Objective: To investigate the regulatory effect of CLEC2D-CD161 interaction on killing capacity of decidual natural killer (dNK) cells during early pregnancy and its association with the incidence of recurrent spontaneous abortion (RSA). Methods: Decidua tissues were collected from normal pregnancies (n=16) and RSA cases (n=6) at 6-10 gestational weeks in the Department of Obstetrics and Gynecology of Peking University Third Hospital from October 2018 to May 2019. (1) Expressions of CLEC2D and CD161 in decidua from early pregnancy were detected using immunofluorescence. (2) Primary dNK cells were isolated from decidua from early pregnancy. dNK cells pre-treated with CD161 antibody (blocking CD161, B-CD161) were co-cultured with JEG-3 cells which were knocked-down by CLEC2D small interfering RNA (siCLEC2D), followed by killing capacity assessment of dNK cells by cytotoxicity assay and determination of expressions of related molecules by quantitive real-time polymerase chain reaction. (3) Western blot and flow cytometry were used to detect the expression of CLEC2D and CD161 in decidua tissues. Cytotoxicity assay was performed to analyze the killing capacity of dNK cells. T test was used for statistical analysis between normal and RSA cases. Results: (1) CLEC2D was mainly expressed in extravillous trophoblast (EVT) cells and CD161 was mainly detected in dNK cells. CD161-positive dNK cells and CLEC2D-positive EVT cells were adjacently located in decidua tissues allowing their interaction. (2) Cytotoxicity assay suggested that CD161 blocking in dNK cells or CLEC2D knockdown in JEG-3 cells could enhance the cytotoxicity of dNK cells. The target cell lysis rates at the effector-target ratios of 40∶1, 20∶1, 10∶1 and 5∶1 in B-CD161 group were (59.12±4.56)%, (25.96±5.44)%, (13.60±8.94)% and (12.53±8.94)%, and in IgG control group were (20.01±1.96)%, (8.51±1.32)%, (3.24±0.75)% and (3.82±1.92)%, respectively. There were significant differences between the two groups at the effector-target ratios of 40∶1 (t=13.922, P<0.01) and 20∶1 (t=5.403 P<0.05), but not at 10∶1 or 5∶1 (P>0.05). The target cell lysis rates at the effector-target ratios of 40∶1, 20∶1, 10∶1 and 5∶1 in si-CLEC2D group were (43.37±2.01)%, (32.99±2.08)%, (23.47±1.36)% and (11.48±0.37)%, and in the negative control (NC) group were (15.54±1.46)%, (13.84±1.68)%, (9.94±3.01) and (5.50±0.99)%, respectively. Differences between the two groups at all effector-target ratios were statistically significant (t=19.402, 12.400, 7.093 and 9.842, all P<0.01). Moreover, the expression of dNK killing-related factor granzyme B in the siCLEC2D group was higher than that in the NC group. (3) Compared with the normal pregnancy group, the RSA group showed decreased CD161 expression and increased killing capacity of dNK cells, but no significant difference in CLEC2D expression. Conclusions At early pregnancy, CLEC2D on EVT cells can interact with CD161 on dNK cells, which inhibits the cytotoxicity of dNK cells and induces immune tolerance at the fetal-maternal interface. Decreased expression of CD161 in decidua results in increased cytotoxicity of dNK cells, which may be one of the causes of immune rejection in RSA.

Objective To observe the effect of subarachnoid block with 0.5% bupivacaine at different temperatures during cesarean section.Methods 100 cases of elective cesarean section were randomly divided into room temperature group and heating group,50 cases in each group.Room temperature group: bupivacaine hydrochloride injection and glucose injection equilibrated group in a constant temperature thermostatic bath of 24 degrees thermostatic bath heating for above 30 min.Heating group: bupivacaine hydrochloride injection and glucose injection heated in the constant temperature thermostatic bath of 37 degrees thermostatic bath heatingfor above 30 min.Anesthesia was injected into the subarachnoid space at different temperatures to observe the anesthetic effect.Results The anesthesia increased rapidly, and the analgesia and muscle relaxation effects were better in the heating group than room temperature group, but the heating group had hypotension rate was higher than the room temperature group (36.0% vs.16.0%).There was no obvious difference between the incidence of adverse reactions such as nausea and vomiting in both groups.Conclusion Different temperatures of bupivacaine can be used safely for section anesthesia.The anesthesia effect of the heateding bupivacaine is faster, the anesthesia level is higher, the anesthesic and muscle relaxant effect is better.Bupivacaine at room temperature has relatively small effect on hemodynamics.

