OBJECTIVE: This study aims to establish an effective and stable periodontal ligament cell line stably expressing human telomerase reverse transcriptase (hTERT) gene by using the adenovirus method. METHODS: Polymerase chain reaction (PCR) was used to amplify the full length of hTERT gene to construct recombinant adenovirus plasmid pAd-pshuttle-cmv-hTERT. Packaged adenovirus particles were used for infection of human periodontal ligament cells. The expression levels of hTERT and osteogenic genes, such as alkaline phosphatase, Runt-related transcription factor 2, bone sialoprotein, osteocalcin, osteopontin, and collagen Ⅰ mRNA, were detected by quantitative real-time PCR (qRT-PCR). The ability of osteogenic differentiation was observed by alizarin red staining, and the cell proliferation was determined by CCK-8. RESULTS: Adenovirus particles containing the hTERT gene were successfully constructed and infected with periodontal ligament cells. The infected cells were similar to normal periodontal ligament cells. The qRT-PCR results showed that hTERT and osteogenesis-associated genes were highly expressed in the periodontal ligament cell lines constructed by adenoviruses. Alizarin red staining showed that the periodontal ligament cell line had strong osteogenic differentiation capability. CCK-8 showed that the periodontal ligament cell line had strong proliferation capability. CONCLUSIONS The human periodontal ligament cell line with high efficiency and stable expression of hTERT was established by the adenovirus method, thereby providing an ideal cell line for studying the mechanism of periodontal regeneration.
Adenoviridae ; Alkaline Phosphatase ; Cell Differentiation ; Cell Line ; Cell Proliferation ; Humans ; Osteogenesis ; Periodontal Ligament ; Telomerase

Adolescent ; Adult ; Aged ; Antidepressive Agents ; therapeutic use ; Body Mass Index ; Depression ; microbiology ; Diarrhea ; microbiology ; Duloxetine Hydrochloride ; therapeutic use ; Feces ; microbiology ; Humans ; Irritable Bowel Syndrome ; drug therapy ; microbiology ; Middle Aged ; Pilot Projects ; Probiotics ; therapeutic use ; RNA, Ribosomal, 16S ; genetics ; Young Adult

OBJECTIVE: To investigate the changes of mean platelet volume (MPV), platelet distribution width (PDW) and platelet associated antibodies (PAIg) in children with acute immune thrombocytopenic purpura (aITP), and to explore the diagnostic value of MPV, PDW, PAIg and their combination for megakaryocyte dysmaturity in aITP children. METHODS: Plt count, MPV and PDW of 36 aITP children were measured by using Sysmex XN automatic blood cell analyzer, and 33 children with acquired thrombocytopenic purpura (ATP) without megakaryocyte dysmaturity. The expression of PAIg was detected by flow cytometry, and the number and classification of megakaryocytes in the bone marrow were performed by marrow cytology. The diagnostic significances of MPV, PDW, PAIg and their combination as well as the sensitivity and specificity for megakaryocytes dysmaturity in aITP were assessed through calculating the area under ROC curve (AUC), after determining the influence of each parameters on the megakaryocyte dysmaturity by Logistic regression. RESULTS: MPV, PDW and PAIg of aITP children were significantly higher than those of the ATP children (P＜0.05), while the Plt count and number of thromocytogenic megakaryocytes per area (1.5 cm×3 cm) were less than those of the controls (P＜0.05). Count of RBC and WBC, percentages of neutrophil granulocytes and lymphocydes in aITP were similar to those in the controls（P＞0.05）. The results of Logistic regression showed that Plt count, MPV, PDW and PAIg were the factors influencing megakaryocyte dysmaturity in aITP children, and the regression model has a high statistical significance (χ=65.491，P=0.001) and r square (R=0.713). The AUC of the combined detection of Plt count, MPV, PDW and PAIg was 0.863, which was much higher than that of Plt count, MPV, PDW, PAIg individually or in pairs. The sensitivity and specificity of the combined detection were 79.167% and 89.697%, which were higher than those of Plt count, MPV, PDW, PAIg individually or in pairs. CONCLUSION The diagnostic significance of MPV and PDW for megakaryocyte dysmaturity in aITP are insufficient, but the diagnostic efficacy can be improved by combined examination with PAIg.
Antibodies ; Blood Platelets ; Child ; Humans ; Mean Platelet Volume ; Megakaryocytes ; Platelet Count ; Purpura, Thrombocytopenic, Idiopathic ; diagnosis

