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@#Objective To study the effect of growth hormone(GH)with progesterone-primed ovarian stimulation(PPOS)protocol on in vitro fertilization(IVF)outcomes among women with low prognosis.Methods This is a retrospective cohort study using propensity score matching(PSM)analysis.Women commencing their IVF between January2017 to December2021,with and without GH co-treatment,were reviewed.Results After PSM,76 pairs of women with low prognosis were included into analysis.Paired testing showed there is a statistical increase in transferable embryo(P<0.001)in the GH co-treatment group comparing with control group.No significant difference showed in retrieved oocyte number and metaphaseⅡ(MⅡ)oocyte rate,two-pronuclear(2PN)zygote rate on day1,high-quality embryo rate,or clinical pregnancy rate,neither in gonadotropin(Gn)requirement,duration or peak estradiol.Subgroup analysis results showed transferrable embryo rate on day3 rising among decreased ovarian reserve women(P=0.010).For women aged below 40,MⅡoocyte rate(P=0.010)and transferrable embryo rate on day3(P<0.001)increased in GH co-treatment group.Conclusions GH is suggested a beneficial impact on oocyte quality and transferrable embryos with PPOS protocol in IVF.Its adjuvant administration can be proposed as an optional therapeutic strategy in women with low prognosis.
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Objective To observe the impacts of acupuncture and moxibustion on kelch like epichlorohydrin related protein 1(Keap1)and nuclear factor E2 related factor 2(Nrf2)signal pathway in kidney tissue of cisplatin(DDP)-induced kidney injury model mice,and to explain the protective mechanism of acupuncture and moxibustion on improving kidney injury caused by DDP.Methods Forty SPF male KM rats were randomly divided into 4 groups,with 10 rats in each group.One day before the start of treatment,the three groups of mice outside the blank group were intraperitoneally injected with cisplatin 10 mg·kg-1 according to their body weight,and the blank group was injected with the same dose of 0.9%NaCl solution.The model was established 24 hours later.Both acupuncture group and moxibustion group selected"Dazhui"(GV14),"Ganshu"(BL18),"Shenshu"(BL23)and"Zusanli(Housanli)"(ST36)for acupuncture and moxibustion respectively,once a day for 5 days.The other two groups were fixed every day without treatment.After fasting for 1 day,the contents of BUN,Scr,CysC and NGAL in serum and Keap1 and Nrf2 in renal tissue were detected by ELISA;Western blot and Real-time PCR were used to detect the protein expression and gene transcription of Keap1 and Nrf2 in the kidney tissue of mice in each group.Results Compared with the blank group,the content of Keap1 protein,protein expression and relative expression of mRNA in the model group increased(P<0.05),the content of Nrf2 protein,protein expression decreased(P<0.05),Nrf2 relative expression of mRNA increased(P<0.05);Compared with the model group,the content of Keap1 protein,the expression of Keap1 protein and the relative expression of Keap1 mRNA in the kidney of mice in the acupuncture and moxibustion groups decreased(P<0.05);Nrf2 protein content,protein expression and relative mRNA expression increased(P<0.05).Conclusion Acupuncture and moxibustion can improve the renal function of DDP renal injury model mice and enhance their antioxidant stress ability,so as to improve the renal injury caused by DDP chemotherapy.Its mechanism may be related to Keap1/Nrf2 signal pathway.
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Objective:To explore the effects of breast cancer mesenchymal stem cells (BC-MSC) on the proliferation and migration of breast cancer MCF-7 cells and the related mechanisms.Methods:The resected cancer tissues and paracancerous tissues were taken from breast cancer patients after surgery, and the bone marrow samples of healthy people were selected. BC-MSC, breast cancer paracancerous mesenchymal stem cells (BCN-MSC) and bone marrow mesenchymal stem cells (BM-MSC) of healthy people were isolated and cultured by tissue adhesion method, and their differentiation ability was induced by the addition of osteogenic and lipogenic induction, and their surface markers were detected by flow cytometry. The supernatants of BC-MSC, BCN-MSC and BM-MSC of healthy people cultured for 48 h were collected and used for the culture of MCF-7 cells as BC-MSC group, BCN-MSC group and BM-MSC group, respectively, and the control group was the conventional cultured MCF-7 cells. The proliferation ability of MCF-7 cells in each group was detected by methyl thiazol tetrazolium (MTT) assay, the clone formation ability of MCF-7 cells was detected by plate cloning assay, the migration ability of MCF-7 cells was detected by Transwell assay, and the mRNA relative expressions of interleukin (IL)-6 and epithelial mesenchymal transition (EMT)-related genes (E-cadherin, vimentin, snail) were detected by quantitative real-time fluorescence polymerase chain reaction (qRT-PCR) in MCF-7 cells. Western blotting was used to detect expressions of p-STAT3, E-cadherin, vimentin and snail proteins in MCF-7 cells. Luminex liquid microarray technology was used to detect cytokine levels in culture supernatants of different mesenchymal stem cells (MSC). IL-6 neutralizing antibody was added into the supernatant of BC-MSC, MCF-7 cells were cultured with the supernatant (BC-MSC+IL-6 neutralizing antibody group), and then the proliferation and migration abilities of MCF-7 cells were tested, as well as the expression changes of