Objective To compare the influence of three kinds of complete media with 0. 10 ,0. 15 ,0. 20 fetal bo-vine serum( FBS) on purity and cycle of rat bone marrow mesenchymal stem cells ( BMSCs) cultured in vitro and to seek suitable FBS concentration for the cultivation of the stem cells. Methods SD rats were executed by cervical dislocation method and used whole bone marrow adherence to isolate rat BMSCs. Experiment was divided into A, B,C,3 groups. Compared the expression of CD45, CD29, CD90, CD44 in the three groups in 2,3,4,5 passages ( P2 , P3 , P4 , P5 );the cells of P3 in group A were digested and cultured in three different concentration of com-plete culture media for four days, measuring cell cycle in 24,48,72,96 h by flow cytometry instrument. BMSCs of P3 were collected and inoculated to 6 pieces of 96-well plates, then vaccinated with complete media with 0. 10, 0. 15,0. 20 FBS in every plate, one culture plate was taken out for optical density(OD) meaturement every day with CCK-8. Results CD45 was negative, CD29, CD90, CD44 were positive. The difference of BMSCs surface markers cultured in the three kinds of complete media was bigger in the first two passages, but the difference was less in P3 , P4 , all could obtain pure BMSCs in P4 relatively;according to the results of the cell cycle at the same time, G0/G1 phase:with the increase of concentration of fetal bovine, G0/G1 phase reduced and had no diference in A, B, C groups;G2/M phase:there was difference between them after 24 h(P<0. 05,F=12. 412), but with the extension of time, the differences disappeared;S phase:there was no difference in the three groups;S+G2/M phase increased with the concentration of FBS. According to the result of cell vitality, the OD of ABC three groups increased in turn in 24 h, and there was difference(P<0. 05,F=5. 002), but with the extension of time, there was no obvious difference between them. Conclusion The three kinds of culture media in P4 can obtain pure BM-SCs, cell cycle and vitality show three complete media can promote the growth of BMSCs. There is no difference between them. Culture media with 0. 10 FBS can satisfy the isolation and amplification of BMSCs, in order to ob-tain pure BMSCs in the short term, using culture media with 0. 15 and 0. 10 FBS in the primary culture and subcul-ture respectively.

Objective To evaluate the standardized drug treatment and its influence factors of patients,with coronary artery disease,in hospital and one year after discharge.Methods The study enrolled sequentially 165 patients who were firstly diagnosed of coronary artery disease,61 cases with stable angina,67 cases with unstable angi-na and 37 cases of acute myocardial infarction,by coronary artery angiography from 2010 to 2012.The standardized drug treatment and its influence factors of patients were analyzed at hospital and 1 year after discharge in the present study.Results Fifty five percent patients with coronary artery disease at hospital regularly took the four drugs,anti-platelet agents,statins,beta blockers and angiotensin converting enzyme inhibitors/angiotensin II receptor antagonist. The ratio decreased to forty five percent one year after discharge(χ2 =81.04,P <0.01).The reasons of the irregular medication taken were optional withdrawal(61%),following the doctors′advice(15%),economic hardship(20%) and the drugs′adverse reaction(4%).Conclusion The results of the present study showed that the rate of regular medication in patients with coronary artery disease is low in hospital and 1 year after discharge.The major reasons of the irregular medicine taken were the patients′optional withdrawal and the consciousness of second prevention was lack in doctors in our hospital.Therefore,the increase of the consciousness of regular standardized treatment in prima-ry care physicians and strengthen the management of the patients after discharge form hospital.

Objective To compare the efficacy and safety of paroxetine alone and combined with folic acid in patients complaining of premature ejaculation ,and measured the 5-hydroxyptamine(5-HT ) concentration in two groups before and after treatment and compared the differences .Methods 126 cases of PE were included from department of Urology of 363 Hospital of Chengdu .Subjects were randomly divided into 2 groups ,group A were given paroxetine hydrochloride 20 mg/d ,group B were given paroxetine hydrochloride 20 mg/d and fo-lic acid 0 .4 mg/d ,study duration was 8 weeks .Blood sample got from the candidates both in screening period and after 8 weeks of treatment .The efficiency after treatment was measured by IELT and PEP ,the plasma 5-HT level was measured too .SPSS16 .0 statistical analysis was used .Results After treatment ,IELT of group A and group B was improved from 1 .21 ,1 .18 min to 8 .04 ,9 .42 min .The improvement of average IELT in group B was significantly higher than that in group A ,the difference was statistically significant (P＜0 .05) ;The PEP score and 5-HT level in group B were significantly higher than that in group A ,the difference was statistically significant (P＜0 .05) .Conclusion Paroxetine combined with folic acid in the treatment of primary premature ejaculation has a significant effect ,compared to paroxetine alone .The average plasma level of 5-HT increased significantly ,and folic acid could assist paroxetine in elevating plasma levels of 5-HT and improving primary premature ejaculation symptoms .

