OBJECTIVE:To establish a method for simultaneous residual determination of dichloromethane and ethyl acetate in bisacodyl raw material. METHODS:Head-space GC was performed on the capillary column of 6% cyanopropyl phenyl-94% di-methyl polysiloxane(DB-624)by temperature programming,the temperature of injector was 220 ℃,detector was flame ionization detector with temperature of 250 ℃,carrier gas was high purity nitrogen with the flow rate of 3.0 ml/min,split ration was 1∶10, headspace heating temperature was 70 ℃,equilibration time was 30 min,volume of headspace vial was 5 ml,and the injection volume was 1 ml. RESULTS:The linear range was 6-120μg/ml for dichloromethane(r=0.999 9)and 50-1 000μg/ml for ethyl ac-etate(r=0.999 9);the limit of quantitation was 0.2,1.7 μg,limit of detection was 0.06,0.5 μg;RSDs of precision,stability and reproducibility tests were no higher than 3%;recoveries were 100.30%-102.00%(RSD=0.63%,n=9) and 100.10 %-101.30%(RSD=0.44%,n=9). CONCLUSIONS:The method is simple and accurate,and can be used for the simultaneous residual deter-mination of dichloromethane and ethyl acetate in bisacodyl raw material.

Objective To explore the effect of salubrinal (sal,an endoplasmic reticulum stress inhibitor) on radiosensitivity of human head and neck squamous carcinoma cells (HNSCC).Methods Cells were divided into two groups of sal treatment and its control.For drug treatment group,cells were treated with 10 mmol/L sal for different time (12,24,36 h) and then irradiated.The levels of a core protein GRP78 of endoplasmic reticulum stress (ERS) in HNSCC (KB,Fadu,and Detroit 562 cells)were analyzed by Western blot assay at different time (0,20 min,1 h,3 h,6 h,12 h,24 h and 48 h) after irradiation.Cell survival was measured with colony formation assay.Results Western blot assay revealed that the protein levels of GRP78 in three kinds of HNSCC significantly increased from 20 min to 1 h and peaked at 3 h after radiation (t =12.72,13.37,18.31,P ＜ 0.05).Compared with the control group,treatment of cells with sal decreased GRP78 protein levels (t =14.25,5.34,3.12,P ＜ 0.05) in three cell lines and also significantly enhanced radiation damage and reduced cell viability.The sensitization enhancement ratios (SER) of sal in three cell lines were 1.16,1.05 and 1.06,respectively.Conclusions Rradiosensitivity of HNSCC could be effectively enhanced by sal treatment.

Objective To explore the effects of quality improvement in delivery room resuscitation on very/extremely low birth-weight infants (VLBWI/ELBWI). Methods A retrospective analysis was performed to analyze the clinical data of VLBWI/ELBWI who were admitted to the Neonatal Intensive Care Unit (NICU) of Nanjing Maternity Hospital Affiliated to Nanjing Medical University from January to December 2015 (pre-improvement group, n=176) and of those who were admitted from January to December 2016 after the implementation of quality improvement program on delivery room resuscitation (post-improvement group, n=199). Several parameters were monitored, including resuscitation modalities [continuous positive airway pressure (CPAP) , peak inspiratory pressure (PIP)+positive end expiratory pressure (PEEP) with T-piece resuscitator and intubation rate in delivery room], neonatal body temperature and pH on NICU admission, respiratory outcomes, morbidity from intraventricular hemorrhage, necrotizing enterocolitis, retinopathy ofprematurity and hospitalization. Chi-square (or Fisher's exact test), t or rank Sum test was used for statistical analysis. Results There was no significant difference in gestational age, birth weight, gender proportion, delivery mode and Apgar scores between the two groups (all P>0.05). After implementing the quality improvement program, there was an increased overall usage of CPAP [85.9% (171/199) vs 66.3% (112/176), χ2=19.881, P<0.01] and PIP+PEEP with T-piece resuscitator [33.8% (67/199) vs 10.8% (12/176), χ2=19.819, P<0.01], but a decreased usage of balloon catheter ventilation [6.0% (12/199) vs 39.3% (44/176), χ2=53.682, P<0.01]. No significant change in intubation rate was observed(P>0.05). The average admission temperature increased after launching the quality improvement program [M (P25-P75), 36.2 (35.8-36.5) vs 35.6 (35.4-35.7)℃ , Z= - 9.681, P<0.01]. The morbidities of pulmonary hemorrhage within one week after birth [1.5% (3/199) vs 5.1% (9/176),χ2=3.921] and grade Ⅲ / Ⅳ intraventricular hemorrhage [1.1% (2/199) vs 11.9% (21/176), χ2=33.885] decreased along with the improvement in delivery room resuscitation (both P<0.05). The duration of invasive ventilation decreased as well [3 (1-6) vs 4 (2-9) d, Z= - 2.286, P<0.05]. Conclusions Quality improvement in delivery room resuscitation measures standardizes the management of delivery room resuscitation and improves the clinical outcomes of VLBWI/ELBWI.

