Objective: To observe the regulatory effects of miRNA-31 (miR-31) on LATS2 and cardiomyocyte hypertrophy via down-regulating miR-31 expression in rat's cardiomyocytesin vitro. Methods: Rat's cardiomyocytes were isolated and cultured for 10 daysin vitro, according to different intervention methods, the cells were divided into 4 groups:①Blank control group,②AngII intervention group,③Lentivirus with miR-31 inhibitor infection group,④Negative lentivirus infection group. On day-8, gene expressions of MiR-31, LATS2, cardiac hypertrophy ANP and β-MHC were examined by qRT-PCR; on day-10, cell morphology was observed by fluorescence staining. LATS2 protein expression was examined by Western blot analysis. Dual luciferase reporter plasmids were transfected into 293T cells, then luciferase activity was detected to identify the targeting effect of miR-31 on LATS2. Results: Compared with Blank control group, AngII intervention group showed increased gene expressions of miR31, cardiac hypertrophy ANP and β-MHC,P<0.05, enlarged cardiomyocyte surface,P<0.05; while decreased gene and proteinexpressions of LATS2,P<0.05. Compared with AngII intervention group, Lentivirus with miR-31 inhibitor infection group had down-regulated expressions of miR31, cardiac hypertrophy ANP and β-MHC,P<0.05, reduced cardiomyocyte surface, P<0.05; while slightly increased LATS2 gene expression and obviously increased protein expression,P<0.05. Dual luciferase reporter assay presented that relative luciferase activity of TRAF6-3' UTR+miR-146b was significantly decreased than TRAF6-3' UTR+miR-NC,P<0.01 and relative luciferase activity of LATS2-3' UTR+ miR-31 was signiifcantly reduced than LATS2-3' UTR-NC+miR-31,P<0.01. Conclusion: Cardiomyocytes hypertrophy could be reversed at certain degree by down-regulating miR-31; the targeting effect of miR-31 on LATS2 was involved in cardiomyocyte hypertrophyregulation.

Objective To investigate the prevalence and risk factors of chronic kidney disease (CKD) in Chengdu urban population and the prevalence of CKD in risk population.Methods Questionnaire (anamnesis,smoking,drink) of risk factors of CKD and somatoscopy (blood pressure,body height and body weight) were caried out in railman of Chengdu urban.Their blood and urine indicators (blood sugar,blood lipid,blood uric acid,blood creatinine,uromicroprotein/creatinine ratio,routine urine examination,etc) were measured.The prevalence and risk factors of CKD in Chengdu urban population and the prevalence of CKD in risk population were elucidated.Results Eligible data of 5326 subjects were enrolled in the study.After the adjustment of age and gender component,the prevalence of albuminuria was 11.54％,reduced eGFR was 5.54％,hematuria was 3.87％,and CKD was 18.32％; the recognition was 1.93％.In addition,the prevalence of albuminuria was respectively 23.79％,28.00％,14.08％; prevalence of reduced eGFR was respectively 4.76％,4.53％,3.26％; prevalence of hematuria was respectively 2.94％,3.20％,2.37％ in 3098 people with hypertension,diabetes or hyperlipaemia.Independent risk factors of albuminuria were female,hypertension,diabetes,hyperlipemia and high BMI.Independent risk factors of reduced eGFR were female,age,hyperuricemia and hypertension.Drink was negatively correlated with reduced eGFR.Independent risk factors of hematuria were female and age.Conclusions The prevalence of CKD is quite high and the recognition rate is low in the Chengdu urban populaton.Risk factors of CKD are age,female,diabetes,hypertension,hyperlipemia,hyperuricemia and high BMI.Control of the development of metabolic disease can reduce the CKD.