Objective:To comprehensively evaluate the correlation between migraine and the risk of hemorrhagic stroke using Meta-analysis.Methods:The published observational studies on migraine and the risk of hemorrhagic stroke in PubMed, EMbase, Cochrane library, Chinese Biomedical Database, China Journal Full-text Database, Wanfang Database and VIP Database were retrieved by computers. The retrieval time limit was from the establishment of the databases to December 31, 2019. Two reviewers independently conducted the literature screening and data extraction, and evaluated the quality according to Newcastle Ottawa scale. Stata SE 12.1 software was used for Meta-analysis.Results:Six case-control studies and 7 cohort studies met the inclusion criteria, all of which were in English. The results of Meta-analysis showed that exposure to migraine increased the risk of hemorrhagic stroke (odds ratio [ OR] 1.47, 95% confidence interval [ CI] 1.23-1.76; P<0.001). Sensitivity analysis showed that the results were robust. Subgroup analysis showed that migraine with aura ( OR 1.38, 95% CI 1.05-1.81; P=0.019), migraine without aura ( OR 1.46, 95% CI 1.19-1.80; P<0.001), male ( OR 2.10, 95% CI 1.72-2.56; P<0.001) and female ( OR 1.53, 95% CI 1.22-1.92; P<0.001) migraine could increase the risk of hemorrhagic stroke. Conclusion:Regardless of the gender of patients and presence or absence of migraine aura, migraine can significantly increase the risk of hemorrhagic stroke.

Objective:To investigate the efficacy and safety of SIMPLE regimen in the treatment of extranodal NK/T-cell lymphoma (ENKTCL).Methods:The clinical data of 11 patients with ENKTCL who were admitted to the University of Hong Kong-Shenzhen Hospital from January 2012 to January 2022 were retrospectively analyzed. The patients received 4-6 courses of SIMPLE (cisplatin, gemcitabine, ifosfamide, etoposide, dexamethasone, and pegasparaginase) regimen chemotherapy, and stage Ⅰ and Ⅱ patients who also received local radiotherapy after 2 or 3 courses of chemotherapy. Patients were evaluated for mid-treatment and end-of-treatment outcomes, and the adverse effects of patients were evaluated in each treatment cycle. The Kaplan-Meier method was used to analyze the progression-free survival (PFS) and overall survival (OS) of the 11 patients.Results:All 11 patients were nasal type, with the median age of 41 years old (26-67 years old), including 5 males and 6 females, 3 relapsed cases and 8 newly treated cases. Of the 10 patients evaluated for efficacy, 9 achieved complete remission and 1 achieved at least partial remission (efficacy was assessed based on follow-up). All 11 patients were followed up for a median time of 50 months (15-72 months) and 2 relapsed patients died due to disease progression. The expected 5-year PFS rate and OS rate of 11 patients were both 90.0%, and the expected 5-year OS rate was 100.0% and 66.6% in newly treated and relapsed patients, respectively. Common adverse effects were hematologic adverse reactions, infections, gastrointestinal symptoms, elevated transaminases, and hypofibrinogenemia, all of which were curable. There is no treatment-related death.Conclusions:The SIMPLE regimen for the treatment of ENKTCL has a high remission rate, the patients have long survival time, and the regimen is moderately well tolerated.

Objective:To establish a new model for the prediction of severe outcomes of COVID-19 patients and provide more comprehensive, accurate and timely indicators for the early identification of severe COVID-19 patients.Methods:Based on the patients’ admission detection indicators, mild or severe status of COVID-19, and dynamic changes in admission indicators (the differences between indicators of two measurements) and other input variables, XGBoost method was applied to establish a prediction model to evaluate the risk of severe outcomes of the COVID-19 patients after admission. Follow up was done for the selected patients from admission to discharge, and their outcomes were observed to evaluate the predicted results of this model.Results:In the training set of 100 COVID-19 patients, six predictors with higher scores were screened and a prediction model was established. The high-risk range of the predictor variables was calculated as: blood oxygen saturation <94 %, peripheral white blood cells count >8.0×10 9, change in systolic blood pressure <-2.5 mmHg, heart rate >90 beats/min, multiple small patchy shadows, age >30 years, and change in heart rate <12.5 beats/min. The prediction sensitivity of the model based on the training set was 61.7 %, and the missed diagnosis rate was 38.3 %. The prediction sensitivity of the model based on the test set was 75.0 %, and the missed diagnosis rate was 25.0 %. Conclusions:Compared with the traditional prediction (i.e. using indicators from the first test at admission and the critical admission conditions to assess whether patients are in mild or severe status), the new model’s prediction additionally takes into account of the baseline physiological indicators and dynamic changes of COVID-19 patients, so it can predict the risk of severe outcomes in COVID-19 patients more comprehensively and accurately to reduce the missed diagnosis of severe COVID-19.

