BACKGROUND:Compared with traditional two-dimensional culture,three-dimensional microtissue culture can show greater advantages.However,more favorable cultivation methods in three-dimensional culture still need to be further explored. OBJECTIVE:To evaluate the cell behavior of microtissue and its ability to promote cartilage formation under two three-dimensional culture methods. METHODS:Cartilage-derived microcarriers were prepared by chemical decellularization and tissue crushing.DNA quantification and nuclear staining were used to verify the success of decellularization,and histological staining was used to observe the matrix retention before and after decellularization.The microcarriers were characterized by scanning electron microscopy and CCK-8 assay.Cartilage-derived microtissues were constructed by combining cartilage-derived microcarriers with human adipose mesenchymal stem cells through three-dimensional static culture and three-dimensional dynamic culture methods.The cell viability and chondrogenic ability of the two groups of microtissues were detected by scanning electron microscopy,live and dead staining,and RT-qPCR. RESULTS AND CONCLUSION:(1)Cartilage-derived microcarriers were successfully prepared.Compared with before decellularization,the DNA content significantly decreased after decellularization(P<0.001).Scanning electron microscope observation showed that the surface of the microcarrier was surrounded by collagen,maintaining the characteristics of the natural extracellular matrix of cartilage cells.CCK-8 assay indicated that microcarriers had no cytotoxicity and could promote cell proliferation.(2)Scanning electron microscopy and live and dead staining results showed that compared with the three-dimensional static group,the three-dimensional dynamic group had a more extended morphology of microtissue cells,and extensive connections between cells and cells,between cells and matrix,and between matrix.(3)The results of RT-qPCR showed that the expressions of SOX9,proteoglycan,and type Ⅱ collagen in microtissues of both groups were increased at 7 or 14 days.The relative expression levels of each gene in the three-dimensional dynamic group were significantly higher than those in the three-dimensional static group at 14 days(P<0.05).At 21 days,the three-dimensional static group had significantly higher gene expression compared with the three-diomensional dynamic group(P<0.001).(4)The results showed that compared with three-dimensional static culture microtissue,three-dimensional dynamic culture microtissue could achieve higher expression of chondrogen-related genes in a shorter time,showing better cell viability and chondrogenic ability.

Objective:To evaluate the effect of transcutaneous electrical acupoint stimulation (TEAS) on function of lung ventilation in the patients undergoing laparoscopic cholecystectomy using electrical impedance tomography (EIT).Methods:Sixty American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients of both sexes, aged 18-64 yr, with body mass index of 18.5-24.9 kg/m 2, undergoing elective laparoscopic cholecystectomy, were divided into 2 groups ( n=30 each) using a random number table method: control group (CON group) and TEAS group.EIT was carried out after entering the operating room.In group TEAS, bilateral Feishu (BL13) and Chize (LU5) acupoints were stimulated, while stimulation electrodes were only placed without electricity in group CON.The percentages of area in center of ventilation and tidal volume in the dependent areas were determined on admission to the operating room (T 1), at 5 min after tracheal intubation (T 2), at 5 min after CO 2 pneumoperitoneum (T 3), at 5 min after CO 2 was exhausted (T 4), and at 5 and 30 min after removal of the tracheal tube (T 5, 6). Radial artery blood samples were collected for blood gas analysis at T 1, T 3 and T 6, and the oxygenation index (OI) and alveolar-arterial oxygen difference (A-aDO 2) were calculated.Blood samples were taken from the peripheral vein at T 1, T 3 and T 6 for determination of the serum concentrations of Clara cell secretion protein (CC16), interleukin (IL)-6 and tumor necrosis factor (TNF)-α. Results:Compared with the baseline at T 1, the percentages of area in center of ventilation and tidal volume in dependent areas at T 2-6 and OI and A-aDO 2 at T 6 were significantly decreased, and the serum concentrations of TNF-α, IL-6 and CC16 were increased at T 3 and T 6 in two groups ( P<0.05). Compared with group CON, the percentage of tidal volume in dependent areas at T 5, 6 and OI at T 6 were significantly increased, and A-aDO 2 and serum concentrations of TNF-α, IL-6 and CC16 were decreased at T 6 in group TEAS ( P<0.05). Conclusions:TEAS can improve the function of lung ventilation in the patients undergoing laparoscopic cholecystectomy.