related genes and proteins.Results:BC-MSC, BCN-MSC and BM-MSC were successfully isolated; BC-MSC had positive expressions of CD29, CD44 and CD90 and negative expressions of CD14, CD34 and CD45, which were in line with the characteristics of MSC. MTT assay showed that the absorbance values of MCF-7 cells cultured for 48 h in the control group, BC-MSC group, BCN-MSC group and BM-MSC group were 0.31±0.02, 0.54±0.03, 0.43±0.02 and 0.42±0.02, respectively, and the difference was statistically significant ( F = 56.52, P < 0.05); the results of plate cloning experiments showed that the number of clones in each petri dish of the four groups were 180±9, 439±17, 319±16 and 306±19, respectively, and the difference was statistically significant ( F = 222.70, P < 0.05); Transwell assay showed that the numbers of membrane-penetrating cells in the four groups were 6.5±1.0, 23.2±2.4, 16.0±1.3 and 14.8±2.0, respectively, with the statistically significant difference ( F = 49.44, P < 0.05); qRT-PCR assay showed that the relative expressions of IL-6 mRNA in the control group, BC-MSC group, BCN-MSC group and BM-MSC group were 1.07±0.11, 13.79±3.80, 6.68±1.66 and 6.12±1.52, respectively, and the difference was statistically significant ( F = 107.60, P < 0.05), and the relative expression of E-cadherin mRNA in MCF-7 cells of BC-MSC, BCN-MSC and BM-MSC groups was lower than that of the control group, while the relative expressions of vimentin and snail mRNA were higher than those of the control group, and the differences were statistically significant (all P < 0.05). Western blotting assay showed that the relative expression of E-cadherin mRNA in MCF-7 cells of BC-MSC, BCN-MSC and BM-MSC groups was lower than that of the control group. Western blotting showed that the level of E-cadherin protein in BC-MSC, BCN-MSC and BM-MSC groups was lower than that in the control group, and the levels of vimentin and snail proteins were higher than those in the control group; Luminex liquid microarray technology showed that the content of IL-6 cytokine in the supernatants of BC-MSC, BCN-MSC and BM -MSC cultures were higher, and the relative expressions were 1.75±0.21, 1.00±0.10 and 0.96±0.08, respectively, and the difference was statistically significant ( F = 43.22, P < 0.05). The results of MTT assay showed that the absorbance values of MCF-7 cells in BC-MSC group and BC-MSC+IL-6 neutralizing antibody group were 0.56±0.05 and 0.42±0.04, respectively, and the difference was statistically significant ( t = -3.11, P < 0.05); the results of Transwell assay showed that the numbers of membrane-penetrating cells in the two groups were 30.3±1.5 and 17.3±2.1, respectively, and the difference was statistically significant ( t = -7.12, P < 0.05); qRT-PCR assay showed that the relative expressions of E-cadherin mRNA were 0.44±0.05 and 0.76±0.05 ( t = 6.40, P < 0.01), the relative expressions of vimentin mRNA were 2.90±0.21 and 1.79±0.21 ( t = 5.29, P < 0.01), and the relative expressions of snail mRNA were 3.20±0.20 and 1.91±0.30 ( t = 2.16, P < 0.01); Western blotting assay showed that the degrees to down-regulate the expression of E-cadherin protein and up-regulate the expressions of vimentin and snail proteins in the BC-MSC+IL-6 neutralizing antibody group were weakened compared with the BC-MSC group. Conclusions:BC-MSC can promote the proliferation and migration of breast cancer MCF-7 cells probably through activating IL-6-STAT3 signaling pathway-induced EMT by its secretion of IL-6.
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Objective:To evaluate the effect of nimodipine on postoperative cognitive function in elderly patients undergoing carotid endarterectomy.Methods:Eighty-two American Society of Anesthesiologists physical status Ⅱ or Ⅲ patients of both sexes, aged 65-80 yr, scheduled for elective carotid endarterectomy under general anesthesia, were divided into 2 groups ( n=41 each) using a random number table method: control group (group C) and nimodipine group (group N). Nimodipine 7.5 μg·kg -1·h -1 was intravenously infused starting from the beginning of surgery until the end of surgery in group N, while the equal volume of normal saline was given in group C. Before infusing nimodipine (T 1), before placing the the shunt (T 2), at 10 min after placing the the shunt (T 3) and at 10 min after releasing carotid artery (T 4), blood samples were taken from the radial artery and jugular bulb for blood gas analysis.Jugular venous blood oxygen content, arterio-jugular difference of oxygen content, and cerebral oxygen extraction ratio were calculated.The concentrations of S100β protein in serum of the jugular bulb were measured by enzyme-linked immunosorbent assay.Cognitive function was assessed using the Montreal Cognitive Assessment (MoCA) Scale (Chinese version) at 1 day before surgery and 1, 3 and 7 days after surgery, and the occurrence of cognitive dysfunction (MoCA score<26) was recorded within 7 days after operation. Results:Compared with group C, MoCA scores were significantly increased at each time point after surgery, and the incidence of cognitive dysfunction was decreased (27% vs.17%), and the jugular venous blood oxygen content was increased, and arterio-jugular difference of oxygen content, cerebral oxygen extraction ratio, and concentrations of serum S100β protein were decreased at T 2-4 in group N ( P<0.05). Conclusions:Nimodipine can improve the cognitive function after carotid endarterectomy, which may be related to the improvement in intraoperative cerebral oxygen metabolism and reduction of brain injury in elderly patients.