Objective To investigate the relationship among circulating soluble major histocompatibility complex class Ⅰ-related chain A(sMICA),soluble major histocompatibility complex class Ⅰ-related chain B(sMICB),the activity of systemic lupus erythematosus(SLE)and autoantibodies.Methods A total of 156 SLE patients(SLE group)and 103 healthy volunteers(control group)who underwent physical examination in outpatient physical examination center were selected from the Qianjiang Hospital Affiliated to Chongqing University from January 2020 to January 2023.According to SLE disease activity score(SLEDAI),these SLE patients were divided into mild activity group(n=43),moderate activity group(n=69),and severe activity group(n=44).Serum levels of sMICA and sMICB,and the proportion of autoantibodies and peripheral blood NK cells were detected.Spearman or Pearson method was used to analyze the correlation among sMICA,sMICB,score,autoantibodies and peripheral blood NK cells proportion.Receiver operating characteristics(ROC)curve was used to evaluate the value of sMICA and sMICB in the diagnosis of SLE activity.Results Serum sMICA(173.65±23.92 pg/ml)and sMICB(96.35±15.74 pg/ml)levels in SLE group were higher than those in control group(32.51±6.27 pg/ml,12.03±2.47 pg/ml),while the proportion of CD3-CD56+NK cells(12.02％±2.65％)in peripheral blood was lower than that in control group(18.35％±3.71％),and the differences were statistically significant(t=58.498,53.897,-16.010,all P＜0.05).Serum sMICA and sMICB levels in severe active group were higher than those in moderately active group and mildly active group(t=8.192,12.352;19.652,23.742,all P＜0.05),and the proportion of CD3-CD56+NK cells in peripheral blood was lower than that in moderate and mild active groups(t=8.154,10.658,P＜0.05).The differences in positive rates of anti-dsDNA antibody,anti-nuclear antibody,anti-nucleosome antibody and anti-histone antibody in SLE patients with different disease activities were significant(x2=8.795,7.216,7.539,8.946,all P＜0.05).Serum sMICA and sMICB levels in SLE patients were positively correlated with SLED AI score,anti-dsDNA antibody,anti-nuclear antibody,anti-nucleosome antibody and anti-histone antibody(r=0.206～0.402,all P＜0.05),and negatively correlated with the proportion of CD3-CD56+NK cells in peripheral blood(r=-0.563,-0.427,all P＜0.05).The areas under the curve of SLE in severe active group diagnosed by sMICA and sMICB alone were 0.652 and 0.704,respectively.The area under the curve of SLE in severe active group diagnosed by sMICA and sMICB combined with SLE was 0.812,which was higher than that by the single diagnosis(Z=3.050,2.346,all P＜0.05).Conclusion The increased serum sMICA and sMICB levels in SLE patients were associated with the increased positive rate of SLE autoantibodies,the decreased proportion of NK cells in peripheral blood and the enhanced disease activity,which could be used as potential markers of SLE.