BACKGROUND: microRNAs play an important role in the development and biological phenotype of lung cancer. The present study was to investigate miR-367-3p level in non-small cell lung cancer (NSCLC) patients and its biological function of NSCLC cells. METHODS: Twenty-two patients with NSCLC (13 cases of adenocarcinoma and 9 cases of squamous carcinoma) admitted to our hospital and treated by surgery were included. During the operation, cancer tissue, paracancerous tissue and 5 mL peripheral blood were collected. Meanwhile, 22 healthy controls were selected and 5 mL peripheral blood was taken. Real-time PCR was applied to detected the expression of miR-367-3p in cancer tissues, peripheral blood of patients with NSCLC and healthy controls. miR-367-3p was detected in lung cancer cell lines (A549) and normal bronchial epithelial cells (BEAS-2B). The proliferation and invasion ability of A549 cells before and after infection were detected by MTT and Transwell assay after transfection with exogenous miR-367-3p. The downstream target gene of miR-367-3p was analyzed by bioinformatics. Zinc finger E-box binding homeobox 2 (ZEB2) was detected by Real-time PCR and Western blot. RESULTS: miR-367-3p in cancer tissues of 22 NSCLC patients was lower than corresponding normal tissues (P<0.05), and the serum miR-367-3p level in healthy subjects was higher than NSCLC subjects (P<0.05). The area under the receiver operating characteristic (ROC) curve of NSCLC was 0.95 (95%CI: 0.89-1.00) and 0.85 (95%CI: 0.74-0.97) respectively; The proliferation and migration ability of lung cancer cell line A549 transfected with exogenous miR-367-3p decreased significantly (P<0.05); Bioinformatics predicted that the downstream target of miR-367-3p was ZEB2 and up-regulating miR-367-3p expression, ZEB2 gene was decreased (P<0.05). The Cancer Genome Atlas (TCGA) data analysis showed that there was no significant difference in overall survival (OS) and disease free survival (DFS) between ZEB2 high expression group and low expression group (P>0.05). ZEB2 expression was positively correlated with infiltration of B lymphocytes (r=0.32, P<005), CD8⁺ T cells (r=0.44, P<005), CD4⁺ T cells (r=0.46, P<005), macrophages (r=0.65, P<005), neutrophils (r=0.73, P<005) and dendritic cells (r=0.71, P<005) in NSCLC. CONCLUSIONS The expression of miR-367-3p is down regulated in NSCLC patients and participates in the biological process of proliferation and invasion of NSCLC by targeting ZEB2 gene.
Humans ; Carcinoma, Non-Small-Cell Lung/genetics* ; Lung Neoplasms/genetics* ; Cell Proliferation ; MicroRNAs/genetics* ; A549 Cells ; Zinc Finger E-box Binding Homeobox 2/genetics*

Objective:To investigate the clinical efficacy of intramedullary nailing combined with bone cement in repair of segmental bone defects after tumor resection.Methods:A retrospective analysis was conducted of the 5 patients with malignant bone tumor who had been treated at Department of Orthopaedics, Qiannan People's Hospital from April 2018 to September 2019 for remaining segmental bone defects following limb salvage surgery. They were 4 males and one female, aged from 11 to 55 years (average, 35.4 years). Their defects ranged from 6 to 21 cm (average, 12.3 cm) after tumor resection. By the Karnofsky performance score (KPS) for long-term quality of life, all of them scored less than 50 points. Of them, 3 were treated by interlocking intramedullary nails and bone cement filling, and 2 by elastic intramedullary nails and bone cement filling. In the 2 cases with defects of 21 cm and 13 cm, the fixation was assisted by a plate and an external fixator. Defect length after resection, operation time and intraoperative bleeding were recorded; the efficacy was evaluated by the Enneking functional evaluation of reconstructive procedures after surgical treatment of tumors of the musculoskeletal system, visual analogue scale (VAS), and KPS.Results:All the 5 patients had uneventful surgery, with operation time ranging from 112 to 225 min (average, 154.2 min), intraoperative bleeding from 300 to 500 mL (average, 382 mL), and defect length after resection from 6 to 21 cm. The 5 patients were followed up for 6 to 28 months. Of them, 2 died of disease progression 6 and 7 months after surgery, respectively. According to the Enneking's evaluation, one patient scored 28 points, 2 patients 23 points and 2 patients 21 points, giving a high degree of satisfaction. Their VAS scores 6 months after surgery ranged from 1 to 6, averaging 3.6; their postoperative KPS scores ranged from 60 to 80, averaging 72.Conclusion:In repair of segmental bone defects after tumor resection, intramedullary nailing combined with bone cement filling can relieve pain of patients and lead to satisfactory short-term curative efficacy.

