Epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) is one of the most important targeted drugs for lung cancer patients carrying EGFR sensitive mutations.However,almost all patients that are effective to this treatment will eventually develop secondary resistance to EGFR-TKI.The most accepted mechanisms of resistance mainly include T790M mutation,MET amplification,PIK3CA mutation,down-regulation of PTEN expression and activation of Fas-transcription factor-κB.Recent years,many new drugs are developed to overcome this resistance.Although most of drugs are in the stages of cell or animal experiment,some new drugs get positive clinical results.

Objective To study the relationship between the complete blood count (CBC) and coal‐buring caused fluorosis of fe‐male rats .Methods Female SD rats were randomly divided into three groups :control group ,medium‐fluorine group and high‐fluo‐rine group .Rats in each exposed group were fed with fodder containing different proportions of corn dried by burning coal from flu‐orosis endemic areas to establish coal‐burning fluorosis model (fluoride of fodder were 47 .8 mg/kg and 96 mg/kg) .The corn of control group′s fodder was collected from non endemic areas (fluoride was 5 .2 mg/kg) .At 60 days ,120 days and 180 days ,the tail vein bloods were analyzed with automated analyzer .Results Compared with medium‐fluorine group ,the WBC of high‐fluorine group decreased at 60 d and 120 d(P<0 .05) .Compared with control group ,the RBC of fluoride treated groups decreased at each time point (P<0 .05) ,especially at 120 d .At 60 d and 120 d ,the Hb ,HCT and MCV decreased(P< 0 .05) .At 180 d ,only the MCV of high‐fluorine group increased obviously(P<0 .05) .At 120 d ,the Hb ,HCT of fluoride treated groups were less than those at 60 d and 180 d (P<0 .05) .The MCV of high‐fluorine group was same as above(P<0 .05) .At 180 d ,the MCV of medium‐fluo‐rine group decreased less than that at 60 d (P< 0 .05) .The MCH of fluoride treated groups increased at each time point (P<0 .05) .At 120 d ,the MCH of medium‐fluorine group increased more than its at 60 d and 180 d (P<0 .05) .At 180 d ,the MCH of higher‐fluorine group increased than those at 60 d (P<0 .05) .Conclusion Fluorosis has varied influence on blood cell of SD rats , especially on red blood cell system .In the early and mid stages ,the coal‐buring caused fluorosis showed the small RBC high pigment anemia .In the late stage ,the coal‐buring caused fluorosis showed the big RBC high pigment anemia .

