The cerebral resuscitation is the most important aim in advanced cardiopulmonary cerebral resuscitation (CPCR). Cerebral function protection after cardiac arrest (CA) is important to improve survival rates including those after the discharge. Therapeutic mild hypothermia maybe the only method that can improve neurological function of patients following resuscitation after CA, which was recommended as one of treatment strategies for unconscious patients after successful resuscitation in 2010 American Heart Association Guidelines for Cardiopulmonary Resuscitation and Emergency Cardiovascular Care. Although there are many cooling methods of inducing hypothermia, each has some advantages or shortcomings. There were many controversies on the choice, which need further research. We make a summary and analysis about the mechanism of therapeutic mild hypothermia, the possible complications at different stages and the clinical application of mild hypothermia, such as the evaluation before therapeutic mild hypothermia and the choice of hypothermia protocol and cooling methods, to provide guidance for clinical mild hypothermia therapy.

Adolescent ; Anemia, Aplastic ; complications ; therapy ; Anemia, Hemolytic ; chemically induced ; complications ; Antilymphocyte Serum ; adverse effects ; therapeutic use ; Female ; Humans

BACKGROUND:Methicil in-resistant Staphylococcus aureus has been a primary pathogen of nosocomial infections worldwide. To construct a quick and easy knockout method is an important technique of studying virulence and resistance of methicil in-resistant Staphylococcus aureus. OBJECTIVE:To construct the Staphylococcus aureus gene knockout plasmid for understanding the antibiotic resistance and virulence of Staphylococcus aureus. METHODS:pUC19 was considered as a basic skeleton of construction. pLE194Ts temperature-sensitive replicon and tetracycline resistance gene fragment pHY300PLK plasmid in pCL52.1 were bound to EcoR I site in pUC19 by high assurance amplification. Al multiple clone sites in pUC19 were reserved. The Escherichia coli-Staphylococcus aureus shuttle plasmid was obtained. The N315 dapB gene knockout plasmid was obtained through gene knockout technology. This strain was eventual y identified by multiplex-PCR. RESULTS AND CONCLUSION:The Escherichia coli-Staphylococcus aureus shuttle plasmid, pYZ1 and pYZ8, was successful y constructed, and had been used in Staphylococcus aureus gene knockout. Homologous recombinant plasmid pYZ-ΔdapB was constructed by restriction enzyme digestion and overlap technique. After genetical y modification in RN4220, the constructed gene knockout plasmid pYZ-ΔdapB was introduced to N315 to be screened in the low culture temperature. The deletion strain was successful y obtained after being identified by multiplex-PCR. Above data suggested that pYZ1 and pYZ8 can be successful y used for Staphylococcus aureus gene detection, which provides a tool to study resistance and virulence of clinical Staphylococcus aureus strains.

Objective To investigate the clinical outcome and placental characteristics of spontaneous twin anemia-polycythemia sequence (sTAPS). Methods Twelve cases with sTAPS delivered in Peking University Third Hospital from May 2013 to August 2016. The data of ultrasound characteristics, gestational age at delivery, and 1 minute Apgar score were analyzed,retrospectively. Placental superficial vascular anastomoses, placental territory discordance and the ratio of umbilical cords insertion distance to the longest placental diameter were also analyzed. Results (1) Only 1 case of sTAPS was diagnosed prenatally, the others were diagnosed postnatally because the fetal middle cerebral artery(MCA) doppler was not measured regularly. Five cases were complicated with selective intrauterine growth restriction (sIUGR). The median gestational age at delivery was 32.8 weeks(31-37 weeks). The pregnancies were terminated because 3 cases were sIUGR typeⅠ, 1 case was sIUGR type Ⅱ, 1 case was sIUGR type Ⅲ, 2 cases were fetal distress, 2 cases were severe pre-eclampsia, 2 cases were premature rupture of membrane, 1 case was fetal hydrops with abnormal doppler waveforms of ductus venouses. (2) When 5 sIUGR cases were excluded, there was no difference between the twins in birth weight [1797 g(940-2620 g),1648 g(980-2500 g);P=0.688]. The hemoglobin (Hb) level in all donor was significantly lower than recipient(P=0.000)and the inter-twin Hb difference was 147.6 g/L (84.0-216.0 g/L). While the reticulocyte percentage in donor was significantly higher than recipient (P=0.013) and reticulocyte percentage ratio was 3.60 (1.04-7.50). Five donor newborns had neonatal asphyxia, including 1 severe asphyxia, while no asphyxia happened in the recipient twins. (3) Arterio-arterial (A-A) anastomoses, veno-venous (V-V) anastomoses, arterio-venous (A-V) anastomoses were found in 3, 1 and 11 placentas, respectively. The total number of anastomoses was 2 (1-5) and the total diameter was 1.1 mm (0.4-2.1 mm), including 0 (0-1) A-A anastomoses with 0.2 mm (0.0-0.9 mm) in diameter and 2 (0-5) A-V anastomoses with 0.7 mm (0.0-2.1 mm) in diameter. The placental territory discordance was 0.17 (0.02-0.40) and the ratio of umbilical cords insertion to the longest placental diameter was 0.82 (0.34-0.99). Conclusions The pathogenesis of sTAPS might result from slow and chronic blood transfusion from donor to recipient through a few minuscule vascular anastomoses in the placenta. In all monochorionic twins, especially sIUGR cases, MCA doppler should be monitored closely in the second and third trimester, in order to diagnose and manage sTAPS in time.