Objective:To analyze the clinical efficacy of scleral buckling surgery for rhegmatogenous retinal detachment (RRD) of 376 patients.Methods:A retrospective analysis was performed about 376 patients (391 eyes) who underwent scleral buckling surgery in Chengdu Aidi Eye Hospital from January 2018 to December 2019. There were 214 males (224 eyes) and 162 females (167 eyes). There were 15 binocular cases and 361 monocular cases. The average age was 37.16±16.36 years. The average course of disease was 3 months. There were 1 to more than 10 retina holes for all patients. Retinal breaks occur in all quadrants and at ora serrata. The preoperative average BCVA was 0.27 and the postoperative average BCVA was 0.41. Retinal detachment ranges was observed in 268 eyes in 1 quadrant, 97 eyes in 2-3 quadrants, 26 eyes in total, and 231 eyes with macular involvement. There were 376 eyes treated with scleral buckling, 9 eyes treated with scleral buckling combined with scleral encircling, 6 eyes treated with scleral encircling. The average follow-up time was 5 months. Postoperative follow-up was conducted to observe retinal reduction, BCVA, complications and patient compliance.Results:After the first operation, retinal reattachmnents were successfully achieved in 375 eyes (95.91%); 16 eyes (4.09%) failed in retinal reattachmnents. Eight eyes were treated with scleral buckling again, 5 eyes were treated with vitrectomy silicone oil filling, and 3 eyes were treated with air injection. After the second operation, retinal reattachmnents were ultimately achieved in 16 eyes (100.00%). The average BCVA after operation was 0.15. Postoperative intraocular pressure increased by 45 eyes (11.51%). The intraocular pressure increased from the next day to 3 days after operation. The intraocular pressure was completely controlled 1-3 days after the treatment of topical medication and 20% mannitol. Vitreous and subretinal hemorrhage in 1 eye caused by drainage of the subscleral liquid. There was no cases with postoperative infection.Conclusion:The retinal reattachment rate is 95.91% in 376 patients with RRD treated by scleral buckling surgery, and the visual acuity has significantly improved.

Objective:To evaluate the accuracy of ultrasound-measured pulsatility index (PI) of central retinal arteries in diagnosing low cerebral perfusion pressure (CPP) in the patients with traumatic brain injury.Methods:Fifty-five patients who were admitted to the surgical intensive care unit due to traumatic brain injury with continuous intracranial pressure monitoring were selected.At 72 h after surgery, continuous incracranial pressure and mean arterial pressure were recorded, and peak systolic velocity and end diastolic velocity of central retinal arteries were measured by ultrasound.The invasive CPP and PI were calculated.The correlation between PI and CPP was analyzed.The accuracy of central retinal pulse index in diagnosing low CPP was evaluated by the receiver operating characteristic curve.Results:PI was negatively correlated with CPP ( r=-0.655, P<0.01). The area under the receiver operating characteristic curve of PI in diagnosing low CPP was 0.863 (95% confidence interval 0.761-0.965), and the threshold value was 0.97, sensitivity 92.3%, and specificity 66.7%. Conclusion:Ultrasound-measured PI is more accurate in diagnosing low CPP in the patients with traumatic brain injury.

Objective : To learn the incidence trend of gestational diabetes mellitus (GDM) among women of childbearing age in Hangzhou from 2009 to 2018, providing evidence for GDM prevention and control. Methods: The information of GDM cases aged 15 to 49 years in Hangzhou from 2009 to 2018 was extracted from Zhejiang non-communicable diseases surveillance system. The crude and age-standardized (by national standard population in 2000) incidence rate of GDM were calculated, the annual change percentage (APC) and the average APC (AAPC) of GDM incidence rate were calculated to understand the trend of incidence in different age groups and areas. Results: A total of 45 519 cases of GDM were reported in Hangzhou from 2009 to 2018. The crude and age-standardized incidence rates were 248.04/100 000 and 158.52/100 000, respectively, both showed upward trends with 59.7% and 56.4% in AAPC (both P<0.05). The APCs of crude and age-standardized incidence rates of GDM were 139.76% and 134.60% from 2009 to 2012, 30.35% and 27.65% from 2012 to 2018, which all had increasing trends (all P<0.05). There were upward trends in GDM incidence in each age groups of 15-24, 25-29, 30-34, 35-39 and 40-49 years old (all P<0.05). The AAPC of GDM incidence in 40-49 year-old women was 65.3% and ranked the first among all the age groups. The age-standardized incidence rate of GDM increased more in rural areas (AAPC=66.0%) than in urban areas (AAPC=53.4%). Conclusion A rising trend in GDM incidence among women aged 15-49 years in Hangzhou has been found in recent ten years, especially in women aged 40-49 years and lived in rural areas.

