitivity to capsaicin, which was defined as the lowest concentration of capsaicin causing 5 coughs or more (C5), was measured in 539 of the 1,882 patients and 68 healthy volunteers. RESULTS: The mean age of the patients was 43.0 ± 13.7 years and patients aged <50 years accounted for more than two-thirds of the study population. Around 87% of the patients were never-smokers. The proportion of females (51.5%) was almost equal to that of males (48.5%). The pattern of the age and sex distribution was consistently reflected within most common causes of chronic cough, while a female predominance was shown in patients with cough-variant asthma and patients aged ≥50 years. Female patients had higher cough sensitivity to capsaicin than male patients (log C5: 1.58 ± 0.84 vs. 2.04 ± 0.84 μmol/L, P = 0.001), and patients aged ≥50 years had higher cough sensitivity to capsaicin than patients aged <50 years. CONCLUSIONS: In China, patients with chronic cough have a roughly equal sex distribution and a middle-aged predominance, irrespective of a higher cough sensitivity to capsaicin in females and older patients. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT02591550
Age Distribution ; Asian Continental Ancestry Group ; Asthma ; Capsaicin ; China ; Cough ; Female ; Healthy Volunteers ; Humans ; Male ; Reflex ; Sex Distribution

Objective:To explore the current status of surgery for portal hypertension to grasp current status and future development of surgery in China.Methods:This study is jointly sponsored by China Hepatobiliary & Pancreatic Specialist Alliance & Portal Hypertension Alliance in China (CHESS).Comprehensive surveying is conducted for basic domestic situations of surgery for portal hypertension, including case load, surgical approaches, management of postoperative complications, primary effects, existing confusion and obstacles, liver transplantation(LT), laparoscopic procedures and transjugular intrahepatic portosystemic shunt(TIPS), etc.Results:A total of 8 512 cases of portal hypertension surgery are performed at 378 hospitals nationwide in 2021.Splenectomy plus devascularization predominated(53.0%)and laparoscopy accounted for 76.1%.Primary goal is preventing rebleeding(67.0%) and 72.8% of hospitals used preventive anticoagulants after conventional surgery.And 80.7% of teams believe that the formation of postoperative portal vein thrombosis is a surgical dilemma and 65.3% of hospitals practiced both laparoscopy and TIPS.The major reasons for patients with portal hypertension not receiving LT are due to a lack of qualifications for LT(69.3%)and economic factors(69.0%).Conclusions:Surgery is an integral part of management of portal hypertension in China.However, it is imperative to further standardize the grasp of surgical indications, the handling of surgical operation and the management of postoperative complications.Moreover, prospective, multi-center randomized controlled clinical studies should be performed.

OBJECTIVE To investigate the anti-pulmonary fibrosis effect of linarin in vivo and in vitro， and investigate its mechanism preliminarily. METHODS C57BL/6J mice were randomly divided into normal group （carboxymethylcellulose sodium）， model group （carboxymethylcellulose sodium）， positive control group （pirfenidone， 200 mg/kg）， linarin low-dose and high-dose groups （12.5， 25 mg/kg）， with 8 mice in each group. Except for normal group， pulmonary fibrosis model was induced in other groups. After modeling， they were given relevant medicine intragastrically， once a day， for consecutive 14 d. The general situation of mice was observed， and their lung indexes were measured； the levels of tumor necrosis factor-α （TNF-α） and transforming growth factor-β1（ TGF-β1） in serum and interleukin-6 （IL-6） in lung tissue were determined. Hematoxylin-eosin （HE） staining and Masson staining were used to observe the histopathological morphology of lung. The pulmonary fibrosis was scored according to Ashcroft score standard. The expressions of α-smooth muscle actin （α-SMA） and （type Ⅰ collagen， Collagen Ⅰ）， phosphorylated extracellular signal-regulated kinase （p-ERK1/2） and TGF-β1 in lung tissues were detected. HFL1 cells were stimulated by TGF- β1 to form pulmonary fibrosis model in vitro， which were divided into normal group， model group and linarin low-， medium- and high-concentration groups （3.7， 7.4， 14.8 mg/L）. After being cultured for 48 h， the protein expressions of α-SMA， Collagen Ⅰ and p-ERK1/2 in HFL1 cells were detected. RESULTS In vivo， compared with normal group， the lung index of model group and the levels of TNF- α， TGF- β1 and IL-6 were significantly increased （P＜0.01）. There were a large number of inflammatory infiltration and cellular fibrosis lesions in the alveoli， and a large number of collagen depositions. The scores of HE staining and Masson staining were significantly increased （P＜0.01）. The protein expressions of α-SMA， Collagen Ⅰ， p-ERK1/2 and TGF-β1 in lung tissue were up-regulated significantly （P＜0.01）. Compared with model group， above indexes of mice were improved significantly in linarin high-dose group （P＜0.05 or P＜0.01）， and most of indexes （except for lung index） were improved significantly in linarin low-dose group （P＜0.05 or P＜0.01）. In vitro， compared with blank group， the density of cells in the model group increased， and obvious proliferation and other changes occurred； protein expressions of α-SMA， Collagen Ⅰ and p-ERK1/2 were significantly up-regulated （P＜0.05 or P＜0.01）. Compared with model group， the cell density of each concentration group was decreased and the morphology gradually returned to normal； the expressions of above proteins in linarin high-concentration group and the protein expression of p-ERK1/2 in linarin medium-concentration group were down-regulated significantly（P＜0.05 or P＜0.01）. CONCLUSIONS Linarin may regulate ERK and inflammatory pathways to reduce the inflammatory response， thereby exerting anti-pulmonary fibrosis effect.