Objective To evaluate the effect of dexmedetomidine on the quality of intraoperative wake-up test in the patients undergoing balloon occlusion test of the internal carotid artery.Methods Forty-two patients of either sex with intracranial aneurysm,aged 57-78 yr,weighing 53-86 kg,of American Society of Anesthesiologists physical status Ⅱ or Ⅲ,scheduled for elective balloon occlusion test of the internal carotid artery under general anesthesia,were assigned into 2 groups (n =21 each) using a random number table:propofol conbined with remifentanil group (group PR) and dexmedetomidine combined with propofol and remifentanil group (group DPR).In group DPR,dexmedetomidine was intravenously infused over 15 min in a loading dose of 0.5 μg · kg-1 before induction of anesthesia,followed by an infusion of 0.3 μg · kg-1 · h-1 throughout surgery.Propofol and remifentanil were given by target-controlled infusion (TCI) after infusion of the loading dose.The patients were mechanically ventilated after placement of the laryngeal mask airway.Maintenance of anesthesia was as follows:propofol and remifentanil were given by TCI with the target plasma concentrations of 0.5-1.0 μg/ml and 1-3 ng/ml,respectively,in group DPR;propofol and remifentanil were given by TCI with the target plasma concentrations of 3-5 μg/ml and 3-6 ng/ml,respectively,in group PR.Bispectral index (BIS) value was maintained at 40-60.Before wakeup test,propofol infusion was stopped and the target plasma concentration of remifentanil was decreased to 0.5 ng/ml in two groups,and the infusion rate of dexmedetomidine was decreased to 0.1 pg · kg-1 · h-1 in group DPR.The wake-up time was recorded and the wake-up quality was assessed.After admission to the operating room (T0,baseline),at 10 min before wake-up test (T1),immediately after patients were wakened (T2),at 10 min after patients were wakened (T3) and at the end of wake-up test (T4),the mean blood pressure (MAP),heart rate,respiratory rate (RR),SpO2 and BIS values were recorded.The development of intraoperative awareness,emergence time,postoperative agitation,nausea and vomiting,regurgitation and aspiration and severe pain was recorded.Results MAP,heart rate,SpO2 and RR were all within the normal range during wake-up period in two groups.Compared with the baseline at To,MAP was significantly decreased at Ti,3,4 in group PR,and BIS value was decreased at T1-4 in DPR and PR groups (P＜0.05).Compared with group PR,MAP was significantly increased at T1.3,BIS value was decreased at T24,the wake-up time was shortened,Ramsay sedation score and wake-up quality were increased,the emergence time was shortened,and the incidence of agitation was deceased (P＜0.05),and no significant change was found in verbal rating scale scores assessed after extubation in group DPR (P ＞ 0.05).No cardiovascular events,respiratory depression,intraoperative awareness,postoperative nausea and voniting,regurgitation and aspiration or severe pain was found in two groups.Conclusion Dexmedetomidine can raise the quality of intraoperative wake-up test in the patients undergoing balloon occlusion test of the internal carotid artery.

Objective To summarize the clinical features of graft failure (GF)after non-T-cell depleted haploidentical hematopoietic stem cell transplantation (Haplo-HCT), and to investigate the causes and treatment. Methods A retrospective analysis was carried out on 174 patientswho accepted the non-T-cell depleted Haplo-HCT from Jan 2012 to Dec 2013. The patients′ donor specific anti human leukocyte antigen antibodies (DSA) from the peripheral blood serum were detected and those DSA positive patients were treated by immunoglobulin or plasma exchange before transplatation. Results A total of three patients with acute myeloid leukemia got GF, the incidence rate was 1.72%. The patient with primary GF was given a secondHaplo-HCT, but did not get implanted with leukemia remission and three lineages persistently low , he was died of pulmonary infection eight monthes after the second transplant. One of the secondary GF patients was given peripheral blood mononuclear cells(PBMNCs) mobilized by granulocyte colony stimulating factor (G-CSF) from the donor, and got full donor chimerism on day 16 after infusion. The disease-free survival has been for 18 months. The other case was found that DSA was positive, the mean fluorescence intensity (MFI) value was 15000, then Rituximab and PBMNCs mobilized by G-CSF were administrated successively. On day 14 after infusion the partient got full donor chimerism , and MFI turned negative. The patient has been disease-free survival for 41 months. Conclusion Graft failure is a rare but fatal complication after non-T-cell depletedHaplo-HCT, Rituximab followed by PBMNCs are effective measures for DSA related GF, as were worthy of further study.