Objective:To explore the mechanism of dendritic epidermal T lymphocytes (DETCs) in promoting healing of full-thickness skin defect wound on mice by regulating the proliferation and differentiation of epidermal stem cells (ESCs) in mice.Methods:(1) Ten 8-week-old wild type (WT) male C57BL/6 mice (the same sex and kind below) were sacrificed to collect the skin of back for extracting DETCs to culture. Five WT and five 8-week-old T cell receptor (TCR) δ -/ - mice were selected and enrolled in WT control group and TCR δ -/ - control group, respectively. A full-thickness skin defect wound with diameter of 6 mm was made on both sides of spinal line on the back of mice without any treatment after injury. Another fifteen 8-week-old TCR δ -/ - mice were selected and divided into phosphate buffer solution (PBS), DETC, and insulin-like growth factor-Ⅰ(IGF-Ⅰ) groups according to the random number table (the same grouping method below), with 5 mice in each group, and the same full-thickness skin defect wound was made on each mouse. Immediately after injury, mice in PBS, DETC, and IGF-Ⅰ groups were injected subcutaneously around each wound with 10 μL sterile PBS , DETCs (cell concentration of 1×10 6/mL), and 5 mg/mL recombinant mice IGF-Ⅰ, respectively. The percentage of the residual wound area was calculated on post injury day (PID) 2, 4, 6, and 8. (2) Three 8-week-old WT mice were enrolled in WT control group and nine 8-week-old TCR δ -/ - mice were divided into TCR δ -/ - control group, PBS group, and DETC group, with 3 mice in each group. The full-thickness skin defect wound was made as in experiment (1) . On PID 3, the protein expression of IGF-Ⅰ in the epidermis tissue of wound margin was detected by chemiluminescence imaging analyzer. (3) Three 8-week-old WT mice were enrolled in WT control group and six 8-week-old TCR δ -/ - mice were divided into PBS and DETC groups, with 3 mice in each group, and the full-thickness skin defect wound was made as in experiment (1). On PID3, DETCs were extracted from the wound margin epidermis tissue to detect the percentage of DETCs expressing IGF-Ⅰ by flow cytometer. (4) The mice were taken as in experiment (2) and divided into WT control, PBS, DETC, and IGF-Ⅰ groups. A straight full-thickness skin defect incision with length of 3 cm was made in the direction of one inner ear. Mice in WT control group didn′t have any other treatment after injury, and immediately after injury, mice in PBS, DETC, and IGF-Ⅰ groups were injected subcutaneously around each wound with 10 μL sterile PBS, DETCs (cell concentration of 1×10 6/mL), and 5 mg/mL recombinant mice IGF-Ⅰ, respectively. On PID 12, epidermis tissue of wound margin was collected, and immunofluorescence staining was performed to observe the number of keratin 15 positive cells. (5) The same mice were collected, grouped, and treated as in experiment (4). On PID12, the epidermis tissue of wound margin was collected and immunofluorescence staining was performed to observe the number of keratin 10 positive cells. (6) Twenty 3-day-old WT mice (the same below) were sacrificed to collect the whole skin, which was used to extract ESCs, with 5 mice detecting one index. The ESCs were divided into DETC co-culture group and control group, which were added with 1 mL DETCs (cell concentration of 1.25×10 6/mL) and DETC medium, respectively. The percentage of 5-ethynyl-2′-deoxyuridine (EdU) positive cell on culture day (CD) 3, the percentages of CD49f + CD71 - and keratin 14 positive cells on CD 5, and the percentage of keratin 10 positive cell on CD 10 in 2 groups were detected by flow cytometer. (7) Twenty mice were taken to extract ESCs, with 5 mice detecting one index. The ESCs were divided into control group and IGF-Ⅰ group, which were added with 1 mL sterile PBS and 10 ng/mL recombinant mice IGF-Ⅰ, respectively. The percentages of EdU positive cell, CD49f + CD71 - cell, keratin10 positive cell, and keratin 14 positive cell were detected as in experiment (6). The sample in each group of experiments (6) and (7) was three. Data were statistically analyzed with analysis of variance for repeated measurement, one-way analysis of variance, and t test. Results:(1) On PID 4, 6, and 8, the percentage of residual wound area in TCR δ -/ - control group was significantly higher than that in WT control group ( t=2.78, 3.39, 3.66, P<0.05 or P<0.01). The percentage of residual wound area in DETC group and IGF-Ⅰgroup on PID 4, 6, and 8 was apparently lower than that in PBS group ( t=2.61, 3.21, 3.88, 2.84, 2.91, 2.49, P<0.05 or P<0.01). (2) On PID 3, the protein expression of IGF-Ⅰ in the epidermis tissue of wound margin of mice in TCR δ -/ - control group was significantly lower than that in WT control group ( t=17.34, P<0.01). The protein expression of IGF-Ⅰ in the epidermis tissue of wound margin of mice in DETC group was significantly higher than that in PBS group ( t=11.71, P<0.01). (3) On PID 3, the percentage of DETCs expressing IGF-Ⅰ in the epidermis tissue of wound margin of mice in PBS group was significantly lower than that in WT control group and DETC group ( t=24.95, 27.23, P<0.01). (4) On PID 12, the number of keratin 15 positive cells in the epidermis tissue of wound margin of mice in PBS group was significantly lower than that in WT control group, DETC group, and IGF-Ⅰ group ( t=17.97, 11.95, 7.63, P<0.01). (5) The number of keratin 10 positive cells in the epidermis tissue of wound margin of mice in PBS group was significantly higher than that in WT control group, DETC group, and IGF-Ⅰ group ( t=11.59, 9.51, 3.48, P<0.05 or P<0.01). (6) The percentages of EdU positive cells on CD 3, CD49f + CD71 - cells on CD 5, and keratin 14 positive cells on CD 5 in DETC co-culture group were respectively (43.5±0.6)%, (66.5±0.5)%, (69.3±1.7)%, apparently higher than (32.3±1.3)%, (56.4±0.3)%, (54.9±1.3)% in control group ( t=7.97, 17.10, 6.66, P<0.01). The percentage of keratin 10 positive cells on CD 10 in DETC co-culture group was (55.7±0.7)%, significantly lower than (67.1±1.2)% in control group ( t=8.34, P<0.01). (7) The percentages of EdU positive cells on CD 3, CD49f + CD71 - cells on CD 5, and keratin 14 positive cells on CD 5 in IGF-Ⅰ group were respectively (42.1±0.9)%, (81.1±1.3)%, (66.8±1.0)%, apparently higher than (32.4±0.7)%, (74.9±0.7)%, (52.0±1.9)% in control group ( t=8.39, 4.24, 7.25, P<0.05 or P<0.01). The percentage of keratin 10 positive cells on CD 10 in IGF-Ⅰ group was (53.5±1.1)% , significantly lower than (58.2±0.3)% in control group ( t=3.99, P<0.05). Conclusions:DETCs can promote the proliferation and anti-apoptotic potential of ESCs and inhibit their differentiation into end-stage by secreting IGF-Ⅰ, thus promoting wound healing of full-thickness skin defects in mice.