Objective To evaluate the efficacy of fetoscopic laser occlusion of chorioangiopagous vessels (FLOC) in treatment of twin-twin transfusion syndrome (TTTS) and to investigate the incidence of TTTS complicated with selective intrauterine growth restriction (sIUGR) for better understanding the effects of sIUGR as a complication of TTTS on pregnancy outcomes. Methods A retrospective study was performed on 116 gravidas who were diagnosed with TTTS of Quintero stage Ⅰ - Ⅳ in Peking University Third Hospital from September 2008 to September 2014. Among them, 44 cases received FLOC therapy. The incidences of sIUGR in each Quintero stage of TTTS were analyzed. Pregnancy outcomes of those 44 cases treated with FLOC were observed and the differences among four stage groups were analyzed. Chi-square or Fisher exact test was performed for statistical analysis. Results (1) The survival rates of both twins from stage Ⅰ to Ⅳ groups were 4/7, 10/14, 5/19 and 3/4, respectively, with statistically significant difference (χ2=7.840, P=0.038), but that in stage Ⅲ group was lower than in stage Ⅱ group without significant difference (P'=0.008). Differences in the total fetal survival rate among the four groups were statistically significant [8/14, 75% (21/28), 32% (12/38) and 6/8, χ2=14.016, P=0.002]. The total fetal survival rate in stage Ⅲ group was significantly lower than that in stageⅡ group (P'<0.008). In patients with stage Ⅲ TTTS, those complicated with sIUGR, after receiving FLOC therapy, showed a lower total fetal survival rate than those without sIUGR [21% (6/28) vs 6/10, P<0.05]. (2) Among the 116 TTTS patients, there were 63 cases (54%) with sIUGR complication. Patients with sIUGR complication in TTTS stages Ⅰ to Ⅳ groups accounted for 40% (14/35), 48% (11/23), 78% (28/36) and 46% (10/22), respectively, and the differences were significant (χ2=11.963,P=0.007). The incidence of sIUGR in stage Ⅲ group was greater than that in stage Ⅰ group (χ2=10.482, P'=0.002), and those in both stage Ⅲ and Ⅱ groups were higher than in stage Ⅰgroup without significant difference. Conclusions TTTS patients in stage Ⅲ have lower survival rate of both twins and total fetal survival rates after FLOC therapy, which may be related to a higher incidence of concurrent sIUGR.

Objective To investigate the in vitro effect of sirolimus on the apoptosis of a cutaneous squamous cell carcinoma cell line, Colo-16 cells. Methods Cultured Colo-16 cells were treated with different concentrations (50, 100, 150, 200 nmol/L) of sirolimus for various durations ( 12, 24, 48, 72 hours). Subse-quently, cell proliferation was detected by MTT assay, and cell apoptosis by Annexin V-FITC and PI double staining. Morphological changes of the cells were observed with Hoechst 33258 fluorescent staining. Total RNA was extracted from Colo-16 cells treated with sirolimus for 48 hours, and subjected to reverse tran-scription (RT)-PCR for the detection of mRNA expression of B cell lymphoma/leukmia-2 (Bcl-2) and Bcl-2-associated X Protein (Bax). Results Sirolimus inhibited the proliferation of Colo-16 cells in a time-and dose-dependent fashion. The early apoptosis rate was 7.26%±0.26%, 8.34% ±0.19%, 9.86%±0.14%, 11.92% ±0.15% in Colo-16 cells treated with sirolimus of 50, 100, 150, and 200 nmol/L, respectively, signifi-candy higher than that in untreated cells (1.53%±0.09%, P < 0.05); a positive correlation was observed between the apoptosis rate and concentrations of sirolimus (r = 0.955, P = 0.000). Typical morphological changes of apoptosis, such as chromatin condensation and margination as well as nuclear fragmentation were observed by fluorescence staining. After treatment with sirolimus for 48 hours, a significant decrease was observed in the mRNA expression of Bcl-2, while an increase in that of Bax was noticed. Conclusion Sirolimus could induce Colo-16 cells apoptosis in vitro, which may be associated with the modulation of expression of apoptosis-regnlating genes, such as Bcl-2 and Bax.

0bjective To determine the effect of 2 therapies(early realignment versus delayed open urethroplasty)for posterior urethral injuries associated with pelvic fractures. Methods A sys-tematic review of all the relevant clinical trials was performed.Trials were identified from specialized trials register of Cochrane UGDP Group,the Coehrane library,additional electronic search(mainly MEDLINE、EMBSAE、CBM)and manual searching.Clinical trials comparing early realignment and delayed open urethroplasty of posterior urethral disruption associated with pelvic fractures were includ-ed.Statistics analysis was managed by using Review Manager 4.2. Results Ten clinical trials inclu-ding 470 patients were included.Meta-analysis indicated that odds ratio(OR)of the rate of urethral stricture was 0.27[95%confidence interval(95%CI)0.08,0.86,P=0.03].The OR of the percent-age of strictures needing urethroplasty was 0.25,95%CI 0.08,0.74,P=0.01. Conclusions Early realignment may provide better outcomes than delayed open urethroplasty after posterior urethral dis-ruption.It can decrease urethral stricture formation and the percentage of strictures needing urethro-plasty.But the results might be influenced by the bias of grouping,especially the severity of injuries.

Objective To establish the reference interva1 for venous blood leukocyte count and classification among children from 1 month to 9 years old in the laboratory .Methods 2 213 health children aged from 1 month to 9 years old were retrospectively analyzed from October 2012 to March 2015 .White blood cells counts were analyzed statistically in 4 groups according to age ,classi‐fication was in 7 groups .The results were obtained by Sysmex series fully automatic blood analyzer .Results White blood cells de‐crease showed a trend of increase with age and fell to adult basic level after 7 years old .The changes of lymphocytes and neutrophils proportion along with ag were most obvious and had a certain regularity .Except for 3- <7 years‐old group ,there was no influence of gender factor in leukocyte count and classification .Conclusion The regional children′s white blood cells count and classification of the lab reference range were obtained which could improve the diagnosis .

Objective:To investigate the impact of recombinant flagellin targeting TLR5 and NLRC4 simultaneously or respectively on innate immune cells in mice. Methods: Induction,expression,purification and identification of recombiant FliC,which were FliC(activating both TLR5 and NLRC4);FliCΔ90-97(unable to activate TLR5),FliC-L3A(unable to activate NLRC4),FliCΔ90-97:L3A(unable to activate both TLR5 and NLRC4). The mice were divided into five groups,namely group FliC,FliC-L3A,FliCΔ90-97,FliCΔ90-97:L3A and PBS,which were injected with 100μl PBS or 10μg recombinant flagellin intraperitoneally,three mice in each group. 12 h later,the mice were executed using dislocation of cervical vertebra and the splenic and peritoneal cells were isolated. The spleen was grinded into single-cell suspension. The proportion of neutrophils,NK cells,DCs and the expression level of CD80 and CD86 on DCs were evaluated with flow cytometry. Results:Group FliC,group FliC-L3A and group FliCΔ90-97 shared the similar proportion of neutrophils in peritoneal cavity ( P>0. 05 ) , and all of which were significantly higher than group PBS and group FliCΔ90-97 ( P<0. 01),and NK cells also showed the similar trend. Compared with group FliCΔ90-97 and FliCΔ90-97:L3A,the mean fluorescence intensities(MFIs) of CD80 and CD86 in group FliC and FliC-L3A increased significantly(P<0. 01). The proportion of Treg in spleen was highest among all groups. Conclusion:Activation of TLR5 and NLRC4 had similar chemotaxis of neutrophils and NK cells. The ex-pression of CD80 and CD86 on DCs were upregulated after stimulation by flagellin and TLR5-dependent. Activation of TLR5,but not NLRC4,increased the proportion of Treg in spleen.

Blood Gas Analysis ; Female ; Humans ; Infant, Newborn ; Infant, Newborn, Diseases ; therapy ; Intensive Care Units, Neonatal ; Male ; Respiration, Artificial