OBJECTIVE: To explore the effect and mechanism of miR-30b on cisplatin-resistance of human NK/T cell lymphoma lines SNK-6 and YTS cells. METHODS: Normal NK cells, SNK-6 and YTS cells were cultured, the expression levels of miR-30b and macrophage-derived chemokine (CCL22) were detected by real-time PCR assay, and the CCL22 expression was detected by Western blot. The SNK-6 and YTS cells were transfected with miR-30b mimics and inhibitor respectively, then the effect of cisplatin resistance in SNK-6 and YTS cells was measured by MTT assay, the activity of caspase-3 was detected by caspase-3 assay kit, and the cell apoptosis was analyzed by flow cytometry. Dual-luciferase reporter gene assay was used to determine the targeting relationship between miR-30b and CCL22. Furthermore, the effect of CCL22 on cisplatin-resistance and caspase-3 actirity was also evaluated. RESULTS: Compared with the normal NK cells, the expression levels of miR-30b significantly decreased in both SNK-6 and YTS cells (P＜0.01), but CCL22 mRNA expression increase in both cells (P＜0.01). MiR-30b mimics decreased the cell activity (P＜0.05), down-regulated the cisplatin-resistance (P＜0.05), and increased cell apoptosis and caspase-3 activity (P＜0.05). The effects of miR-30b inhibitor were contrary to the mimics. Up-regulation of miR-30b expression significantly decreased the luciferase activity in CCL22 3'-UTR-transfected NK cells, but not in Mut-CCL22 3'UTR group, suggesting that CCL22 could act as a direct target of miR-30b. The expressions of CCL22 pathway proteins were down-regulated after SNK-6 cells transfected with miR-30b mimics (P＜0.05), while this effect was restored by overexpression of CCL22. Moreover, CCL22 overexpression also increased the cell activity and decreased caspase-3 activity when SNK-6 cells were transfected with miR-30b mimics. CONCLUSION MiR-30b inhibits cisplatin-resistance of human NK/TCL SNK-6 and YTS cells by targeting CCL22.
Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Chemokine CCL22 ; Cisplatin ; Gene Expression Regulation, Neoplastic ; Humans ; Killer Cells, Natural ; Lymphoma, T-Cell ; genetics ; MicroRNAs ; T-Lymphocytes

Objective To indentify the cognitive status of Chinese patients to acne and the influencing factors to theirs' cognitive status,so as to provide solid evidences for the prevention and treatment of acne.Methods A self-designed questionnaire was made to conduct this survey of 16,156 acne patients,who seeked to the treatment in the dermatological departments from 112 hospitals in China.The survey consisted of several parts,including the general status of patients,the patients' cognition of occurrence,development and risk factors of acne,whether the first choice was seeking treatment at the hospital when the patients had acne and the condition of selection of skin care products.The factors were analyzed,which could impact the cognition of the patients' behavior of treatment,how did the patients' cognition to influence their medical behavior and skin care as well as the consistency of assessment of the severity of acne by doctors and patients themselves.Results The acne patients studied had the best knowledge of "acne is a skin disease","it not only occurs in the period of adolescence" and "the disease can be prevented and cured",which accordingly accounted for 80.65％,69.16％ and 65.49％ of the total patients respectively.However,the awareness of acne patients to heredity,high sugar and dairy products as risk factors for acne was insufficient,which accounted for 48.72％,42.40％ and 18.25％ of the total patients,respectively.Gender,age,educational level,occupation and health knowledge were the main factors affecting the cognitive level of patients;the survey also found that men,patient with educational level of junior high or even lower educational condition,occupation of labor workers or farmers and patients were lack of health education with poor knowledge of the genetics and dietary were risk factors for acne;patients with age over 36 years or with mild illness had poor knowledge of dietary risk factors for acne;the difference was statistically significant (P＜0.05).The analysis of the influence of cognitive status on medical treatment behavior and skin care showed that the better the cognition,the higher the probability of patients would choose medical treatment as the first choice as well as choosing functional skin care products;the difference was statistically significant (P＜0.05).The consistency of assessment of the severity of acne by doctors and patients was poor (Kappa value ＜0.4),and the assessment of severity of acne by patients was more serious than doctors' assessment.Conclusions Patient's cognitive status will affect their medical behavior and skin care,and there is also a phenomenon that patients have a more serious assessment of their acne condition.It is suggested that health education for acne patients should be strengthened in clinical medicine so as to improve their knowledge of acne as well as preventing from acne effectively.

