Objective:To establish a determination method for the content of cyproheptadine hydyochloride tablets and the related substances in the tablets by HPLC. Methods:The assay was performed on a CAPCELL PAK C18(Shiseido)(250 mm ×4.6 mm, 5μm) column with methanol-0. 002 5mol·L-1 sodium heptanesulfonate (adjusting pH to 3 with phosphoric acid)(60: 40) as the mo-bile phase. The detection wavelength was 225 nm, the flow rate was 1. 0 ml·min-1 , the column temperature was 30℃ and the sample size was 10 μl. Results: Cyproheptadine hydyochloride had good linear relationship within the range of 4. 12-82. 40 μg·ml-1 ( r=1. 000 0), and the average recovery was 99. 2%(RSD=0. 8%, n=9). The peaks of the related substances were well separated from that of cyproheptadine hydrochloride. Conclusion:The method is simple, fast and accurate, and can be used for the quality control of cyproheptadine hydyochloride tablets.

Objective To observe the control efficacy of community health education on population with metabolic syndrome (MS).Methods 56 cases of metabolic syndrome patients treated in the community were selected as the research object from June 2012 to 2013,and were randomly divided into the observation group (30 cases) and the control group (26 cases).The patients in the observation group received systematic community health education,and patients in the control group received regular treatment.The blood pressure,BMI Level,abdominal circumference,FPG level,TC,TG,LDL,HDL,2hPG,and HbAlc level of patients from both groups were compared one year later.Results Compared with the result before treatment,the abdominal circumference,blood pressure,blood glucose and lipid levels of patients in the observation group were all superior to the control group; the reasonableness for food taken was compared one year later,the observation group was also superior to the control group.Statistical significancc cxistcd in the differences between both groups.Conclusions The implementation of community health education has a positive significance in promoting the rehabilitation of people with metabolic syndrome,which is worth being promoted.

AIM:To investigate the effect of MK-2206, an inhibitor of protein kinase B (Akt), on the DNA damage of SGC-7901 cells.METHODS: SGC-7901 cells were treated with different concentrations of MK-2206, and phosphorylated histone H2AX (γ-H2AX) foci formation was detected by immunofluorescence staining .Western blot analy-sis was used to exam the levels of DNA damage-related protein.The expression of LC3-Ⅱ was determined to evaluate the change of autophagy .RESULTS:MK-2206 treatment increased the formation of γ-H2AX foci and histone H2AX phospho-rylation in the SGC-7901 cells.The levels of DNA damage response protein were also increased .In addition, MK-2206-treated SGC-7901 cells increased the expression of LC 3-II, a hallmark of autophagy .Inhibition of autophagy significantly enhanced MK-2206-mediated histone H2AX phosphorylation.CONCLUSION:MK-2206 induces DNA damage and auto-phagy in SGC-7901 cells.Blocking autophagy potentiates the response of MK-2206-induced DNA damage .

[ ABSTRACT] AIM:To investigate the biological characteristics of newborn rabbit tracheal chondrocytes in vitro. METHODS:Newborn rabbit tracheal chondrocytes were obtained by the method of enzyme digestion, and then cultured in monolayer in vitro.Morphological and growth observations were performed under inverted phase contrast microscope.The ultrastructures of the cells were observed under scanning electron microscope and transmission electron microscope.The bi-ological characteristics of secreted extracellular matrix components were detected by real-time PCR, immunocytochemistry staining and toluidine blue staining.RESULTS: Newborn rabbit tracheal chondrocytes isolated and cultured in vitro showed short triangular or irregular shapes, and adherent growth very well.The ultrastructures of the cells showed pore and abundant cytoplasm and organelles, with a lot of protein secretions in the cells.The chondrocytes expressed the mRNA of collagen I, collagen II and proteoglycans, mainly collagen II and proteoglycans.Immunocytochemistry staining showed col-lagen II and SOX9 positive, and collagen I weakly positive.Toluidine blue staining was also positive.CONCLUSION:Enzyme digestion and monolayer culture are suitable method to obtain newborn rabbit tracheal chondrocytes.These cells, secreting extracellular matrix components, are able to be selected as seed cells for tissue engineering of trachea in vitro, and used to study the therapeutic method for neonatal rabbit tracheal stenosis.