BACKGROUND: It has been believed mesenchymal stem cells (MSCs) play a role in treatment through paracrine mechanism. Various side effects such as embolism, tumorigenesis and immunological reaction caused by direct injection of MSCs can be avoided by extracting MSC lysate. However, there is a larger difference in current collection methods and standards of MSC lysate. OBJECTIVE: To compare repeated freeze-thaw and ultrasonication for the collection of lysate of MSCs. METHODS: Adipose-derived mesenchymal stem cells (ADMSCs) were isolated from the abdominal subcutaneous fat of healthy individuals, and purified with adherence screening method, followed by in vitro amplification using fetal bovine serum medium. The common surface makers of these cells were tested by flow cytometry (1×109, 2×109, 4×109/L). Repeated freeze-thaw and ultrasonication were employed for cell cytoclasis at three different densities respectively in saline and double distilled water, and a comprehensive comparison was performed on cytoclasis rate and the content of protein in cell lysate between the two methods. RESULTS AND CONCLUSION: (1) ADMSCs obtained from in vitro isolated human adipose tissue grew in a swirl or radial pattern with a homogenous size and neat arrangement. CD44, CD90, CD105 and other commonly used surface markers were highly expressed. (2) The study for optimization of lysate collection revealed that the higher cell density implicated a longer time for cell wall disruption and cytoclasis, as well as significantly increased cytoclasis rate. (3) BCA protein assay showed that the highest content of protein was obtained in saline solvent using ultrasonication method. Comprehensive analysis on the results leads to a conclusion that ultrasonication method with saline as the solvent is the optimized method for extraction of ADMSCs lysate, and the cell concentration of less than 4×109/L is recommended.

Objective To investigate the clinical and molecular genetic changes in a Chinese family with oculopha?ryngeal muscular dystrophy(OPMD). Methods We collected the clinical data of the familial members and blood sam?ples from all available 16 familial members, including the proband. The samples were analyzed using modified poly?merase chain reaction amplification and direct sequence analysis. Results Male OPMD patients initially presented with ptosis, followed by pronunciation difficulty, dysphagia and limb weakness whereas female OPMD patients initially pre?sented with swallowing difficulty. Genetic test revealed the abnormal expansions of the GCG trinucleotide repeat from GCG6 to GCG10 in PABPN1 gene in 10 familial members. Conclusions The genetic test and prenatal diagnosis is the key for the prevention treatment of oculopharyngeal muscular dystrophy. The ptosis of eyelid may be the initial symptom for the male patients of oculopharyngeal muscular dystrophy with (GCG)10 mutation.

Rectal cancer with simultaneous liver metastasis is very common clinically. R0 surgical resection both for the original and metastatic tumor can achieve much better long-term oncological results. The operation types include traditional open procedures for both rectal cancer and liver metastatic resection; combination of laparoscopic resection of the rectal cancer and open procedure resection of the liver metastatic lesion; traditional laparoscopic-assisted rectal and liver metastatic tumor resection with small abdominal incision and total laparoscopic natural orifice specimen extraction surgery(NOSES) without abdominal incision. Due to the complexity of rectal anatomy and treatment strategy, leading to the difference from colon cancer with liver metastasis, and due to the effect of laparoscopic treatment, especially the 3D laparoscopy, patient selection for simultaneous resection should be well planned and individualized by surgeons based on conditions of themselves and patients.

Objective:This study aims to analyze the counts (per kilogram of body weight) or percentages of transplanted lymphocyte subgroups in children with non-infectious pulmonary complications (NIPC) and air-leak syndrome (ALS) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and explore its significance in the progression of lung complications after transplantation.Methods:The patients with NIPC and ALS after allo-HSCT from January 2013 to December 2019 in Hebei Yanda Ludaopei Hospital were retrospectively studied and the influencing factors in the progress of NIPC after HSCT were statistically analyzed.Results:Of the 2026 children who received HSCT treatment, 59 patients (34 males and 25 females) developed NIPC, the probability was 2.9% (59/2 026), and the probability of combined ALS was 1.4% (28/206). The differences in the comparison between NIPC progressed to ALS group (ALS group) and failed to progress to ALS group (non-ALS group) in the patient′s age( P=0.028), disease condition before transplantation( P=0.022), NIPC onset time( P=0.004) were significant. The P values of the percentage of NKT-like cells in the bone marrow ( P=0.008) or peripheral stem cells ( P=0.003) accounted for the lymphocytes. CD4+CD25+dim cells in bone marrow ( P=0.029) or peripheral stem cells ( P=0.036) accounted for the CD4+lymphocytes and the ratio of CD4/CD8 in bone marrow( P=0.004) or peripheral stem cells ( P=0.020) were less than 0.05, which meant the differences in patients′ refusion cells were significant. In the binary logistic regression model, the percentage of bone marrow NKT-like cells to lymphocytes, the ratio of bone marrow CD4+/CD8+and the percentage of peripheral stem NK cells to lymphocytes were important risk factors for the progression of NIPC to ALS. The rest factors were excluded from the model (AUC=0.918, P<0.05). Conclusion:During allo-HSCT transplantation, a high proportion of NKT-like cell and NK cell levels, and a high CD4+/CD8+ratio in the infusion of donors with high immune tolerance have an important correlation with the progression of the NIPC.