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Objective:To evaluate the effect of low-dose esketamine on postoperative delirium (POD) in elderly frail patients undergoing laparoscopic radical resection of the gastrointestinal tumor.Methods:Ninety-four American Society of Anesthesiologists physical status Ⅱ or Ⅲ frail patients of both sexes, aged 65-80 yr, with body mass index of 18.5-30.0 kg/m 2 and preoperative Fried frailty phenotype scale score≥3, scheduled for elective laparoscopic radical resection of the gastrointestinal tumor under general anesthesia, were divided into 2 groups ( n=47 each) using a random number table method: control group (group C) and low-dose esketamine group (group K). In group K, esketamine 0.5 mg/kg was given during induction of anesthesia, and esketamine 0.25 mg·kg -1·h -1 was continuously infused during operation until the end of operation.In group C, the equal volume of normal saline was given at the corresponding time point.After induction of anesthesia and before skin incision (T 1), at 1 day after operation (T 2) and at 3 days after operation (T 3), blood samples from the internal jugular vein were collected for determination of the concentrations of S100β protein and neuron-specific enolase (NSE) in serum by enzyme-linked immunosorbent assay.The intraoperative consumption of propofol, remifentanil and sufentanil and use of vasoactive drugs were recorded.POD was evaluated by Confusion Assessment Method within 3 days after operation. Results:Compared with the baseline at T 1, the concentrations of serum S100β and NSE were significantly increased at T 2, 3 in both groups ( P<0.05). Compared with group C, the incidence of POD was significantly decreased (30% vs.13%), the intraoperative consumption of propofol and remifentanil was decreased, and concentrations of serum S100β protein and NSE were decreased at T 2, 3 in group K ( P<0.05). Conclusions:Low-dose esketamine can decrease the occurrence of POD in elderly frail patients undergoing laparoscopic radical resection of gastrointestinal tumor.
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Objective:To evaluate the improved efficacy of ultrasonography-guided superior laryngeal nerve block (SLNB) in elderly patients undergoing short surgery with general anesthesia.Methods:Sixty-four patients of both sexes, aged≥65 yr, of American Society of Anesthesiologists physical status Ⅱ or Ⅲ, scheduled for elective percutaneous balloon compression under general anesthesia, were divided into 2 groups ( n=32 each) using a random number table method: control group (group C) and ultrasound-guided SLNB group (group S). The patients received bilateral SLNB under ultrasound guidance, and 1% lidocaine 3 ml was injected on each side in group S, while the equal volume of normal saline was given instead in group C. Anesthesia was induced with midazolam, sufentanil, etomidate and mivacurium, and then the patients were mechanically ventilated after endotracheal intubation.Anesthesia was maintained with propofol, remifentanil, and sevoflurane.Cardiovascular response to endotracheal intubation was defined as SBP or HR increased by more than 30% of baseline from the time point immediately after intubation to 2 min after intubation, and the occurrence was recorded.Venous blood samples were collected to detect the plasma concentrations of norepinephrine and cortisol before anesthesia induction and at 5 min after intubation.The development of bucking was recorded during emergence, and the time of tracheal extubation and occurrence of sore throat, throat numbness and hoarseness after tracheal extubation were recorded. Results:Compared with group C, the incidence of cardiovascular response to endotracheal intubation was significantly decreased, the plasma concentrations of norepinephrine and cortisol were decreased at 5 min after intubation, and the incidence of bucking during emergence and sore throat after tracheal extubation was decreased in group S ( P<0.05). Conclusions:Ultrasound-guided SLNB can inhibit the stress response during endotracheal intubation and reduce the occurrence of adverse events during emergence in elderly patients undergoing short surgery with general anesthesia.