Objective To observe the influence of coal-burning type of fluorosis on hypothalamic-pituitaryovary axis function and to explore possible mechanism in female rats.Methods Sixty SD rats were divided into two groups according to body weight with the method of random number table:control group and fluorosis group,30 rats in each group.Fluorosis group was feed with corn powder baked by high fluorine coal from Zhijin area.Changes of female rats' teeth during fluorine exposure were observed.After feeding for 180 days,24 h urine was collected in estrus and fluorine level was tested using fluoride ion-selective electrode; rats were executed and bone fluorine level was tested with high-temperature ashing-fluorine ion-selective electrode.Femoral artery blood was collected and serum was separated to test the contents of follicle stimulating hormone (FSH),luteinizing hormone (LH),testosterone (T),estradiol (E2) and progesterone (P) with electrochemiluminescence radioimmunoassay and gonadotropin-releasing hormone (GnRH),inhibin (INH) with enzyme-linked immunosorbent assay (ELISA),respectively.Organs,including hypothalamus,pituitary gland and ovary were weighted,and organ coefficients were calculated.Pathological morphology of hypothalamus,pituitary gland and ovary was observed after staining and ultrastructure of ovary was examined by electron microscopy.Results Coal-burning induced fluorine poisoning rat model was established successfully.There were no significant differences statistically in organ coefficients between fluorosis groups (0.032 ± 0.004,0.014 ± 0.008,0.037 ± 0.009) and controls (0.035 ± 0.005,0.012 ± 0.006,0.035 ± 0.004,t =0.46,0.87,0.64,all P ＞ 0.05).Rats serum GnRH,FSH,LH and T levels [(21.654 ± 4.765),(29.580 ± 5.221),(53.988 ± 6.506),(23.962 ± 2.255)μg/L] of fluorosis groups were all higher than those of controls [(10.384 ±2.250),(19.217 ± 4.743),(30.314 ± 4.443),(7.883 ± 1.973)μg/L,t =6.762,4.646,9.503,16.971,all P ＜ 0.05].But the level of P,INH [(12.635 ± 3.841),(18.926 ± 3.465)μg/L] were all lower than those of controls [(21.045 ±4.768),(48.076 ± 3.525)μg/L,t =4.344,18.649,all P ＜ 0.05].Serum E2 levels of control group and fluorosis group were (35.375 ± 10.662) and (27.500 ± 12.783)μg/L,respectively.The difference between groups was not statistically significant (t =1.821,P ＞ 0.05).No pathological changes were observed in the two groups of female hypothalamus,pituitary tissue by light microscopy and electron microscopy.Under light microscope,in the control group of normal ovarian tissue,more corpus luteum and different developmental stages of follicles were seen,granulosa cells were neatly arranged in a monolayer or multilayer.In fluorosis group,severe edema of ovarian interstitial cells and follicle degeneration increased.Cell structure and cell contours were blurred and unclear with occasional mature follicles.Under transmission electron microscope,in control group,normal ovarian granulosa cell ultrastructure was observed,nuclei were round,nuclear chromatin was uniform distributed,cytoplasm was rich in mitochondria and endoplasmic reticulum,and normal morphology.In fluorosis group,granulosa cells and interstitial cells showed apoptotic characters,such as nucleoli disappearing,mitochondrial swelling and chromatin aggregating at the nuclear membrane.Conclusions Fluorosis can induce ovarian tissue apoptosis,severely damage the micro environment.Reduction of P and INH affects ovarian,maturation and ovulation and leads to secretion of GnRH,FSH and LH.Fluorosis caused by coal-burning may induce the injury of ovary and cause abnormal secretion of hypothalamic-pituitary-ovary axis.Fluorosis has affected parts of female axis which may not be in the hypothalamus,pituitary,but causes ovarian tissue damage.

Objective To observe the impact of coal-burning-borne fluorosis on the opening day of vagina,and estrogen cycle and endocrinology of female rat.Methods Ninety weaned two weeks and healthy female SD rats,weighing 60-80 g were selected,and divided into three groups by body weight using random number table:control group,moderate fluoride exposed group,and higher fluoride exposed group,thirty rates in each group.Rats in fluoride exposed groups were fed with baked corn collected from coal-burning-borne fluorosis areas containing different levels of fluorine(fluorine contents were 47.8 and 96.0 mg/kg,respectively) to establish coal-burning-borne fluorosis models.The corn fed to control group rats were collected from non-endemic areas (fluoride content was 5.2 mg/kg).After exposed to fluoride for 60,120 and 180 days,the rats were put to death during estrus in three batches (10 rats in every batche).The situation of female vaginal opening was observed and recorded every morning at 8:00 at the beginning of the experiment.Ten female rats in each group were randomly selected to observe vaginal exfoliated cells for 10 days continuously every morning,and HE staining was selected to discriminate estrogen cycle.Femoral artery blood was collected to observe the impact of estradiol(E2) in estrus.Results The vaginal opening day age of the longer fluoride exposed group [(49.0 ± 4.5)d] was longer than that of the control group[(45.4 ± 2.3)d] and the moderate fluoride exposed group[(46.7 ± 1.9)d,all P ＜ 0.05].Estrogen cycle of the longer fluoride exposed group[(5.8 ± 0.8)d] after fluoride exposure for 120 d was higher than that of the control group [(4.4 + 0.9)d] and the moderate fluoride exposed group [(4.9 ± 0.7)d,all P ＜ 0.05].Estrogen cycle increased with increasing dosage of fluoride after fluoride exposure for 180 d in three groups [control group:(4.3 ± 0.7)d,moderate fluoride exposed group:(5.4 ± 1.1)d,higher fluoride exposed group:(6.7 + 0.9)d,all P ＜ 0.05].Estrogen cycle in higher fluoride exposed group at 180 d[(6.7 ± 0.9)d] was higher than that of 60 d and 120 d[(5.2 ± 1.1),(5.8 ± 0.8)d,all P ＜ 0.05].At 60 d after exposed to fluoride,E2 levels of fluoride exposed groups were increased[control group:(30.125 ± 94.17)ng/L,moderate fluoride exposed group:(438.75 ± 71.80) ng/L],higher fluoride exposed group (456.25 ± 57.80)ng/L with increasing dosage of fluoride (all P ＜ 0.05).E2 level in rats exposed to fluoride for 180 d [(275.00 ± 127.83)ng/L] was lower than that of 60 d and 120 d [(456.25 ± 57.80),(392.50 ± 102.50)ng/L,all P ＜ 0.05] in higher fluoride exposed groups.Conclusions Coal-burning-borne fluorosis has a significant effect on vaginal opening day age,estrogen cycle and endocrinology of female rats.The higher the fluoride dose,and the longer the exposure time,the more severe the impact.