OBJECTIVETo evaluate the application of inferior vena cava filter (IVCF) in prevention of peri-operative pulmonary embolism (PE) in lower limb bone fracture patients with deep venous thrombosis (DVT).

METHODSFrom January 2003 to December 2012, 2 248 cases of lower limb and pelvic fractures with DVT were retrospectively analyzed. Before the procedure of IVCF implantation began, January 2003 to December 2007, there were 1 052 cases of acute trauma patients with DVT were classified as the group of early none-IVCF. The IVCF implantation was began since January 2008. From that time to December 2012, 712 cases of bone fractures with DVT received filter implantation, which were classified as IVCF group. The other 484 patients who had not undergone filter deployment were divided as group of late none-IVCF. The baseline conditions of the three groups were significantly different in addition to the ages between group of early none-IVCF and IVCF group. The incidences of PE and mortality of PE in each group were recorded and analyzed by χ(2) test.

RESULTSThere were totally 31 cases of symptomatic PE, among which 12 cases died. Totally 712 filters were deployed successfully without any major complications. The incidences of symptomatic PE were 0.14% (1/712), 2.19% (23/1 052) and 1.45% (7/484) in IVCF group, group of early none-IVCF and group of late none-IVCF, respectively. The mortality of PE were 0 (0/712), 0.86% (9/1 052) and 0.62% (3/484) in these groups. The incidence of symptomatic PE in IVCF group was significantly different from that in the group of early and late none-IVCF (χ(2) = 11.762, P = 0.001; χ(2) = 7.395, P = 0.007, respectively). The mortality of IVCF group was also significantly lower compared with the other two groups (χ(2) = 6.122, P = 0.013; χ(2) = 4.424, P = 0.035, respectively).

CONCLUSIONIVCF implantation effectively prevents symptomatic and fatal PE of patients of lower limb and pelvic fractures with DVT in the peri-operative period.

Adult ; Aged ; Female ; Humans ; Leg Injuries ; complications ; Lower Extremity ; Male ; Middle Aged ; Pulmonary Embolism ; etiology ; prevention & control ; Retrospective Studies ; Vena Cava Filters ; Venous Thrombosis ; complications

Coronavirus disease 2019 (COVID-19) outbroke in Wuhan, China in December 2019 and the severe acute respiratory syndrome (SARS) outbroke in Guangzhou, China in 2003 were caused by highly pathogenic coronaviruses with high homology. Since the 2019 novel coronavirus is highly contagious and spreads rapidly. It has caused negative social effects and massive economic loss globaly. Currently there is no vaccine or effective drugs. Pulmonary fibrosis is a pulmonary disease with progressive fibrosis, which is the main factor leading to pulmonary dysfunction and declined quality of life in SARS survivors after recovery. Extensive epidemiological, viral immunological and current clinical evidences support the possibility that pulmonary fibrosis may be one of the major complications in COVID-19 patients. At present there is no report on the mechanism by which COVID-19 induces pulmonary fibrosis.With the existing theoretical basis, this article focuses on discussing the possible mechanism of COVID-19 sustained lung damaging, the key role of abnormal immune mechanism in the initiation and promotion of pulmonary fibrosis, and the corresponding therapeutic measures.