BACKGROUND: Bone marrow mesenchymal stem cells are important seeded cells for construction of tissue-engineered trachea, but there is no special surface marker. Therefore, identification of bone marrow mesenchymal stem cells is mostly based on morphology, phenotype antigen and the function of differentiation. OBJECTIVE: To explore the feasibility of the tracheal chondrogenic differentiation of bone marrow mesenchymal stem cells under a special condition through isolation, cultivation and identification of bone marrow mesenchymal stem cells. METHODS: Rabbit bone marrow was acquired in the sterile environment to isolate and culture bone marrow mesenchymal stem cells to passage 2 by bone marrow adherence and screening method. Flow cytometry identified the phenotype CD44, CD45 of bone marrow mesenchymal stem cells at passages 1 and 2. Rabbit tracheal samples were acquired in the sterile environment, the tracheal chondrocytes were isolated and cultured by enzyme digestion, and toluidine blue staining was used to detect aggrecan. Bone marrow mesenchymal stem cells were co-cultured with tracheal chondrocytes by Transwel and transforming growth factor β1. Cel morphology was detected under an inverted microscope. Real-time quantitative PCR and toluidine blue staining detected the extracel ular matrix components, such as type Ⅱ col agen and aggrecan.RESULTS AND CONCLUSION: After isolation and culture, cells were spindle and irregular in morphology, and passaged cells thrived that were gathered into a fish-like colony growth. For passage 1 bone marrow mesenchymal stem cells, the positive rates of phenotype antigen CD44 and CD45 were respectively 96.97% and 13.72%; for passage 2 cells, the positive rates of phenotype antigen CD44 and CD45 were 99.11% and 8.54%, respectively. Tracheal chondrocytes were positive for toluidine blue staining. The morphology of induced bone marrow mesenchymal stem cells changed from long fusiform to triangular or irregular shape, indicating the chondrocytes expressed type Ⅱ col agen and aggrecan, and toluidine blue staining was positive. These results showed bone marrow adherence and screening method could acquire bone marrow mesenchymal stem cells, and the purity of passage 2 cells is higher. Under a special condition, bone marrow mesenchymal stem cells have the potential of chondrogenic differentiation, and can be selected as seed cells for construction of tissue-engineered trachea.