Diabetic nephropathy(DN) is a kind of severe diabetes mellitus(DM) complications,which is one of the most common cause of end-stage renal failure.The incidence of DN increases with the increase of the number of DM patients,and early prevention is the key.DM and DN are characteristic advantages of Tibetan medicine,the "holistic view" of dynamic balance based on the three major factors in the body(Sanyin,included rlung,mkhris-pa and bad-gan) is a unique understanding of DM and DN among Tibetan medical practitioners,and which has rich and unique clinical and medication experiences in DN treatment.This paper discussed the medical theory "holistic view" on the Tibetan medicine for DM and DN,dug commonly used Tibetan drugs and Tibetan medical prescriptions for clinical treatment of DN,in order to provide new ideas for the treatment of DN and the discovery of new drugs for DN, and give full play to the unique advantages of Tibetan medicine.

Objective: To optimize purification process of total alkaloid extract of Berberis dictyophylla cortex by macroporous resin,and to establish its quality standard. Method: Acid dye colorimetry was used to investigate the purification process of total alkaloid extract of B. dictyophylla cortex,the process parameters included concentration of sample solution,speed of sampling,diameter-height ratio of resin column,water washing amount,concentration and dosage of eluent,flow rate of elution,etc.In order to determine the optimum process,HPLC was employed to determine the contents of four alkaloids(magnoflorine,jatrorrhizine hydrochloride,palmatine hydrochloride,and berberine hydrochloride) with mobile phase of acetonitrile-0.1% phosphoric acid aqueous solution for gradient elution and detection wavelength at 270 nm.After being purified,quality standard of total alkaloid extract of B. dictyophylla cortex was investigated according to the requirements in the 2015 edition of Chinese Pharmacopoeia. Result: Optimal purification conditions were as following:10 g of HPD100 macroporous adsorption resin with a column diameter-height ratio of 1:8,sampling solution concentration of 11 g·L^-1,the loading flow rate of 1 mL·min^-1,sampling solution volume of 50 mL,washed with 4 BV of water(1 BV=15 mL) and added 9 BV of 30% ethanol,after being purified,the transfer rate of total alkaloids was>80%,and its purity was>65%.The quality standard of total alkaloid extract of B. dictyophylla cortex was established,there were 19 common peaks in the characteristic chromatogram,and the overall similarity was>0.99. Conclusion: This optimized purification process is stable and feasible, and the established quality standard is controllable.