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@#Objective To explore the independent risk factors for postoperative retention of urinary catheters in the ward of lung tumor patients due to urinary retention under the concept of enhanced recovery after surgery (ERAS). Methods Seventy-five patients with lung tumors who had urinary catheters left in the postoperative ward between June 2019 and August 2019 were selected as a case group, and 75 patients with lung tumors who did not have urinary catheters in the perioperative period as a control group. Independent risk factors for indwelling urinary catheters in the postoperative ward were screened by univariate and multiple-variate logistic stepwise regression analysis. Results There were 45 males and 30 females in the case group with an average age of 55.33±10.78 years, 28 males and 47 females in the control group with an average age of 57.12±10.06 years. Univariate analysis showed that gender, operative time>2 h, intraoperative fluid volume≥1 200 mL, and fluid volume within 6 h of returning to the ward after surgery>1 200 mL were associated with the occurrence of indwelling urinary catheters in patients with lung tumors in postoperative wards (P<0.05). Multiple-variate logistic regression showed that male (OR=2.311, 95%CI 1.173-4.552, P=0.015), infusion volume within 6 h of returning to the ward after surgery>1 200 mL (OR=2.491, 95%CI 1.149-5.401, P=0.021) and intraoperative infusion volume≥1 200 mL (OR=2.105, 95%CI 1.022-4.340, P=0.044) were independent risk factors for postoperative retention of urinary catheters in patients with lung tumors. Conclusion The occurrence of indwelling urinary catheter in lung tumor patients under the ERAS concept is the result of a combination of factors, and patients who are male, have infusion volume>1 200 mL within 6 h of returning to the ward after surgery, and have intraoperative infusion volume≥1 200 mL are the high-risk group for postoperative ward indwelling urinary catheter, and health care personnel should strengthen the assessment and observation, provide targeted health education, appropriately control the perioperative fluid volume, and take other measures to reduce the occurrence of indwelling urinary catheters due to urinary retention postoperatively in ward.
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AIM: To investigate the sensitization of flavonoids from Tetrastigma hemsleyanum (FTH) on gefitinib (GEF)-resistant lung adenocarcinoma cells. METHODS: The viabilities of A549 and A549/GR cells treated with FTH and GEF were detected by MTT method. The apoptotic rates and cell cycles of A549/GR cells treated with FTH and GEF were detected by Flow cytometry. The anti-tumor effects of flavonoids from FTH and GEF were assayed in A549/GR tumor-bearing mice. The expressions of proteins (PTEN, PI3K, p-PI3K, AKT, p-AKT) were detected by Western blot analysis. RESULTS: Compared with GEF group, FTH significantly enhanced the inhibition of GEF on the proliferation of A549/GR cells (P< 0.05). Combination with FTH and GEF significantly increased the apoptosis of A549/GR cells which were arrested at the G
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Objective:To determine the dose-effect relationship of nalbuphine preventing injection pain of medium plus long chain triglyceride propofol in pediatric patients undergoing gastroenteroscopy.Methods:Pediatric patients, aged 3-8 yr, of American Society of Anesthesiologists physical statusⅠ or Ⅱ, scheduled for elective gastroenteroscopy, were enrolled in the study.The doses of nalbuphine were determined by up-down sequential allocation, nalbuphine 0.2 mg/kg was injected intravenously in the first child, and 5 min later medium plus long chain triglyceride propofol 2.5 mg/kg was given intravenously.Ambesh 4-point method was used to evaluate the injection pain of propofol.When the prevention of injection pain was ineffective, the dose of nalbuphine was increased in the next patient, otherwise the dose was reduced, and the difference between the two successive doses was 0.01 mg/kg.This process was repeated until the 7th turning point occurred.The ED 50 and ED 95 of nalbuphine and 95% confidence interval (CI) preventing injection pain of propofol were calculated by Probit regression. Results:The ED 50 and ED 95 (95% CI) of nalbuphine preventing medium plus long chain triglyceride propofol injection pain were 1.57 (1.50-1.62) and 1.71 (1.64-2.05) mg/kg, respectively. Conclusion:The ED 50 and ED 95 of nalbuphine preventing injection pain of medium plus long chain triglyceride propofol are 1.57 and 1.71 mg/kg, respectively, in pediatric patients undergoing gastroenteroscopy.
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OBJECTIVE@#To investigate the effects of etomidate on electrophysiological properties and nicotinic acetylcholine receptors (nAChRs) of ventral horn neurons in the spinal cord.@*METHODS@#The spinal cord containing lumbosacral enlargement was isolated from 19 neonatal SD rats aged 7-12 days. The spinal cord were sliced and digested with papain (0.18 g/30 mL artificial cerebrospinal fluid) and incubated for 40 min. At the ventral horn, acute mechanical separation of neurons was performed with fire-polished Pasteur pipettes, and perforated patch-clamp recordings combined with pharmacological methods were employed on the adherent healthy neurons. In current-clamp mode, the spontaneous action potential (AP) of the ventral horn neurons in the spinal cord was recorded. The effects of pretreatment with different concentrations of etomidate on AP recorded in the ventral horn neurons were examined. In the voltage-clamp mode, nicotine was applied to induce inward currents in the ventral horn neurons, and the effect of pretreatment with etomidate on the inward currents induced by nicotine were examined with different etomidate concentrations, different holding potentials and different use time.@*RESULTS@#The isolated ventral horn neurons were in good condition with large diverse somata and intact processes. The isolated spinal ventral horn neurons (=21) had spontaneous action potentials, and were continuously perfused for 2 min with 0.3, 3.0 and 30.0 μmol/L etomidate. Compared with those before administration, the AP amplitude, spike potential amplitude and overshoot were concentration-dependently suppressed ( < 0.01), and spontaneous discharge frequency was obviously reduced ( < 0.01, =12). The APs of the other 9 neurons were completely abolished by etomidate at 3.0 or 30 μmol/L. At the same holding potential (VH=-70 mV), pretreatment with 0.3, 3.0 or 30.0 μmol/L etomidate for 2 min concentration-dependently suppressed the current amplitude induced by 0.4 mmol/L nicotine ( < 0.01, =7). At the holding potentials of - 30, - 50, and - 70 mV, pretreatment with 30.0 μmol/L etomidate for 2 min voltage-dependently suppressed the current amplitude induced by 0.4 mmol/L nicotine ( < 0.01, =6 for each holding potential). During the 6 min of 30.0 μmol/L etomidate pretreatment, the clamped cells were exposed to 0.4 mmol/L nicotine for 4 times at 0, 2, 4, and 6 min (each exposure time was 2 s), and the nicotinic current amplitude decreased gradually as the number of exposures increased. But at the same concentration, two nicotine exposures (one at the beginning and the other at the end of the 6 min pretreatment) resulted in a significantly lower inhibition rate compared with 4 nicotine exposures ( < 0.01, =6).@*CONCLUSIONS@#etomidate reduces the excitability of the spinal ventral neurons in a concentration-dependent manner and suppresses the function of nAChR in a concentration-, voltage-, and use-dependent manner.