Objective To investigate the correlation between the indications,findings,interventions of fibrobronchoscopy(FB) in neonates and their correlative diseases with neonatal FB results and clinical data.Methods Retrospective case series of 243 consecutive patients of 28 days old or younger were investigated underwent FB for the first time from January 2010 to December 2014,at a tertiary care hospital.The common indications for FB and detection rate of respiratory tract diseases were collected.If the findings of FB had significant associations with premature birth and other diseases were analyzed.Associations between interventions and basic illnesses were also analyzed.Results Of the 243 patients undergoing 275 procedures of FB,201 cases were boys(73.1％).The age of FB was (13.34 ± 9.76) days and the weight was (3.08 ± 0.68) kg.Forty-five cases were premature infants (16.4％).A total of 254 procedures were found to have congenital diseases (92.4％),and 177 cases of them had congenital heart diseases (CHD) (64.4％).Common indications for FB were dyspnea(140 cases,50.9％),tachypnea(82 cases,29.8％),and stridor(71 cases,25.8％).A total of 188 upper airway lesions were found and the most common findings were laryngomalacia(56 cases,20.4％) and vocal cord paralysis(bilateral/unilateral,50 cases,18.2％).A total of 315 lower airway lesions were found and the most common findings were airway mucosal inflammation (98 cases,35.6％),trachea and main bronchial stenosis (73 cases,26.5 ％).A total of 21 cases (7.6％) underwent supraglottoplasty during or after FB,while 17 cases (6.2％) underwent tracheal dilation and 10 cases (3.6％) underwent tracheotomy.Compared with non-CHD neonates,neonates with CHD were statistically significantly less likely to have congenital lesions statistically,such as laryngomalacia(15.8％ vs.28.6％,P =0.012),bilateral vocal cord lesions(6.2％ vs.21.4％,P =0.000) and congenital laryngeal dysplasia(0 vs.7.1％,P =0.001).The tracheotomy(0 vs.10.2％,P =0.000) and supraglottoplasty(2.3％ vs.17.3％,P =0.000) were more rare.Nevertheless,they were more likely to have secondary lesions such as the left main bronchial stenosis caused by extrinsic compression (23.7％ vs.1.0％,P =0.000),abnormal bronchial anatomy(9.6％ vs.2.0％,P =0.018),left vocal cord paralysis(9.0％ vs.1.0％,P =0.008) and airway mucosal inflammation(41.8％ vs.24.5％,P =0.004).The tracheostenosis and main broncial stenosis (37.3％ vs.7.1％,P =0.000) with long-term intubation(78.5％ vs.58.2％,P =0.000) were more common.There was no significant difference between term neonates and premature infants in the detection rate of respiratory tract diseases (P ＞ 0.05),tracheotomy (0 vs.4.3 ％,P =0.322),supraglottoplasty (13.3 ％ vs.6.5 ％,P =0.205) or long-term i ntubation (80.0％ vs.69.6％,P =0.157).Complications caused by procedure were rare and mild.Conclusions FB can detect whether the neonates with dyspnea,tachypnea and stridor have laryngomalacia,vocal cord paralysis,airway mucous edema,tracheal and main bronchial stenosis and other signs,and FB may play an important role in diagnosis,treatment and prognosis evaluation of neonatal respiratory diseases.

Objective: High mobility group box 3 (HMGB3) plays an important role in the development of various cancer. This study aims to explore whether HMGB3 regulates cervical cancer (CC) progression and elucidate the underlying mechanism. Methods: HMGB3 expression in clinical patients' tumor samples were determined by real-time quantitative polymerase chain reaction (qRT-PCR) and western blot. HMGB3 overexpression/knockdown were used to investigate its function. Cell apoptosis and cycle were detected by Annexin V/PI staining and flow cytometry. In vivo tumor model was made by subcutaneous injection of HeLa cells transfected with shRNAs targeting HMGB3 (shHMGB31) into the flank area of nude mice. Western blot was used to detect the levels of β-catenin, c-Myc, and matrix metalloproteinase-7 (MMP-7) in Hela and CaSki cells transfected with sh-HMGB3 or shRNAs targeting β-catenin. Results: Both messenger RNA and protein levels of HMGB3 were upregulated in CC tissues from patients. High expression level of HMGB3 had positive correlation with serosal invasion, lymph metastasis, and tumor sizes in CC patient. Functional experiments showed that HMGB3 could promote CC cell proliferation both in vitro and in vivo. The expression levels of c-Myc and MMP-7 were increased, resulting in regulating cell apoptosis, cell cycle, and activating Wnt/β-catenin pathway. Conclusions Our data indicated that HMGB3 may serve as an oncoprotein. It could be used as a potential prognostic marker and represent a promising therapeutic strategy for CC treatment.

OBJECTIVETo explore the effect of recombinant human soluble CD(40) ligand (rhsCD(40)L) and CD(40)L cDNA transfected cell (CD(40)L-TC) on the behavior of malignant B lymphocytes, and investigate the possibility of using rhsCD(40)L as a new bio-factor in tumor immunotherapy.