Objective To compare the effects of a single injection of palonosetron and tropise-tron to prevent postoperative nausea and vomiting (PONV ) in gynecological surgeries. Methods Sixty patients undergoing elective major gynecological surgeries with general anesthesia (Apfel score ≥ 3 ) were included and randomized to group P (palonosetron ) and group T (tropisetron),30 patients in each group.All patients received general anesthesia with tracheal intuba-tion,and palonosetron 0.25 mg or tropisetron 5 mg were injected before anesthesia respectively in two groups.Intravenous hydromorphine was delivered for postoperative analgesia in all patients. PONV were evaluated and followed up for 72 hours.The degree of PONV was recorded and the com-plete response rate (CR)and complete control rate (CC)were calculated.Results The degree of PONV in 0-24 h and 24-48 h was milder in group P than in group T (P <0.05),while in 48-72 h the degree of PONV was similar between the two groups.In group P,5 patients had vomiting postopera-tively with the failure period of treatment of (1 9.6±9.4)h,and no patients needed rescue treatments. While in group T,1 9 patients suffered from vomiting with the failure period of treatment of (20.6± 4.5)h,and rescue treatments were delivered 3 times in total.The CR and CC of preventing PONV in 0-24 h,24-48 h and 0-72 h were significantly higher in group P than in group T (P <0.05).The CR and CC of the two groups were comparable in 48-72 h postoperatively.Conclusion A preoperative single dose of palonosetron 0.25 mg is better than tropisetron 5 mg in preventing PONV within 48hours after gynecological surgery.

Anemia, Sideroblastic ; diagnosis ; genetics ; Asian Continental Ancestry Group ; Genetic Diseases, X-Linked ; diagnosis ; genetics ; Humans ; Pedigree

Objective To evaluate the effect of lithium chloride (LiCl) pretreatment on isoflurane-induced cognitive dysfunction and inflammatory response in hippocampus in aged rats.Methods Eighty 20-month-old male Sprague-Dawley rats,weighing 350-400 g,were randomly assigned into 4 groups (n =20 each):control group (group C),1.4％ isoflurane group (group I),100 mg/kg LiCI + 1.4％ isoflurane group (group L+ I),and 100 mg/kg LiC1 group (group L).Group I was exposed to 1.4％ isoflurane in 30％ O2-70％ N2 for 6 h,while group C was exposed to 30％ O2-70％ N2 only.LiCl 100 mg/kg was injected intraperitoneally once a day for 3 consecutive days and isoflurane anesthesia was performed on 4th day in group L + I.LiCl 100 mg/kg was injected intraperitoneally once a day for 3 consecutive days and then the rats inhaled 30％ O2-70％ N2 for 6 h on 4th day in group L.Blood samples were taken immediately after the end of anesthesia for blood gas analysis.Hippocampi were isolated 24 h after the end of anesthesia for determination of the expression of glycogen synthase kinase-3β (GSK-3β) and acetyl-NF-κB (Lys310) (by Western blot) and TNF-α,IL-1β and IL-6 mRNA (by RT-PCR).The levels of TNF-α,IL-1β and IL-6 were determined by ELISA and the contents of TNF-α,IL-1β and IL-6 were calculated.The cognitive function was assessed on 2nd day after the end of anesthesia.Results Compared with group C,the expression of GSK-3β and acetyl-NF-κB (Lys310) was significantly up-regulated,the expression of TNF-α,IL-1β and IL-6 mRNA and contents of TNF-α,IL-1β and IL-6 were increased,the escape latency was prolonged,and the time of staying at the original platform quadrant was shortened in group I (P ＜ 0.05).Compared with group I,the expression of GSK-3β and acetyl-NF-κB (Lys310) was significantly down-regulated,the expression of TNF-α,IL-1β and IL-6 mRNA and contents of TNF-α,IL-1β and IL-6 were decreased,the escape latency was shortened,and the time of staying at the original platform quadrant was prolonged in group L + I (P ＜ 0.05).Conclusion LiC1 pretreatment can improve isoflurane-induced cognitive dysfunction and inhibition of inflammatory response in hippocampus is involved in the mechanism in aged rats.