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Animales , Ratas , Animales Recién Nacidos , Etomidato , Neuronas , Técnicas de Placa-Clamp , Médula EspinalRESUMEN
Objective To analyze the anatomical characteristics of the upper airway in Pierre Robin sequence pediatric patients with difficult laryngoscopy using the computed tomography-based three-dimensional reconstruction.Methods Fifty pediatric patients of both sexes with Pierre Robin sequence,aged 10-101 days,weighing 2.0-6.3 kg,of American Society of Anesthesiologists physical status Ⅲ,scheduled for elective mandibular distraction osteogenesis under general anesthesia,were enrolled in this study.Cone beam CT scan was performed to obtain upper airway anatomy information during the natural sleep before operation.Images were imported into medical engineering software MIMICS 17.0 to reconstruct the three-dimensional images of the oral and maxillofacial bones and airways.The related anatomical parameters were measured,including the distance between the alveolar ridge of the upper central incisor and root of the epiglottis (D1),distance between the root of the epiglottis and midpoint of glottis (D2),distance between the bilateral lower edge of the mandible and midpoint of glottis (D3),distance between the alveolar ridge of the lower central incisor and the lower edge of the mandible (D4),length of the mandibular ramus (D5),length of the mandible body (D6),and length of the total mandible (D7),angle between lines D1 and D2 (angle 1),the angle between line D2 and the alveolar ridge of the upper central incisor to the midpoint of glottis (angle 2),the angle between lines D3 and D4 (angle 3),the angle of the point of the upper central incisor alveolar ridge to the trailing edge of the hard palate and then to the root of epiglottis (angle 4),the angle of bilateral mandible (angle 5),the angle of the point of gnathion to the two gonions (angle 6),the airway cross-sectional area at the tip of epiglottis,volume of oral cavity,volume of velopharyngeal cavity,and volume of glossopharyngeal cavity.Fiberoptic bronchoscope-guided endotracheal intubation was performed under topical anesthesia with lidocaine.Propofol,sufentanil and cis-atracurium were intravenously injected to induce anesthesia after successful intubation,and then the pediatric patients were sent to the operating room.Anesthesia was maintained by inhalation of sevoflurane.The exposure of glottis was observed with a laryngoscope.Pediatric patients were divided into difficult laryngoscopy group (group A) and non-difficult laryngoscopy group (group B) according to whether they presented with difficult laryngoscopy (Cormack-Lehane classification Ⅲ or Ⅳ).Results Compared with group B,the airway cross-sectional area at the tip of epiglottis and in the volume of velopharyngeal cavity were decreased (P＜0.05),and no significant change was found in D1,D2,D3,D4,D5,D6,D7,angle 1,angle 2,angle 3,angle 4,angle 5,angle 6,volume of oral cavity or volume of glossopharyngeal cavity in group A (P＞0.05).Conclusion The three-dimensional CT images of the upper airway show characteristic changes in Pierre Robin sequence pediatric patients with difficult laryngoscopy,and the main manifestations are the decrease in the airway section area and in the volume of the palatopharyngeal cavity at the tip of the epiglottis.
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Objective: To investigate the repeatability of different ROI seletion methods on histogram analysis parameters of enhanced T1-weighted images of high-grade gliomas (HGGs). Methods: Four ROI selecting methods (small ROI method, contour ROI method, threshold ROI method and volume ROI method) were used to measure the histogram parameters on enhanced T1WI of 45 HGGs patients. The consistency between 2 observers and the differences of histogram parameters were evaluated. Results: The consistent of the mean, standard deviation and kurtosis values measured using volume, contour and threshold ROI methods between 2 observers were good (ICC≥0.80), and of the skewness values were moderate to good (ICC=0.73-0.90). The consistent of histogram characteristic parameters measured using small ROI method between 2 observers were low to medium (ICC=0.30-0.69). Histogram characteristic parameters measured with volume ROI method and contour ROI method had no significant difference (all P>0.05), but the average value, skewness value and kurtosis value measured with threshold ROI method were higher than those with volume and contour ROI method (all P<0.05), and the standard deviation of threshold ROI method was lower than that of volume ROI method and contour ROI method (both P<0.05). Standard deviation measured with small ROI method was lower than that of volume ROI method and contour ROI method (all P<0.05). Conclusion: The consistency of histogram characteristic parameters of HGGs on enhanced T1WI measured with volume ROI method is the highest, and different ROI selection methods have some impact on the measurement results of histogram characteristic parameters of enhanced T1WI.