METHODrhsCD(40)L and CD(40)L-TC were obtained by gene recombinant techniques. Multiple myeloma cell lines, XG2, XG7, U266 and 8226, B-lymphoma cell lines, Raji and Daudi were selected to detect responses to rhsCD(40)L and CD(40)L-TC stimulation. Cell growth curve, cell cycle, early apoptosis as well as membrane surface molecules on these cell lines were analyzed.

RESULTS(1) The expression levels of CD(40) molecule on malignant B lymphocytes showed heterogeneity. High level of CD(40) on XG2, moderate on 8266, Raji, and Daudi, and no expression on U266 and XG7 were detected. The rhsCD(40)L stimulation gave rise to a typical homo-type cell aggregation of XG2 and Daudi. Meanwhile, at least 10 to 20 of CD(40)(+) XG2 or CD(40)(+) Daudi cells were found adherent to one pre-treat ed CD(40)L-TC. (2) Co-incubation with rhsCD(40)L (5 micro g/ml), or CD(40)L-TC (tumor cell: CD(40) = 5:1) resulted in a significant inhibition of in vitro cell growth of XG2, Raji and Daudi, with G(1)-phase arrest for XG2 and G(2)-phase for Raji and Daudi. These two kinds of CD(40) stimulators induced XG2, Raji and Daudi cells to apoptosis in vitro. The apoptotic rate for XG2 was 23.3% (rhsCD(40)L) and 18.8% (CD(40)L-TC), for Daudi 14.2% and 15.9%, and for Raji 11.6% and 8.9% respectively. (3) Phenotype analysis showed that CD(95) expression levels were significantly up-regulated on XG2, Raji and Daudi after stimulation with rhsCD(40)L or CD(40)L-TC, and CD(80) and CD(18) expression levels on Raji were respectively enhanced and decreased.

CONCLUSIONThe abilities to directly inhibit XG2, Daudi and Raji cell proliferation, to induce themapoptosis, as well as to up-regulate immune co-stimulator molecule CD(80) expression on Raji cells would make rhsCD(40)L a potential bio-factor for tumor immuno-therapy.

B-Lymphocytes ; drug effects ; metabolism ; pathology ; CD40 Antigens ; metabolism ; CD40 Ligand ; genetics ; metabolism ; pharmacology ; Cell Division ; drug effects ; Coculture Techniques ; DNA, Complementary ; genetics ; Humans ; Lymphoma, B-Cell ; metabolism ; pathology ; Recombinant Proteins ; pharmacology ; Time Factors ; Transfection ; Tumor Cells, Cultured ; drug effects

Objective:To analyze the value of minute ventilation to carbon dioxide production slope (VE/VCO 2 slope) combined with peak systolic blood pressure (SBP) in predicting prognosis for patients with chronic heart failure (CHF). Methods:A total of 170 patients with CHF who visited the Cardiac Rehabilitation Center of Tongji Hospital Affiliated to Tongji University and completed cardiopulmonary exercise test from March 2007 to December 2018 were enrolled in the study. The clinical data, cardiopulmonary exercise testing results and follow-up information of patients were collected to explore the predictors of all-cause mortality in patients with CHF.Results:The median follow-up time was 647 (182-1 764) days. All-cause death occurred in 34 patients. Compared with surviving patients, the proportion of diabetes and angiotensin-converting enzyme inhibitor/angiotensin Ⅱ receptor blocker (ACEI/ARB) use in fatal patients was significantly higher ( P<0.01). The VE/VCO 2 slope and peak SBP*VE/VCO 2 in the fatal patients were significantly higher, and the peak oxygen consumption (peak VO 2) was lower than those in the surviving patients ( P<0.01). The areas under the receiver operating characteristic curve (AUC) of VE/VCO 2 slope and peak SBP*VE/VCO 2 in predicting all-cause mortality in patients with CHF were 0.648 ( P=0.008) and 0.681 ( P=0.001), respectively; the optimal thresholds were >40.95 ( P=0.008) and > 5 423.50 mmHg (1 mmHg=0.133 kPa, P=0.006), the sensitivity was 0.559 and 0.588, and the specificity was 0.728 and 0.735, respectively. Multivariate Cox regression analysis showed that after adjusting for age, gender, diabetes and ACEI/ARB use, VE/VCO 2 slope ( HR=2.12, P=0.036) and peak SBP*VE/VCO 2 ( HR=2.42, P=0.016) were independent risk factors for all-cause mortality in patients with CHF. Conclusion:Compared to the traditional index VE/VCO 2 slope, a novel index peak SBP* VE/VCO 2 provides a relatively better predictive value for all-cause death of CHF patients.