OBJECTIVETo investigate the influence of therapeutic bloodletting at Jing-well points and hypothermia on acute cerebral edema after traumatic brain injury (TBI) in rats.

METHODSSeventy-five SD rats were randomly divided into sham-operation group (Sham), TBI group (TBI), bloodletting group (BL), mild-induced hypothermia group (MIH), and bloodletting plus MIH group (BL + MIH) (n = 15). The model of TBI was established by electric controlled cortical impactor (eCCI). The rats of BL group were bloodletting at Jing-well points immediately after injury, twice daily. While the MIH group was settled on a hypothermia blanket promptly after TBI for 6 hours, so that the temperature dropped to 32 degrees. Each of measurement was performed after 48 hours. Magnetic resonance imaging (MRI) was used to evaluate the dynamic impairment of cerebral edema after TBI (n = 3). In addition, mNSS score, measurements of wet and dry brain weight, and Evans Blue assay were performed to investigate the neurologic deficit, cerebral water content (n = 8), and blood-brain barrier permeability (BBB), (n = 4), respectively.

RESULTSMRI analysis showed that the cerebral edema, hematoma and midline shifting of rats in TBI group was more serious than other treatment group. Meanwhile compared with TBI group, the mNSS scores of every treatment group were meaningfully lower (all P < 0.05). Furthermore, treatment with BL+ MIH group was superior to the separated BL and MIH group (all P < 0.01). In addition, brain water content of each intervention group reduced to varying degrees (all P < 0.05), especially that of MIH group and BL + MIH group (P <0.01). BBB permeability of each treatment group was also significantly improved (all P < 0.01), and the improvement in MIH group and BL + MIH group was much better than the BL alone group (P < 0.05, P < 0.01).

CONCLUSIONOur major finding is that bloodletting at Jing-well points and MIH can reduce cerebral edema and BBB dysfunction and exert neuroprotective effects after TBI. The results suggest that the combination of BL and MIH is more effective than other treatment being used alone.

Animals ; Blood-Brain Barrier ; Bloodletting ; Brain ; pathology ; Brain Edema ; prevention & control ; Brain Injuries ; therapy ; Hypothermia, Induced ; Rats ; Rats, Sprague-Dawley

Human with bi-allelic WNT10A mutations and epithelial Wnt10a knockout mice present enlarged pulp chamber and apical displacement of the root furcation of multi-rooted teeth,known as taurodontism;thus,indicating the critical role of Wnt10a in tooth root morphogenesis.However,the endogenous mechanism by which epithelial Wnt10a regulates Hertwig's epithelial root sheath(HERS)cellular behaviors and contributes to root furcation patterning remains unclear.In this study,we found that HERS in the presumptive root furcating region failed to elongate at an appropriate horizontal level in K14-Cre;Wnt10afl/flmice from post-natal day 0.5(PN0.5)to PN4.5.EdU assays and immunofluorescent staining of cyclin D1 revealed significantly decreased proliferation activity of inner enamel epithelial(IEE)cells of HERS in K14-Cre;Wnt10afl/flmice at PN2.5 and PN3.5.Immunofluorescent staining of E-Cadherin and acetyl-α-Tubulin demonstrated that the IEE cells of HERS tended to divide perpendicularly to the horizontal plane,which impaired the horizontal extension of HERS in the presumptive root furcating region of K14-Cre;Wnt10afl/flmice.RNA-seq and immunofluorescence showed that the expressions of Jag1 and Notch2 were downregulated in IEE cells of HERS in K14-Cre;Wnt10afl/fl mice.Furthermore,after activation of Notch signaling in K14-Cre;Wnt10afl/flmolars by Notch2 adenovirus and kidney capsule grafts,the root furcation defect was partially rescued.Taken together,our study demonstrates that an epithelial Wnt10a-Notch signaling axis is crucial for modulating HERS cell proper proliferation and horizontal-oriented division during tooth root furcation morphogenesis.

OBJECTIVETo screen the high risk factors for relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT) in acute lymphocytic leukemia (ALL), acute myeloid leukemia (AML) and chronic myeloid leukemia (CML) respectively, then to compare the contribution of each risk factor to relapse and investigate the relevant mechanisms.

METHODSA retrospective study from single center involved in 262 evaluable cases of leukemia received allo-HSCT over the past 8 years, of them 69 cases with ALL, 90 AML (except APL) and 103 CML. Cox proportional hazard regression model was used for univariate and multivariate analysis to screen the high risk factors.