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Objective@#To establish a real-time PCR (RT-PCR) assay for detecting mRNA expression of killer cell immunoglobulin-like receptor (KIR) 2DS1 gene( KIR2DS1 ) on the surface of natural killer (NK) cells, and evaluate its performance. @*Methods@#A total of 57 recipient-donor pairs of allogeneic hematopoietic stem cell transplantation (Allo-HSCT) were enrolled in this study. The specific primers and probe of KIR2DS1 gene were designed for Taqman-MGB fluorescence quantitative PCR detection system. The performance parameters of the detecting system, such as coincidence rate, repeatability, sensitivity, scope of application of the instrument and reproducibility of operation technicians were evaluated and validated. @*Results@#The KIR-SSO Genotyping Test was used as the gold standard. The results of 35 samples showed the accuracies of self-built method were all 100% for both of positive and negative KIR2DS1 . Three samples with high, median and low value of Ct values were used to verify the repeatability. The coefficients of variation of intra-assay and inter-assay were ranged from 0.09% to 0.46% and 0.71% to 1.13% respectively. The sensitivity of the established method was up to 10 2 copies/μL at least. The coefficients of variation of the three samples with sensitivity of 10 2 copies/μL were 5.37%, 2.71% and 5.51% in five repeated tests respectively. The regression analysis for the samples measured by ABI-7500 and LC-480 fluorescence quantitative PCR instrument showed regression equation was Y=0.973 6X+0.118 3 (R 2 =0.961 9, R 2 ＞0.95). The reproducibility of 10 samples with positive KIR2DS1 operated by two technicians showed that the biases were all less than ±5%. @*Conclusion@#A TaqMan-MGB real-time PCR assay for detection of mRNA expression of KIR2DS1 gene was established successfully with fine performance.
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<p><b>OBJECTIVE</b>To explore the role of β2-nicotinic acetylcholine receptor (β2-nAChR) in the development of γ- aminobutyric acid A type receptors (GABA-Rs) in hippocampal CA1 and CA3 pyramidal neurons of mice.</p><p><b>METHODS</b>The hippocampal CA1 and CA3 pyramidal neurons were acutely isolated from β2-nAChR gene knockout (β2-KO group) mice. GABA currents in CA1 and CA3 pyramidal neurons were induced with the selective GABA-R agonist muscimol and recorded using perforated patch-clamp recording technique. The GABA currents of CA1 and CA3 pyramidal neurons were tested for their equilibrium potentials (Es) and kinetic parameters and were compared with the measurements in wild-type mice (WT group).</p><p><b>RESULTS</b>The mean E of CA1 neurons (=7) of β2-KO mice (=4) was -31.7±3.5 mV, showing an obvious depolarizing shift compared with the WT mice ( < 0.05); the mean E of CA3 neurons (=4) was -16.1±4.6 mV, also showing a depolarizing shift ( < 0.01). The difference in the Es between CA3 and CA1 neurons in β2-KO mice, but not in WT mice, was significant ( < 0.05). The GABA-R desensitization was significantly slowed down in both CA1 and CA3 neurons of β2-KO mice, with decay time of 2.2±0.2 s and 3.2±0.1 s, respectively, significantly longer than those in WT mice (1.6±0.1 s and 2.3±0.1 s, respectively; < 0.05).</p><p><b>CONCLUSIONS</b>β2-containing nAChRs may promote the functional maturation of GABA-R in CA1 and CA3 pyramidal cells in mouse hippocampus.</p>
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Objective To explore the effects of heating intravenous fluid infusion and blood transfusion based on guidelines in severe trauma patients with hypothermia. Methods A total of 40 severe trauma patients with hypothermia admitted from July 2014 to December 2015 were enrolled as the control group treated with routine measures to maintain the body temperature at normothermia by such as electrical heating blanket; other 40 severe casualties with hypothermia admitted from January 2016 to July 2017 were recruited as the warming up group treated with heating intravenous fluid infusion and blood transfusion by hot water bath in addition to the routine measures for keeping body temperature at normothermia. The differences in core body temperature, prothrombin time, activated partial thromboplastin time, incidence of shivering and mortality rate were compared between the two groups. Results There was statistically signifi cant difference in core body temperature at 0.5 h, 1.0 h, 1.5 h, 3.0 h between the two groups (P<0.05). Though the prothrombin time and shivering were improved after warming up in both groups, and there were significant differences in prothrombin time at 3.0 h after warming up and the incidence of shivering between two groups(P<0.05).There was no signifi cant difference in mean arterial pressure at all seven intervals between two groups. Conclusion The heating intravenous fl uid infusion and blood transfusion had remarkable effects to prevent hypothermia, improves blood coagulation and reduced the incidence of shivering to provide more simple and convenient warming up intervention for clinical practice.
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Crude glycerol is the main by-product of biodiesel production. A few microorganisms can transfer crude glycerol to 1,3-propanediol (1,3-PD) that is an important chemical material. There exist many limitations such as substrate inhibition, product inhibition when wild strains are used in 1,3-PD biosynthesis. In this review, based on the microbial transformation of 1,3-propanediol from glycerol and its limitations, some strategies using genetic engineering such as knockout or gene overexpression were summarized. The latest research progresses in biosynthesis of 1,3-propanediol from glycerol by genetically engineered strains are discussed.
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Objective@#To analyze the distribution and proportion of donor-specific activated killer cell immunoglobulin like receptor (aKIR) genes and their clinical application values in unrelated allogeneic hematopoietic stem cell transplantation (allo-HSCT) .@*Methods@#Retrospective analyses of KIR genotyping using polymerase chain reaction with sequence specific primers (PCR-SSP) were performed in 216 pairs of donors and recipients.@*Results@#The frequency of donor-specific KIR genes was 53.7% (116/216) in 216 patients receiving unrelated allo-HSCT, with the frequency of 78.3% (112/143) in the KIR genes mismatched group and 5.5% (4/73) in matched group. Of the 116 patients with detectable donor-specific KIR genes, 99.1% (115/116) patients had various donor-specific aKIR genes. Among 55 pairs of donors’ KIR-Bx genotype and patients’ KIR-AA genotype group, the most commonly observed genotypes were Bx1, Bx2, Bx3, Bx4, in which the donor-specific KIR genes were respectively KIR 3DS1, 2DL5A, 2DS5, 2DS1; KIR 3DS1, 2DL5A, 2DS3, 2DS1; KIR 2DS2, 2DL2; KIR 2DS2, 2DL2, 3DS1, 2DL5A, 2DS5, 2DS1. Of 44 pairs of donors’ KIR-AA genotype and patients’ KIR-Bx (AB) genotype group, 36.4% (16/44) recipients had donor-specific KIR2DS4 (FUL) gene. In 143 pairs of KIR mismatched group, the frequencies of donor-specific KIR genes were KIR2DS1 (35.7%) , KIR3DS1 (32.9%) , KIR2DS5 (29.4%) , KIR2DS4 (FUL) (25.9%) , KIR2DL2 (25.2%) , KIR2DS2 (24.5%) , KIR2DS3 (21.7%) and KIR3DL1 (8.4%) , respectively.@*Conclusion@#The donor-specific aKIR genes mainly existed in KIR mismatched group after unrelated allo-HSCT, and the different pairs of donors’ and patients’ KIR genotypes led to the diverse donor-specific aKIR. But there were higher specific aKIR genes in higher frequency of KIR AA, Bx1, Bx2, Bx3, Bx4 genotypes. All these can provide the experimental basis for studying the role of the donor-specific aKIR genes on the prognosis of HSCT.
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Objective@#To investigate the effect of curcumin on the apoptosis and autophagy of human gastric cancer cells with different degree of differentiation.@*Methods@#Gastric cancer cell lines BGC-823 and MKN-28 were treated with curcumin at different concentrations. The effect of curcumin on cell proliferation was measured by MTT assay. Apoptosis was assessed by flow cytometry. Autophagy status was analyzed by acridine orange staining. The expression levels of apoptotic and autophagy-related proteins were detected by Western blot.@*Results@#The cell viability of BGC-823 and MKN-28 was inhibited by curcumin in a time- and dose-dependent manner. At 48 h after treatment, the IC50 value of BGC-823 (15.18 μmol/L) was close to that of MKN-28 (15.84 μmol/L), and the difference was not statistically significant (P=0.513). Meanwhile, flow cytometry showed that curcumin induced the apoptosis of gastric cancer cells in a dose-dependent manner. Western blot results showed that the expression of pro-apoptotic proteins bax, active-caspase-3 and active-caspase-9 was significantly increased in BGC-823 and MKN-28 cells, whereas that of the anti-apoptotic protein bcl-2 was strikingly reduced. In addition, the formation of acidic vesicular organelles in cytoplasm, conversion of LC3-Ⅰ to LC3-Ⅱ and increased levels of autophagy-related proteins Beclin1, Atg7 and Atg5-Atg12 were observed in curcumin-treated cells. Moreover, activation of PI3K/Akt/mTOR signaling pathway was also significantly suppressed after curcumin treatment. Blocking autophagy by adding the autophagy inhibitor 3-methyladenine (3-MA) significantly promoted the apoptotic cell death induced by curcumin.@*Conclusions@#Curcumin induces apoptosis and protective autophagy in human gastric cancer cells in vitro. Curcumin combined with autophagy inhibitor may provide a more effective strategy for its clinical application.
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Objective@#To study the clinical effect of high pressure oxygen and Butylphthalide in the recovery of cerebral metabolism after carbon monoxide poisoning.@*Methods@#84 patients treated from May 2014 to May 2016 in our hospital were selected. The subjects were randomly and equally divided into two groups. The control group adopted the conventional therapy and high pressure oxygen; on the basis, the observation group also took Butylphthalide. The clinical effect, duration of coma, recovery of consciousness, incidence rate of delayed encephalopathy was observed. After 1m of treatment, the HDS point was evaluated.@*Results@#The total effective rate of control group (76.19%, 32/42) was lower than that of observation group (95.24%, 40/42) (P<0.05) . The duration of coma for observation group was shorter than that of control group. The percentage for patients with recovery of consciousness and incidence rate of delayed encephalopathy for observation group was better than that of control group (P<0.05) . The HDS point for observation group was even higher than that of control group (P<0.05) .@*Conclusion@#The high pressure oxygen and butylphthalide can improve the clinical effective rate, shorten the duration of coma and promote the patient’s recovery of consciousness. It is worthy of clinical promotion.
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Objective@#To investigate the effect of mesenchymal stem cells (MSCs) on apoptosis of breast cancer cell line MCF-7 induced by cisplatin (DDP), MSCs derived from breast cancer (BC-MSCs) or adjacent non-cancerous tissues (BN-MSCs) were isolated, cultured and identified.@*Methods@#BC-MSCs and BN-MSCs were isolated and cultured by tissue adherent method. The differentiation potential of BC-MSCs was detected by osteogenic and adipogenic induction, and cell surface markers of BC-MSCs and BN-MSCs were evaluated by flow cytometry. MCF-7 cells were co-treated with DDP and conditioned medium (CM) collected from BC-MSCs and BN-MSCs after being cultured for 48 hours, respectively. Inhibition rate of cell proliferation was evaluated by MTT. Cell apoptosis and viability were detected by MUSE cell analyzer. Cytokines in MSC-CM were detected by Luminex liquid chip. Interleukin 6 (IL-6) mRNA expressions in MCF-7 cells with different treatment were detected by RT-PCR.@*Results@#The morphology of BC-MSCs and BN-MSCs successfully isolated and cultured was uniform fibroblast-like clusters under the microscope. These cells expressed high levels of CD29 and CD44, but neither CD14 nor CD34 were detected. MSCs could also differentiate into osteoblasts and adipocytes after specific induction. After treatment with 2.5, 5, 10, 20, 40 and 80 μmol/L DDP, the inhibitory rates of proliferation of MCF-7 cells in DDP group were (17.33±2.00)%, (22.37±0.73)%, (30.77±1.23)%, (44.93±1.27)%, (62.03 ±1.97)% and (73.93±1.10)%, respectively. While the inhibitory rates of DDP+ BC-MSCs group were (8.27±0.63)%, (11.50±1.30)%, (20.57±0.93)%, (32.60 ±1.90)%, (52.27±0.73)% and (62.13±2.17)%, respectively. The inhibitory rates of DDP+ BN-MSCs group were (12.90±1.60)%, (16.53±2.87)%, (25.90±1.50)%, (39.40±2.40)%, (57.40±0.70)% and (69.03±1.07)%, respectively. The inhibitory rates of DDP+ BC-MSCs group were significantly lower than those of DDP group (P<0.05). The apoptotic rates of MCF-7 cells in DDP group, DDP+ BC-MSCs group and DDP+ BN-MSCs group were (47.77±1.98)%, (29.20±2.12)% and (37.92±2.21)%, respectively. The apoptotic rates of DDP group was significantly higher than that of DDP+ BC-MSCs group (P<0.05). The cell viabilities of MCF-7 in DDP group, DDP+ BC-MSCs group and DDP+ BN-MSCs group were 0.52±0.02, 0.72±0.02 and 0.64±0.02, respectively. The cell viability of DDP group was significantly lower than that of DDP+ BC-MSCs group (P<0.05). The result of Luminex liquid chip analysis showed that, the level of IL-6 in BC-MSCs group increased 2.50±0.68 fold when compared with BN-MSCs group (P<0.05). The relative expressions of IL-6 mRNA in DDP group and DDP+ BC-MSCs group were 1.02±0.10 and 7.58±0.55, respectively, with a statistically significant difference (P<0.01). The apoptotic rates of MCF-7 cells in DDP+ BC-MSCs group with or without IL-6 neutralizing antibody were (27.41±1.95)% and (42.45±2.87)%, respectively, with a statistically significant difference (P<0.05). The cell viabilities of MCF-7 cells in DDP+ BC-MSCs group with or without IL-6 neutralizing antibody were (72.40±2.60)% and (59.76±3.89)%, respectively, with a statistically significant difference (P<0.05).@*Conclusions@#BC-MSCs and BN-MSCs have been isolated and cultured successfully. Compared with BN-MSCs, BC-MSCs could attenuate the effect of DDP on MCF-7 cells, evidently decrease the apoptosis and increase the proliferation and vitality in an IL-6 dependent manner.