ObjectiveTo study the traditional Chinese medicine （TCM） syndromes of children with alopecia areata， and provide evidence for TCM differentiation and treatment in clinic. MethodsA retrospective analysis was conducted on the clinical data of 800 children with alopecia areata admitted to the Hair Medicine Center of the China-Japan Friendship Hospital from January 1， 2012 to December 31， 2021. The clinical data of the children were collected using a four-examination information questionnaire， including clinical characteristics （age of consultation， age of onset， course of disease， family history， severity grading）， alopecia areata-related factors （triggers）， and four-examination information （including sleep， diet， emotions， bladder and bowel function， etc.）. Descriptive frequency analyses， rank sum tests， factor analyses and cluster analyses were performed， and the distribution of the major TCM syndromes was summarised with the clinical data. ResultsThere were 800 children with alopecia areata， including 449 males and 351 females； 8 cases （1.00%） were in infancy， 36 cases （4.50%） were in early childhood， 180 cases （22.50%） were in preschool， 380 cases （47.50%） were in school age， and 196 cases （24.50%） were in puberty at the time of consultation； the average age of consultation was 8.31±3.86 years， the average age of onset of disease was 5.40±3.82 years， and the average duration of disease was 2.94±2.77 years； 527 children （65.87%） with severe alopecia areata； 85 children （13.56%） had a family history of alopecia areata； 772 children （96.50%） had unknown triggers for their first alopecia areata， and 28 children （3.50%） reported the presence of obvious triggers， including fright （9 cases）， high fever （5 cases）， allergic reactions （4 cases）， micronutrient （zinc， iron， etc.） deficiencies （4 cases）， inappropriate diet （2 cases）， environmental factors （1 case， new house renovation）， atopic dermatitis （1 case）， atopic asthma （1 case）， and pneumonia （1 case）. A total of 40 four-examination information items were collected， among which the frequency of kicking quilts was the highest with 380 cases （47.50%）， followed by picky eating （369 cases， 46.13%）， sleeplessness （334 cases， 41.75%）， irritability （334 cases， 41.75%）， partiality towards certain foods （306 cases， 38.25%）， impulsiveness （297 cases， 37.13%）， dry stools （233 cases， 29.13%）， yellow urine （215 cases， 26.88%）， nail biting （213 cases， 26.63%）， bad breath （211 cases， 26.38%）. According to factor analysis and cluster analysis， five types of TCM syndromes were obtained， in order as qi and blood deficiency syndrome （110 cases， 13.75%）， spleen deficiency syndrome （114 cases， 14.25%）， kidney essence deficiency syndrome （140 cases， 17.50%）， dietary stagnation syndrome （150 cases， 18.75%）， and liver depression and spleen deficiency syndrome （286 cases， 35.75%）. Patients in each age group and SALT grading are mainly liver depression and spleen deficiency syndrome. ConclusionThe TCM symptoms of children with alopecia areata are mainly based on qi and blood deficiency syndrome， spleen deficiency syndrome， kidney essence deficiency syndrome， dietary stagnation syndrome， and liver depression and spleen deficiency syndrome， of which liver depression and spleen deficiency syndrome is the most common type at different ages and stages of the disease.