Objective To observe the effect of breathing training on postpartum stress urinary incontinence (SUI) based on pelvic floor muscle exercise and biofeedback electrical stimulation. Methods From July to October, 2017, 64 SUI patients at the sixth week after childbirth were randomly divided into con-trol group (n=32) and observation group (n=32). The control group was treated with pelvic floor muscle exercise and biofeedback electrical stimulation. The observation group was supplemented with breathing training addition-ally. The pelvic floor muscle myoelectric value, urinary incontinence frequency, one hour urine pad test and the average thickness of the transversus abdominis were evaluated before and twelve weeks after treatment. Results Before treatment, there was no significant difference in the pelvic floor muscle myoelectric value, urinary incon-tinence frequency, one hour urine pad test and the average thickness of the transversus abdominis between two groups (P>0.05). Twelve weeks after treatment, the pelvic floor muscle myoelectric value significantly increased in both groups (t>12.564, P<0.001), and was higher in the observation group than in the control group (t=14.064, P<0.001); the urinary incontinence frequency and the positive rate of one hour urine pad test significantly de-creased in both groups (Z>9.615, P<0.001), and were lower in the observation group than in the control group (Z=-2.767, χ2=6.564, P<0.05); the average thickness of the transversus abdominis had no improvement in the control group (t=-1.510, P>0.05), and significantly increased in the observation group (t=-10.681, P<0.01), which was higher in the observation group than in the control group (t=7.541, P<0.001). Conclusion On the basis of pelvic floor muscle exercise and biofeedback electrical stimulation, supplementation with breathing training could reduce the symptoms of postpartum stress urinary incontinence, and improve the func-tion and the quality of life.

OBJECTIVETo study the clinical characteristics of patients with post-transplantation lymphoproliferative disease (PTLD) after allogeneic peripheral blood hematopoietic stem cell transplantation, and to improve the understanding and diagnosis of PTLD.

METHODSThe clinical data of 244 patients underwent allogeneic hematopoietic stem cell transplantation in the General Hospital of PLA from May 2014 to April 2017 were analyzed retrospectively. The follow-up time was up to November 30, 2017. The incidence, risk factors, treatment and survival of patients with PTLD were statistically analyzed.

RESULTSAmong the 244 cases the PTLD occurred in 22 cases, the incidence rate was 9.02%, 5 of them were diagnosed by pathology, and 17 were diagnosed clinically. All of them had EB virus infection. They were all ATG user, either underwent related haploidentical hematopoietic stem cell transplantation or unrelated hematopoietic stem cell transplantation, 20 cases were treated with rituximab or rituximab combined with γ-globulin, glucocorticoid, ERV+CTL, chemotherapy and 17 showed the effective response, with a total effective rate of 85%. The median follow-up time was 122 days, the median survival time was 5 months (1-22 months) and the total survival rate was 50%.

CONCLUSIONThe incidence of PTLD after allogeneic peripheral blood hematopoietic stem cell transplantation closely relates with EB virus infection. The application of ATG in the preconditioning scheme is a high risk factor for the onset of PTLD. In the case of no pathological diagnosis, clinical and laboratory examinations should be actively combined so as to define clinical diagnosis. The riturimab should be used more and more for patients with PTLD.

Objective To provide a besis of the control measures for foodborne disease outbreaks by analysing the laws and the popular features of foodlorne disease outbreaks in yunnan from 2013 to 2017.Methods The data of epidemic situation and field investigation of foodborne disease outbreaks from 2013 to 2017 in Yunnan province was analyzed. Results From 2013 to 2017,a total of 2 519 cases of foodborne disease outbreaks were repeatedin Yunnan Province,Among 18681 cases, 257 patients died with mortality 1.38% . Foodborne disease outbreaks occur in the third quarters.Outbreaking and outbreaking from a few number of households, followed by the work unit Employer Unit; outbreaking deaths in the family is also the most. Primary causal factor is plant. Pathogen the pathogenic bacteria main pathogen are Salmonella (38.89%), Bacillus cereus intoxication bacillus (38.89%),staphylococcus aureus (11.11%), Diarrheagenic escherichia coli (5.56%), Proteus Proteus species Proteus vulgaris (5.56%),et al. Conclusion From 2013 to 2017,foodborne disease outbreaking in yunnan is mainly due to plants and micro－organisms and mosty oceurs at home and canteens. It is necessary to strengthen the food hygiene management at home and canteens,to avoid foodborne disease outbreaking incidents.