OBJECTIVETo introduce the use of a self-made specimen protective sleeve in laparoscopic resection for upper or mid rectal cancer and sigmoid colon cancer with transrectal specimen extraction surgery and the improvement of implantation method, so as to avoid and reduce bacterial contamination and tumor cell dissemination in abdominal cavity.

METHODSDuring June 2015 and May 2017, 48 cases of high located rectal or sigmoid colon cancer were operated laparoscopically with natural orifices specimen extraction surgery (NOSES) using a self-made specimen protecting sleeve. Operation indication: (1) Rectum and sigmoid colon cancer with the distance of more than 6 cm from tumor inferior margin to dentate line. (2) The maximum diameter of intestine together with mesangial and tumor <7 cm by intraoperative judgment. (3) No anal and distal rectal surgery, no anorectal stenosis or lack of expansion capacity caused by trauma. (4) No ulcerative colitis, Crohn's disease or radiation proctitis. After transecting the rectum, the specimen protective sleeve was inserted through the right lower 12 mm main Trocar (This sleeve was tailored from the laparoscopic protective sleeve produced by China 3L Corporation, which was intercepted with 25-35 cm from one end of the sleeve according to the length of distal rectal retention. One end was ligated and the other was open with a ligature band. About 5 ml paraffin oil was used to rinse and lubricate during the operation). The rectal stump retained 7-8 cm in abdominal cavity. The transanal ligation part of the protective sleeve was cut off, then the stapler nail seat was inserted and specimen was pull out through the sleeve and rectum.

RESULTSThere were 30 males and 18 females. The average age was (64.5±14.1) years, the BMI was (25.4±3.9) kg/m, the tumor diameter was (3.3±1.1) cm, the maximum diameter of specimen was (5.4±1.5) cm and the length of specimen was (18.6±4.3) cm. Among these 48 cases, specimens of 36 patients were pulled out through inside of the sleeve easily, while specimens of 12 patients were quite difficult with resistance. Of 12 cases, 7 needed the help of transverse forceps, 4 needed to make 1 cm incision in pull-through bowel and insert a suction to decrease the volume of large specimens with gathering of gas and fluid, and 1 received small abdominal incision to remove specimen and perform intestinal reconstruction due to big specimen (the diameter of tumor and mesentery was 7.5 cm). Specimen tears of 6 patients didn't result in dissemination thanks to the specimen protecting sleeve. The operation time was (113.2±76.1) min, the bleeding amount was (38.5±17.3) ml, the time to first oral intake was (47.9±4.4) h, and the postoperative hospitalization length was (8.5±1.7) d. Anastomotic leakage occurred in 1 case (2.1%). No intra-abdominal and trocar infection, and obstruction were found.

CONCLUSIONThe use of protective sleeve and the improvement of the method of intraperitoneal implantation can effectively reduce the abdominal contamination during the specimen extraction. It can be applied to big specimens as well.