Objective: To evaluate the accuracy of ultrasound-determined end-diastolic velocity (EDV) of central retinal artery (CRA) in diagnosing postoperative low cerebral perfusion pressure (CPP) in the patients with craniocerebral trauma. Methods: Forty-nine patients of both sexes with brain injury, aged 18-64 yr, with body mass index of 18.5-23.9 kg/m², were enrolled.The peak systolic velocity and EDV of CRA were determined using ultrasound at 1 day after operation.Mean arterial pressure and intracranial pressure were recorded, and CPP was calculated (CPP=mean arterial pressure-intracranial pressure). Results: EDV was positively correlated with CPP (r=0.746, P<0.01), and peak systolic velocity was not correlated with CPP (P>0.05). The area under the receiver operating characteristic curve for EDV in diagnosing low CPP was 0.938 (95% confidence interval 0.871-1.000), and the critical value was 3.205 (sensitivity 94.4%, specificity 76.9%). Conclusion Ultrasound-determined EDV of central retinal artery can accurately diagnose postoperative low CPP in the patients with craniocerebral trauma.

Objective To analyze the relationship between onset of myocardial infarction(MI)to bivalent antibiotics therapy and outcome in elderly female MI patients.Methods One hundred and fourteen elderly MI patients admitted to our hospital from January 2016to January 2018were divided intoΔT≤188min group(n=57)andΔT＞188min group(n=57)according to the onset of MI to bivalent antibiotics therapy.The major adverse cardiac events(MACE),arrhythmia,angina pectoris,heart failure and death were compared between the two groups.Results MACE occurred in 44out of the 144MI patients included in this study.Of the 44MI patients,17died,6 were diagnosed with arrhythmia,16were diagnosed with angina pectoris,5were diagnosed with heart failure.The incidence of arrhythmia,heart failure and the mortality were significantly higher while the incidence of angina pectoris was significantly lower inΔT≤188min group than inΔT＞ 188min group(7.0％vs 3.5％,7.0％vs 1.8％,15.8％vs 14.0％,7.0％vs 21.1％,P＜0.05).Age,diabetes,ΔT,hyperlipidemia,hypertension and coronary artery disease were the independent influencing factors for the poor outcome in MI patients(P＜0.05,P＜0.01).Conclusion TheΔT should be shortened,the MI patients should be timely treated,the criminal blood vessels should be opened,and the ischemic myocardium should be saved in order to prevent the occurrence of severe cardiovascular events and reduce the mortality in elderly MI patients.

Background and purpose: Cervical conization is a common operation to treat precancerous tissues performed under non-intubated anesthesia. As common opioid analgesics have side effects of inhibiting respiration and circulation, other kinds of analgesic drugs should be coordinated to improve the anesthetic effect, without interfering the respiration and circulation. This study aimed to evaluate the effects of dezocine or flurbiprofen combined with propofolremifentanil in cervical precancerosis conization. Methods: Sixty patients who underwent cervical conization were equally randomized into dezocine group (group D), flurbiprofen group (group F) and 0.9％ natural saline (group N) with 20 patients in each group, and received dezocine 0.1 mg/kg, flurbiprofen 1 mg/kg or 0.9％ natural saline in 5 mL respectively before anesthesia induction. During the anesthesia induction, the targeted control infusion of remifentanil in effect concentration was set at 1.5 ng/mL, and the plasma concentration of propofol was set at 2 μg/mL. Heart rate (HR), respiratory rate (RR), surplus pulse O2 (SPO2) and mean arterial pressure (MAP), MAP were monitored before the anesthesia induction (T0) and after (T1), at the start of cervical conization (T2), and at the end of operation (T3). The incidence of respiratory depression and body movements during surgery were observed. The satisfaction degree of the surgeon to the opening status of cervix was evaluated. The post-operative recovery time, visual analogue scale (VAS) scores, nausea and vomiting in the following 12 hours were also recorded. Results: The HR, RR, SPO2 and MAP in three groups did not have any significant change (P＞0.05) at T0, T1 and T3. At T2 the HR and MAP decreased significantly in group D and group F compared with group N (P＜0.05), and there was no significant difference between group D and group F (P＞0.05). The surgical satisfaction degree of "Good" in group D was 80％, significantly higher than that in group N (30％) and group F (50％), indicating a better cervix opening in group D. The recovery time in three groups had no significant difference, and the VAS scores in group D and group F were lower than those in group N (P＜0.05) after operation, and patients did not have nausea or vomiting in the following 12 hours. Conclusion: Both the dezocine and flurbiprofen could improve the anesthetic effect in cervical conization and post-operative comfort, with less respiratory or circulation depression. Dezocine showed better improvement than flurbiprofen in cervix opening and the inhibition of stress response and body movements during surgery.