ObjectiveTo observe the effect of penehyclidine hydrochloride (PHC) on serum interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α ) concentrations following tourniquet deflation in patients undergoing lower limb surgery.Methods Thirty adult patients,scheduled for unilateral lower limb surgery,ASA classification [ - Ⅱ grades,were divided into control group and research group by random number table,each group of 15 cases.Before anesthesia 30 min,PHC in intravenous infusion of 0.01-0.02 mg/kg in research group,the corresponding volume in intravenous infusion of 0.9% sodium chloride in control group.Peripheral venous blood samples were collected immediately before tourniquet inflation (T0,baseline),immediately before tourniquet deflation(T1),30 min (T2) and 60 min (T3) after tourniquet deflation.Serum IL-6 and TNF- α concentrations were measured by enzyme linked immunosorbentassay.ResaltsSerum TNF- α change at the same time point after tourniquet deflation was not statistically significant between two groups (P ＞ 0.05).Serum IL-6 concentration was decreased at each time point after tourniquet deflation compared with T0 in research group,while increased in control group.Serum IL-6concentration difference of T3 and T0 had statistically significant between research group and control group [ (-8.8 ± 5.6) ng/L vs.( 10.2 ± 6.7) ng/L,P＜ 0.05].ConclusionsPHC in advance can decrease serum IL-6 concentration after tourniquet deflation in patients undergoing lower limb surgery.

ObjectiveTo study the relationship of the parental rearing behaviors between the psychological status and post-traumatic stress disorder in the injured deliberately.Methods 161 injured people were treated in five hospitals of the Urumqi and evaluated the self-reporting questionnaire-20 (SRQ20),7-items screening scale for PTSD( PTSD7 ),impact of event scale (IES) and symptom checklist 90 (SCL-90) in the third and the fourth week injured,including 113 injured people aged at 14 ～50 years old completed the Egna Minnen av Barndoms Uppfostran (EMBU).Two months after the injury,113 injured people were interviewed,including 106 injured people diagnosed by Structured Clinical Interview for DSM-Ⅳ-TR Axis Ⅰ Disorders/Patients (SCID-I/P) and 7 injured people were lost.ResultsFather's preferences correlated with the total score of IES ( r =- 0.234,P ＜ 0.05 ) and avoid factor( r=-0.309,P＜ 0.01 ) positively,Mother's emotional warmth and understand correlated with the flashback factor of IES ( r =0.194,P ＜ 0.05 ) negatively.The injured people were divided into three groups by diagnosis:the illness-free group,the PTSD group,and other group.Parental preferences correlated with Mental symptoms negatively.Father's punish severely,interference too much,refuse or deny and Mother's interference too much,over protection and punish severely correlated mental symptoms positively.The six subscales of father's rearing behaviors and the five subscales of mother's rearing behaviors had no significant difference.ConclusionsParental preferences in childhood can internalizes inside support to protect the psychological trauma in future.Father's punish severely,interference too much,refuse or deny and mother's interference too much,over protection and punish severely can damage mental health and aggravate symptoms after traumatic event.But the influence of parental rearing behaviors is limited to decide whether the injured people suffering from PTSD or other mental disorders.