RESULTSThe risk factors significantly affecting relapse in ALL included: Cytogenetic risk classification, the cycles of initial induction chemotherapy; AML: Cytogenetic risk classification, minimal residual disease (MRD) level before transplant, reconstitution of WBC, and CD4(+)/CD8(+) lymphocyte ratio in the graft; CML: disease stage before transplant.

CONCLUSIONSThe relapse risk after HSCT of ALL mainly depends on the grade of malignancies, and the relapse risk of AML is closely related to the course of transplant. Chronic phase of CML favors a good prognosis after HSCT. Cytogenetic risk classification is the most relevant predictor of relapse after HSCT.

Adolescent ; Adult ; Child ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukemia ; pathology ; surgery ; Male ; Middle Aged ; Recurrence ; Retrospective Studies ; Risk Factors ; Transplantation, Homologous ; Young Adult

Objective To study the effects and mechanisms of tetramethylpyrazine(TMP)on tumor necrosis factor-α(TNF-α)-induced inflammatory injury in human coronary artery endothelial cells(HCAEC).Methods HCAEC were randomly divided into four groups:the control group(no treatment),the model group(treated with TNF-α,50 ng/mL for 24 hours),the TMP group(pre-treated with TMP,80 μg/mL for 12 hours followed by TNF-α treatment for 24 hours),and the SIRT1 inhibitor group(pre-treated with TMP and the specific SIRT1 inhibitor EX527 for 12 hours followed by TNF-α treatment for 24 hours).Cell viability was assessed using the CCK-8 method,lactate dehydrogenase(LDH)activity was measured using an LDH assay kit,reactive oxygen species(ROS)levels were observed using DCFH-DA staining,expression of pyroptosis-related proteins was detected by Western blot,and SIRT1 expression was analyzed using immunofluorescence staining.Results Compared to the control group,the model group showed decreased cell viability,increased LDH activity,ROS level and expression of pyroptosis-related proteins,and decreased SIRT1 expression(P<0.05).Compared to the model group,the TMP group exhibited increased cell viability,decreased LDH activity,ROS level and expression of pyroptosis-related proteins,and increased SIRT1 expression(P<0.05).In comparison to the TMP group,the SIRT1 inhibitor group showed decreased cell viability,increased LDH activity,ROS level and expression of pyroptosis-related proteins,and decreased SIRT1 expression(P<0.05).Conclusions TMP may attenuate TNF-α-induced inflammatory injury in HCAEC,which is associated with the inhibition of pyroptosis and activation of the SIRT1 signaling pathway.

OBJECTIVETo investigate the effects of TNF-alpha, TNF-beta and the acceptor expression about mechanical renal trauma with extraneous ADM.

METHODSThere were 104 healthy adult plain grade Wistar rat, randomly divided into four groups:8 in the group of control, 32 in the group of trauma, 32 in the group injected ADM before trauma, 32 in the group injected ADM post trauma. The experimental model of rat kidney with mechanical trauma was prepared by striking the area of rat skin reflecting by kidney with free dropping ferrous hammer in the last three groups. ADM (0.1 nmol/kg) administrated by intraperitoneal injection at 10 minutes before trauma or post trauma respectively in injected groups. All rats were executed by drawing-out all the blood in their hearts. Renal tissue was investigated to study positive expression of TNF-alpha, TNF-beta, TNFR after SABC stained.

RESULTSTNF-alpha expression:the TNF-alpha expression of trauma group was more positive than it of control group in the wound early time. The expression of group injected post trauma was less than it of trauma group at 1 h (P < 0.01). The expression of group injected before trauma was less than it of trauma group at 6 h (P < 0.05) TNF-beta expression: the TNF-beta expression of trauma group was less than it of control group at 1 h and 6 h (P < 0.05). The TNF-beta expression of group injected post trauma was more positive than it of trauma group at the same time of 1 h and 6 h (P < 0.01). TNFR expression: the TNFR expression of trauma group was less than it of control group at 6 h (P < 0.01). The TNFR expression of group injected before trauma was more positive than it of trauma group in the at the same time of 1 h and 6 h (P < 0.01).

CONCLUSIONSThe TNFR can regulate the TNF-alpha and the TNF-beta in dynamic balancing. The regulation of TNFR is main to TNF-alpha. What the TNF-beta participated in renal trauma mainly is the anti-damage process. ADM can reduce the expression of TNF-alpha. ADM increases the expression of TNF-beta and TNFR.

Adrenomedullin ; pharmacology ; Animals ; Disease Models, Animal ; Female ; Kidney ; injuries ; metabolism ; Lymphotoxin-alpha ; metabolism ; Male ; Rats ; Rats, Wistar ; Receptors, Tumor Necrosis Factor ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism