Objective:To discuss the effect of prostaglandin E2(PGE2),a pyrogenic mediator,on the discharge activity of warm-sensitive neurons(WSNs)in median preoptic nucleus(MnPO)of hypothalamus in the female mice,and to clarify its mechanism.Methods:Coronal brain slices of MnPO were prepared from the female mice.The slices were then perfused with artificial cerebrospinal fluid(ACSF)containing synaptic transmission blockers(STBs).The discharge frequency was monitored using the patch clamp technique while changing the temperature of the perfusate to identify the WSNs.A total of 32 MnPO WSNs were divided into base line group(n=32)and PEG2(n=32).The patch clamp technique was employed to monitor the discharge frequencies of MnPO WSNs following the perfusion of ACSF and 1 μmol·L-1 PGE2,respectively.The MnPO WSNs with good activity and significant change in discharge frequency after PGE2 perfusion were divided into PGE2 receptor E-series prostaglandin receptrol(EP)1 antagonist(EP1 ant)+PGE2 group(n=7),EP3 ant+PGE2 group(n=7),and EP4 ant+PGE2 group(n=7).The patch clamp technique was used to monitor the discharge frequencies of MnPO WSNs following the perfusion of 3 μmol·L-1 EP1 ant and 1 μmol·L-1 PGE2 mixture,10 μmol·L-1 EP3 ant and 1 μmol·L-1 PGE2mixture,and 10 μmol·L-1 EP4 ant and 1 μmol·L-1 PGE2 mixture,respectively.Resluts:After perfuson of the ACSF containing STBs,a total of 188 MnPO neurons from the female mice with an intrinsic temperature sensitivity coefficient(m value)were identified;out of these,32 neurons had an m value of ≥0.8 and were identified as MnPO WSNs,accounting for approximately 17％of all recorded neurons.Compared with baseline discharge frequency,the discharge frequency of MnPO WSNs after addition of PGE2 was decreased(P＜0.05).Compared with PGE2 group,the percentage change in discharge frequency of MnPO WSNs in EP3 ant+PGE2 group was significantly decreased(P＜0.05).Compared with PGE2 group,the percentage change in discharge frequency of MnPO WSNs in EP1 ant+PGE2 group had no significant difference(P＞0.05).Compared with PGE2 group,the percentage change in discharge frequency of MnPO WSNs in EP4 ant+PGE2 group had no significant difference(P＞0.05).Conclusion:In the female mice,WSNs make up approximately 17％of the total neurons in MnPO.PGE2 can directly inhibit the discharge activity of MnPO WSNs in the female mice through postsynaptic mechanism involving EP3 receptors.

The Choledochal cyst is an extremely rare congenital anomaly of the bile duct. Early cyst resection and Roux-en-Y hepatojejunostomy are the primary surgical methods for treating choledochal cyst. With the emergence of enhanced recovery after surgery, laparoscopic surgery has effectively reduced the incidence of biliary complications and wound infections, but it still does not meet people's requirements for minimally invasive surgery. Robotic surgery system has the potential to enhance surgical precision and the maneuverability of surgeons due to clear surgical visualization and flexible mechanical arms. The authors review the relevant literatures and conduct a Meta-analysis to evaluate the efficacy of robot-assisted surgery and laparoscopic surgery for choledochal cyst.

Humans ; East Asian People ; Psoriasis/drug therapy* ; Antibodies, Monoclonal, Humanized/therapeutic use* ; Treatment Outcome ; Severity of Illness Index

Objective : To investigate the effect of adeno⁃associated virus ( AAV) delivery of short hairpin RNA ( shRNA) against the Ptger3 gene in the lateral parabrachial nucleus (LPB) on the fever induced by microinjection of prostaglandin E2 (PGE2 ) into the LPB and the intraperitoneal injection of lipopolysaccharide (LPS) . Methods: AAV2⁃shRNA⁃Ptger3(EP3) ⅣEGFP ( shRNA⁃EP3) and AAV2⁃ CMV⁃ EGFP ( shRNA⁃control) viruses were constructed and transfected the rat LPB by stereotaxic injection. Four weeks later, the transfection efficiency of AAV viruses was observed by fluorescence microscopy , and the knockdown efficiency was determined by real⁃time PCR of EP3 receptor mRNA on the LPB. The effects of microinjection of saline or PGE2 in the LPB or intraperitoneal injection of LPS on body temperature (Tcore ) and energy expenditure (EE) of shRNA⁃control group and shRNA⁃EP3 group were monitored using an animal monitoring system with temperature telemetry. Results : AAV virus transfecnificant difference in basal body temperature between shRNA⁃control group and shRNA⁃EP3 group. Tcore and EE were briefly and slightly increased after microinjection of saline in the LPB , but there was no significant difference between the two groups. Compared with the shRNA⁃control group , the febrile response induced by LPB PGE2 was attenuated in the shRNA⁃EP3 group (P < 0. 05) . Furthermore , the knockdown of EP3 receptor of LPB also attenuated the LPS⁃induced fever, and the Tcore 5. 5 h post⁃LPS in the shRNA⁃EP3 rats increased compared with the baseline (P < 0. 05) , which was lower than that in the shRNA⁃control rats ( P < 0. 01) . Conclusion EP3 receptor knockdown in LPB attenuates the febrile response induced by microinjection of PGE2 in the LPB and intraperitoneal injection of LPS , suggesting that EP3 receptors of LPB mediate the pyrogenic action of LPB PGE2 and partly participate in LPS⁃induced fever.

Objective : To investigate the effect of Arginine Vasopressin (AVP) on the median preoptic glutamatergic (MnPOVglut2 ) neurons and its mechanism． Methods : Brain slices were prepared from male Vglut2-tdTomato mice．MnPOVglut2 neurons expressing red fluorescent protein were located by using fluorescence microscope．Wholecell patch clamp technique was used to observe the effect of AVP on the firing frequency of MnPOVglut2 neurons，the effect of synaptic transmission blockers ( STBs) on the AVP-induced change in the firing frequency of MnPOVglut2 neurons，and the effect of AVP V1a receptor antagonist on the AVP-induced change in the firing frequency of MnPOVglut2 neurons. Results: The mean firing frequency of MnPOVglut2 neurons increased during perfusion with artificial cerebrospinal fluid (ACSF) and AVP compared with that during perfusion with ACSF (P＜0. 01) ，indicating that AVP excited the MnPOVglut2 neurons．The mean firing frequency of MnPOVglut2 neurons still increased during perfusion with ACSF，STBs，and AVP compared with that during perfusion with ACSF and STBs (P＜0. 001) ; moreover，the magnitude of AVP-induced increase in firing frequency didn't change significantly during perfusion with ACSF，STBs，and AVP compared with that during perfusion with ACSF and AVP (P ＞0. 05 ) ，suggesting that AVP excited the MnPOVglut2 neurons directly in a postsynaptic manner．The magnitude of AVP-induced increase in the firing frequency of MnPOVglut2 neurons declined during perfusion with ACSF，STBs，AVP，and V1areceptor antagonist compared with that during perfusion with ACSF，STBs，and AVP (P＜0. 01) ，suggesting that AVP excited MnPOVglut2 neurons directly via V1a receptor． Conclusion AVP can excite MnPOVglut2 neurons via V1areceptor directly in a postsynaptic manner．This study reveals the molecular marker of MnPO neurons which AVP act on.

Objective:To analyze the risk factors of prolonged mechanical ventilation (PMV) in patients with sepsis complicated by abdominal surgery, and to evaluate the predictive value of risk factors for PMV.Methods:A retrospective case-control study was conducted. The clinical data of patients with postoperative abdominal sepsis complicated with invasive mechanical ventilation who were admitted to the intensive care unit (ICU) of the Affiliated Hospital of Guizhou Medical University from January 1, 2018 to December 31, 2020 were collected. The patients were divided into PMV group (duration of mechanical ventilation longer than 48 hours) and non-PMV group (duration of mechanical ventilation shorter than 48 hours) according to the duration of mechanical ventilation in ICU. The patient's gender, age, body mass index (BMI), underlying diseases, mean arterial pressure (MAP), complete blood count, blood biochemistry, arterial blood gas, cardiac function indicators, procalcitonin (PCT) at admission to the ICU, the acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) and the sequential organ failure assessment (SOFA) scores in the first 24 hours of admission to the ICU, and other clinical information were recorded. Univariate and multivariate Logistic regression models were used to analyze the risk factors for PMV. Receiver operator characteristic curve (ROC curve) was drawn to evaluate the predictive value of related indicators for PMV.Results:A total of 195 patients with sepsis after abdominal surgery who received invasive mechanical ventilation were enrolled, including 127 males (65.1%) and 68 females (34.9%), with the median age of 65 (21, 93) years old. There were 91 patients (46.7%) in the non-PMV group and 104 patients (53.3%) in the PMV group. Univariate analysis showed that the APACHEⅡ score, SOFA score, cardiac troponin T (cTnT), N-terminal pro-B type natriuretic peptide (NT-proBNP) in the PMV group were significantly higher than those in the non-PMV group. Oxygenation index (PaO 2/FiO 2), total protein (TP) and prealbumin (PA) in the PMV group were all lower than those in the non-PMV group when admitted to ICU. In the PMV group, serum creatinine (SCr), blood urea nitrogen (BUN), cystatin C (Cys C) were significantly increased, prothrombin time (PT) was significantly prolonged, the proportion of patients with septic shock and hypertension were significantly increased as compared with those in the non-PMV group. Multivariate analysis showed that low PaO 2/FiO 2 at ICU admission [odds ratio ( OR) = 0.995, 95% confidence interval (95% CI) was 0.992-0.999, P = 0.010], high ln PCT ( OR = 1.301, 95% CI was 1.088-1.555, P = 0.004), high ln cTnT ( OR = 1.562, 95% CI was 1.079-2.261, P = 0.018) and septic shock ( OR = 4.967, 95% CI was 2.461-10.026, P = 0.000) were the independent risk factors for PMV in patients with sepsis after abdominal surgery. ROC curve analysis showed that the PaO 2/FiO 2, ln cTnT, ln PCT and septic shock had certain predictive value for PMV, the area under the ROC curve (AUC) of the four variables were 0.607, 0.638, 0.690 and 0.711, the sensitivity was 50.0%, 62.5%, 86.5% and 74.0%, and the specificity was 71.4%, 62.6%, 48.3% and 68.1%, respectively. The AUC for the joint prediction of the four variables was 0.803, with a sensitivity of 76.0% and a specificity of 78.0%. It suggested that the multivariate joint prediction of PMV was more accurate. Conclusions:Decreased PaO 2/FiO 2, increased PCT, increased cTnT and the occurrence of septic shock are independent risk factors for PMV in patients with sepsis complicated by abdominal surgery. The combination of above four indices was more accurate than one single variable in predicting PMV and had higher diagnostic value.

Objective:To observe the effects of berberine on procoagulant and fibrinolytic inhibitory factors produced by rat type Ⅱ alveolar epithelial cell (AECⅡ) induced by lipopolysaccharide (LPS).Methods:AECⅡ cells (RLE-6TN cells) were cultured in vitro, and the cells in logarithmic growth phase were collected. The cytotoxicity text of berberine was detected by cell counting kit-8 (CCK-8) to determine the drug concentration range according to inhibition concentration of half cells (IC 50). The RLE-6TN cells were divided into five groups, the cells in blank control group were cultured in DMEM; the cells in LPS group were stimulated with 5 mg/L LPS; and the cells in berberine pretreatment groups were pretreated with 20, 50 and 80 μmol/L berberine for 1 hour, and then were co-cultured with 5 mg/L LPS. The cells were collected after LPS induced for 24 hours. The protein and mRNA expression levels of tissue factor (TF), tissue factor pathway inhibitor (TFPI) and plasminogen activator inhibitor-1 (PAI-1) in the cells were detected by Western blotting and real-time fluorescence quantification reverse transcription-polymerase chain reaction (RT-qPCR). The levels of activated protein C (APC), precollagen Ⅲ peptide (PⅢP), thrombin-antithrombin complex (TAT) and antithrombin Ⅲ (ATⅢ) in the cell supernatant were measured by enzyme linked immunosorbent assay (ELISA). Results:According to the inhibition rate curve, the IC 50 of berberine on RLE-6TN cells was 81.16 μmol/L. Therefore, 20, 50 and 80 μmol/L were selected as the intervention concentration of berberine. Compared with the blank control group, the expression and secretion of procoagulant and fibrinolytic inhibitory factors were abnormal in RLE-6TN cells after LPS induced for 24 hours. The protein and mRNA expression levels of TF and PAI-1 in the LPS group were significantly increased, but the protein and mRNA expression levels of TFPI were significantly decreased. Meanwhile, the levels of APC and ATⅢ in the cell supernatant were significantly decreased, while the levels of PⅢP and TAT were significantly increased. After pretreatment with berberine, the abnormal expression and secretion of procoagulant and fibrinolytic inhibitory factors induced by LPS were corrected in a dose-dependent manner, especially in 80 μmol/L. Compared with the LPS group, the protein and mRNA expression levels of TF and PAI-1 in the berberine 80 μmol/L group were significantly decreased [TF protein (TF/GAPDH): 0.45±0.02 vs. 0.55±0.03, TF mRNA (2 -ΔΔCt): 0.39±0.08 vs. 1.48±0.11, PAI-1 protein (PAI-1/GAPDH): 0.37±0.02 vs. 0.64±0.04, PAI-1 mRNA (2 -ΔΔCt): 1.14±0.29 vs. 4.18±0.44, all P < 0.01] and those of TFPI were significantly increased [TFPI protein (TFPI/GAPDH): 0.53±0.02 vs. 0.45±0.02, TFPI mRNA (2 -ΔΔCt): 0.94±0.08 vs. 0.40±0.05, both P < 0.01]. Meanwhile, the levels of APC and ATⅢ in the cell supernatant were significantly increased [APC (μg/L): 1 358.5±26.0 vs. 994.2±23.1, ATⅢ (μg/L): 118.0±7.4 vs. 84.4±2.7, both P < 0.01], while those of PⅢP and TAT were significantly decreased [PⅢP (μg/L): 11.2±0.4 vs. 18.6±0.9, TAT (ng/L): 222.1±2.8 vs. 287.6±7.0, both P < 0.01]. Conclusions:Berberine could inhibit the LPS-induced expressions of procoagulant and fibrinolytic inhibitory factors in rat AECⅡ cells and promote the expressions of anticoagulant factors in a dose-dependent manner. Berberine may be a new therapeutic target for alveolar hypercoagulability and fibrinolysis inhibition in acute respiratory distress syndrome (ARDS).

Objective:To determine the effect of andrographolide (AD) on the expression of procoagulant and fibrinolytic inhibitory factors in rat type Ⅱ alveolar epithelial cells (AECⅡ) stimulated by lipopolysaccharide (LPS).Methods:The AECⅡ cells RLE-6TN in the logarithmic growth phase were divided into 5 groups: the normal control (NC) group, the LPS group, and the 6.25, 12.5, and 25 mg/L AD groups (AD 6.25 group, AD 12.5 group, AD 25 group). The NC group was cultured with RPMI 1640 conventional medium. In the LPS group, 5 mg/L LPS was added to the RPMI 1640 conventional medium for stimulation. Cells in the AD groups were treated with 6.25, 12.5, and 25 mg/L AD in advance for 1 hour and then given LPS to stimulate the culture. The cells and cell culture supernatant were collected 24 hours after LPS stimulation. The protein and mRNA expressions of tissue factor (TF), tissue factor pathway inhibitor (TFPI), and plasminogen activator inhibition-1 (PAI-1) in cells were detected by Western blotting and real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). The levels of procollagen Ⅲ peptide (PⅢP), thrombin-antithrombin complex (TAT), antithrombin Ⅲ (AT-Ⅲ) and activated protein C (APC) in the cell supernatant were detected by enzyme linked immunosorbent assay (ELISA).Results:Compared with the NC group, the protein and mRNA expressions of TF and PAI-1 in the LPS group were significantly increased, and the protein and mRNA expressions of TFPI were significantly reduced. At the same time, the levels of PⅢP and TAT in the cell supernatant were significantly increased, the levels of AT-Ⅲ, APC were significantly reduced. Compared with the LPS group, the protein and mRNA expressions of TF and PAI-1 in AD 6.25 group, AD 12.5 group, AD 25 group were significantly reduced [TF/GAPDH: 0.86±0.08, 0.45±0.04, 0.44±0.04 vs. 1.32±0.10, TF mRNA (2 -ΔΔCt): 2.59±0.25, 2.27±0.05, 1.95±0.04 vs. 4.60±0.26, PAI-1/GAPDH: 2.11±0.07, 1.45±0.04, 0.86±0.09 vs. 2.56±0.09, PAI-1 mRNA (2 -ΔΔCt): 3.50±0.22, 2.23±0.29, 1.84±0.09 vs. 6.60±0.27, all P < 0.05], while the protein and mRNA expressions of TFPI were significantly increased [TFPI/GAPDH: 0.78±0.05, 0.81±0.03, 0.84±0.07 vs. 0.36±0.02, TFPI mRNA (2 -ΔΔCt): 0.46±0.09, 0.69±0.07, 0.91±0.08 vs. 0.44±0.06, all P < 0.05]. Also the levels of PⅢP and TAT in the cell supernatant were significantly reduced, and the levels of AT-Ⅲ and APC were significantly increased [PⅢP (μg/L): 13.59±0.23, 12.66±0.23, 10.59±0.30 vs. 15.82±0.29, TAT (ng/L): 211.57±6.41, 205.69±4.04, 200.56±9.85 vs. 288.67±9.84, AT-Ⅲ (μg/L): 102.95±3.86, 123.92±2.63, 128.67±1.67 vs. 92.93±3.36, APC (μg/L): 1 188.95±14.99, 1 366.12±39.93, 1 451.15±29.69 vs. 1 145.55±21.07, all P < 0.05]. With the increase of the dose of AD, the above-mentioned promotion and inhibition effects became more obvious. In the AD 25 group, TF, PAI-1 protein and mRNA expressions decreased, TFPI mRNA expression increased, PⅢP level in the supernatant decreased and AT-Ⅲ, APC levels increased compared with AD 6.25 group, the difference was statistically significant, and the decrease of PAI-1 protein expression and PⅢP level in the supernatant were also statistically significant compared with AD 12.5 group. Conclusions:Andrographolide in the dose range of 6.25-25 mg/L can dose-dependently inhibit the expression and secretion of procoagulant and fibrinolytic inhibitor-related factors in AECⅡ cells RLE-6TN stimulated by LPS, and promote the secretion of anticoagulant factors. 25 mg/L has the most obvious effect.