Objective:To identify the factors associated with successful transrectal specimen extraction after laparoscopic rectal cancer resection.Methods:A retrospective case-control study was conducted. Clinical data of rectal cancer patients who did or did not successfully undergo transrectal specimen extraction in Shanghai East Hospital between January 2017 and December 2017 were retrieved through the rectal cancer database of Shanghai East Hospital. Case inclusion criteria: (1) tumor size ≤7 cm by pelvic MRI; (2) body mass index (BMI)≤ 30 kg/m 2; (3) no history of neoadjuvant chemoradiotherapy; (4) no anal stenosis. Clinical data including age, gender, BMI, tumor obstruction, distance from tumor to anal verge, history of abdominal operation, maximal diameter of tumor and width of mesorectum in the anteroposterior dimension measured by pelvic MRI, etc. were collected. The χ 2 test was used to perform univariate analysis. Multivariate logistic regression was used to identify factors affecting transrectal specimen extraction. Results:A total of 208 patients were included in the analysis. Of 208 patients, 132 were men and 76 were women; mean age was (63±11) years old and median tumor size was 3.8 (IQR, 3.0 to 5.0) cm. Sixty-six (31.7%) patients completed transrectal specimen extraction successfully. Univariate analysis showed that patients who completed transrectal specimen extraction were more likely to have a lower BMI (χ 2=7.420, P=0.006), be free from malignant obstruction (χ 2=8.972, P=0.003), have a shorter distance from tumor to the anal verge (<5.0 cm) (χ 2=14.960, P<0.001), a smaller tumor size (≤5.0 cm) (χ 2=18.495, P<0.001) and a thinner mesorectum in the anteroposterior dimension (≤6.0 cm) (χ 2=34.612, P<0.001) than those who failed to perform transrectal specimen extraction. Gender, age or history of abdominal operation were not associated with the successful extraction (all P>0.05). Multivariate analysis revealed that BMI ≤25.0 kg/m 2 (OR=2.32, 95% CI: 1.06 to 5.06, P=0.034), free from malignant obstruction (OR=3.01, 95% CI: 1.82 to 6.69, P<0.001), the distance from tumor to the anal verge <5.0 cm (OR=3.73, 95% CI: 1.22 to 11.43, P=0.021), tumor size ≤ 5.0 cm (OR=4.43, 95% CI: 1.39 to 14.09, P=0.012), and the anteroposterior width of mesorectum ≤ 6.0 cm (OR=4.30, 95% CI: 2.02 to 9.18, P<0.001) were independent protective factors for successful transrectal specimen extraction. Conclusion:Preoperative assessment of BMI, malignant obstruction, distance from tumor to the anal verge, tumor size and anteroposterior width of mesorectum is beneficial to choose appropriate patients with rectal cancer to undergo transrectal specimen extraction.

Objective:To explore and screen the immunophenotypic characteristics of acute promyelocytic leukemia (APL) by multiparameter flow cytometry (MFC).Methods:A retrospective and descriptive study. A total of 130 acute myeloid leukemia (AML) patients who registrated in Hebei Yanda Lu Daopei Hospital were studied, among which there were 44 classical APL (cAPL), 24 microgranular variant of APL (APLv) and 62 non-APL patients (including NPM1 mut AML and AML with KMT2A rearrangement). MFC immunotyping was used to analyze and compare the median expression intensity (MEI) of side scatter (SSC), along with the ratio of the MEI on leukemic cells with those on lymphocytes (T/L MEIR), the median fluorescence intensity (MDFI) of CD34, myeloperoxidase (MPO), CD64 and CD9 on leukemic cells, as well as the ratios of these MDFIs on leukemic cells with those on lymphocytes (T/L MDFIR). Receiver operating characteristic (ROC) curve was drawn to evaluate the diagnostic efficiency of the multiparameters model for distinguishing cAPL and non-APL, APLv and non-APL. Results:The MEI and T/L MEIR of SSC in the cAPL group were higher than those in the APLv and non-APL groups ( P<0.05), and these two parameters in APLv group were higher than those in the non-APL group, respectively ( P<0.05). The MDFIs of CD34 in cAPL and APLv groups were higher than those in the non-APL group ( P<0.05), and the T/L MDFIR of CD34 was higher in APLv group than non-APL group ( P<0.05). The MDFIs of MPO and CD9, as well as the T/L MDFIRs in cAPL and APLv groups were both higher than those in the non-APL group, respectively ( P<0.05). The MDFI and T/L MDFIR of CD64 in the cAPL group were higher than those in non-APL group, respectively ( P<0.05). ROC curve results showed that the area under the curve (AUC) of MEI of SSC, the MDFI of CD64 and CD9, as well as the T/L MEIR of SSC and T/L MDFIR of CD9 were 0.932, 0.816, 0.893, 0.960 and 0.894 for diagnosing cAPL, respectively, and the AUC of these parameters were 0.725, 0.737, 0.791, 0.729 and 0.736 for diagnosis APLv, respectively ( P<0.05). Conclusion:MFC method can analyze and screen the immunophenotypic characteristics of APL for differential diagnosis of cAPL, APLv and non-APL patients.