Objective To investigate the effect of melatonin on ketamine-induced apoptosis in hippocampal neurons of fetal rats.Methods Sixteen to eighteen day pregnant Sprague Dawley rats were anesthetized.The fetal rats were obtained under sterile condition and decapitated.The hippocampal neurons were isolated and primary cultured for 5 days.The primary cultured neurons were randomly divided into 5 groups (n =6 each):control group (group C),ketamine group (group K),and 1.0,2.5 and 5.0 mmol/L melatonin groups (groups M1-3 respectively).Ketamine with the final concentration of 1 000 μmol/L was added to the culture medium and the neurons were incubated for 3 h in group K.In groups M1-3,1.0,2.5 and 5.0 mmol/L melatonin were added to the culture medium,respectively,at 60 min before the addition of ketamine,and the neurons were incubated for 3 h.While the equal volume of normal saline was added instead in group C.The neuronal viability during the developmental phase was assessed by MTT assay.The mitochondrial membrane potential (Ψm) was measured by flow cytometry.The expression of cAMP response element binding protein phosphorylation (p-CREB (Ser133)),Bcl-2,Bax,and cytochrome C was detected by Western blot.Results Compared with group C,the neuronal viability and Ψm were significantly decreased,and the expression of p-CREB and Bcl-2 was down-regulated,while the expression of Bax and cytochrome C was up-regulated in group K (P ＜ 0.05).Compared with group K,Ψm was significantly increased in groups M2 and M3,and the neuronal viability was significantly increased,the expression of Bcl-2 was up-regulated,while the expression of Bax and cytochrome C was down-regulated in groups M1-3 (P ＜ 0.05).Conclusion Melatonin can protect the hippocampal neurons of fetal rats from apoptosis triggered by ketamine via regulating the expression of Bcl-2 and Bax,stabilizing Ψm,inhibiting the release of cytochrome C from mitoehondria,and preventing apoptosome formation.

ObjectiveTo summarize and analyse the perioperative management especially theanesthesia of 20 kidney transplant recipients in non-transplant surgery. MethodThe anesthesia management of 20 kidney transplant recipients in non-transplant surgery was analyzed retrospectively. ResultsIn 20 cases, 1 case (5％) was performed under local anesthesia,4 cases (20％) were performed under intravertebral anesthesia and 15 cases (75％) were performed under general anesthesia. The operation time was 30-260 min, all cases were managed successfully. ConclusionIt is still a clinical challenge to deal with the surgical patients after kidney transplantation, and it needs fully understanding of the pathophysiological status of the patient and closely collaboration of transplant physicians, anesthesiologists and the surgeons.

Objective To identify the clinical features, the molecular diagnosis and the molecular mechanism of three unrelated factor X deficiency families. Methods Three probands were male and the diagnosis was validated by coagulant parameters. The F X coagulation activity ( F X∶ C ) and antigen (FX∶ Ag) were tested by clotting test and ELISA method. The cross-corrected test was used to rule out the inhibitor of FX in plasma. Thrombin generation test was evaluated. The antigen and the molecule weight of the FX in plasma were measured with western blotting. Gene mutations were analyzed in the probands and their family members with PCR and DNA sequencing. FX expression plasmids were constructed and transientby being transfected into 293T cells. FX: C and FX: Ag of the expression products were tested. Results APTT and PT in proband 1 were obviously prolonged, 113.4 s and 62.3 s, respectively. And there was no inhibitor in plasma. The thrombin generation was lower compared to normal reference. APTT and PT in proband 2 were 56. 5 s and 28.7 s. There was no inhibitor in the plasma. The thrombin generation was 1 101.5 nmol · min. APTT and PT in proband 3 were 117.3 s and 44. 3 s. The thrombin generation was 782.5 nmol · min. FX∶ C and FX∶ Ag in proband 1 were 1.4% and 3.6%, with a homozygous mutation in FX gene (Ser425→Pro). In vitro expression of the mutation showed a normal synthesis in the cell but secretion dysfuntion. In proband 2 F X: C and F X: Ag were 2. 2% and 5. 5%, with two heterozygous mutations in FX gene (Ala-29→Pro and Phe324→Leu). The Ala-29 → Pro mutation led to significantly reduced expressions of FX in both cell lysate and cell culture supernatants compared to wild-type plasmid,(41.32 ±5.21 )% and(6. 30 ± 1.84)% respectively. However Phe324→Leu mutation almost did not affect the FX synthesis. FX: C and FX: Ag in proband 3 were 2. 2% and 35%, with two heterozygous mutations in FX gene( Ala235→Thr and Arg347→Cys). The expressions of these two mutant FX proteins in cell lysate were similar to those of wild-type but obviously lower in the supernatant. Conclusions Five mutations of F X gene are found in this study. These mutations (Ser425Pro, Phe324Leu, Ala235Thr and Arg347Cys)can not affect F X protein synthesis. However Ala-29Pro mutation can reduce F X protein synthesis and cause secretion dysfunction.