Objective:To investigate the effect of NEMO binding domain peptide (NBDP) on lung inflammation and apoptosis in mice with acute respiratory distress syndrome (ARDS) and its mechanism.Methods:Thirty-six male BALB/c mice were divided into normal saline (NS) control group, ARDS model group, NBDP negative control group and 6, 12 and 18 μg NBDP pretreatment group by random number table method, with 6 mice in each group. ARDS mouse model was reproduced by aerosol inhalation lipopolysaccharide (LPS) 50 μL. An equivalent among of NS was inhaled in NS control group. The mice in NBDP negative control group were inhaled the materials similar to the non-functional NBDP 30 minutes before the aerosol inhalation LPS; 6, 12 and 18 μg of NBDP 50 μL were respectively inhaled in NBDP pretreatment groups. After inhalation of LPS for 6 hours, mice were sacrificed to get lung tissue and observe the degree of pathological injury and edema. Western blotting was used to detect the phosphorylation of nuclear factor-κB (NF-κB) pathway related proteins [NF-κB inhibitor (IκB) kinaseα/β(IKKα/β), IκBα and NF-κB p65; p-IKKα/β, p-IκBα, p-p65] and the expression of caspase-3 in lung tissue. The bronchoalveolar lavage fluid (BALF) was collected and the levels of inflammatory markers such as myeloperoxidase (MPO), interleukins (IL-1β, IL-8), and tumor necrosis factor-α (TNF-α) were detected by enzyme linked immunosorbent assay (ELISA).Results:ARDS model group had severe edema and hemorrhage, alveolar structure destruction, pulmonary hemorrhage and hyaline membrane formation etc. under light microscope, consistent with the pathological characteristics of ARDS lung tissue, suggesting that the ARDS model was successfully reproduced. ELISA showed that MPO, IL-1β, IL-8 and TNF-α levels of BALF in ARDS model group were obviously higher than those in NS control group. There were no significant differences in the above inflammatory indicators between NBDP negative control group and ARDS model group. The levels of MPO, IL-1β, IL-8 and TNF-α in NBDP pretreatment groups were significantly lower than those in ARDS model group in a dose-dependent manner, especially in 18 μg NBDP, the differences were statistically significant as compared with ARDS model group [MPO (ng/L): 393.32±19.35 vs. 985.87±101.50, IL-1β (ng/L): 43.05±5.11 vs. 97.68±10.88, IL-8 (ng/L): 84.64±2.32 vs. 204.00±17.37, TNF-α (ng/L): 229.13±17.03 vs. 546.73±62.72, all P < 0.05]. Western blotting showed that p-IKKα/β, p-IκBα, p-p65 and caspase-3 protein expressions in ARDS model group were significantly higher than those in NS control group. There was no significant difference in above NF-κB pathway and apoptosis-related protein expression between the NBDP negative control group and ARDS model group. The p-IKKα/β, p-IκBα, p-p65 and caspase-3 protein expression in NBDP pretreatment groups were significantly lower than those in ARDS model group in a dose-dependent manner, especially in 18 μg NBDP, the differences were statistically significant as compared with ARDS model group [p-IKKα/β protein (p-IKKα/β/β-actin): 0.15±0.02 vs. 0.42±0.04, p-IκBα protein (p-IκBα/β-actin): 0.10±0.01 vs. 0.93±0.30, p-p65 protein (p-p65/β-actin): 0.22±0.05 vs. 1.37±0.21, all P < 0.05]. Conclusion:NBDP can inhibit inflammatory response and apoptosis in ARDS lung tissue in a dose-dependent manner, and its mechanism is associated with interference NF-κB signaling pathway transduction.

The blood donation, component preparation and manufacturing, laboratory, issuing and quality control were studied and compared between the UK and China, in order to learn British experience in the clinical practice and blood services. The age limits of blood donors, blood collection units, donation times per year, laboratory items, and the types(volumes) of component preparation and manufacturing in the UK are more superior than those in China. In addition, the blood quality monitoring and regarding indicators are more scientific and reasonable in the UK. The full reimbursement of clinical blood expenses for patients has been realized in the UK. The British experience in continuous safeguard of the blood safety and balance of requirement and availability contributes to the constant and scientific development of British blood services over the years, and is worthy of references.