Objective To apply color Doppler flow imaging (CDFI) and transcranial Doppler (TCD) to the follow-up observation of the changes of cervical vessel,intracranial hemodynamics and cerebrovascular reserve capacity (CVR) of the patients after carotid artery stent implantation.Methods Totally 96 patients with carotid artery stent implantation underwent CDFI and TCD examinations,and the changes of hemodynamics were compared before and 1 month,6 months,1 a and 2 a after implantation.Results The values of peak systolic velocity (PSV) and resistance index (RI) at the areas of carotid stenosis were lower significantly than those before implantation,while the values of PSV,pulsatility index (PI) and CVR of the middle cerebral artery were obviously higher than those before implantation (P＜0.05).There were no significant differences between the cervical and intracranial hemodynamics indexes 1 month,6 months,1 a and 2 a after treatment.Two-year follow-up found 4 cases of restenoses after implantation,and the rate for restenosis was 4.2％.Conclusion Carotid artery stent implantation improves significantly cervical and intracranial blood supply as well as CVR of the carotid stenosis patient,and CDFI combined with TCD can be used for the accurate evaluation of the efficacy and postoperative follow-up of carotid artery stent implantation.

The authors of this paper confirm the goal, factors and principles in arrangement of clinical practice, design a transplacement algorithm to solve the model, and after estimating the capacity to interns of affiliated hospital, make the schedule by that algorithm so as to relieve the relative short supply of good clinical education resourses.

OBJECTIVE To explore the expression of Maspin in invasive fungal rhinosinusitis (IFRS) and the value of Maspin in the diagnosis of IFRS. METHODS Forty two cases of fungal rhinosinusitis (FRS) were set as the experimental group, which included 12 cases of IFRS and 30 cases of noninvasive fungal rhino-sinusitis (NIFRS). At the same time, 30 cases of chronic rhino-sinusitis were set as control group. Immunohistochemistry (IHC) was used to detect the expression of Maspin. RESULTS Compared with the control group, the expression of Maspin in FRS group decreased statistically (t=-3.367, P<0.05). The IFRS group, compared with other two groups, had the lowest expression of Maspin (t=-3.390, P<0.05; t=-4.143, P<0.05). By using Maspin score of 5.70 as the cut-off point, the sensitivity and specificity for the diagnosis of IFRS was 91.7% and 88.3% respectively. CONCLUSION The expression of Maspin is very low in IFRS group. Down-regulation of Maspin expression may be a potential indicator for diagnosis of IFRS.

Objective:To summarize the characteristics of genetic variation and prenatal diagnosis in pedigrees with X-linked adrenoleukodystrophy (X-ALD) and elucidate the value of prenatal diagnosis in preventing the birth of children with X-ALD.Methods:Twenty pedigrees, clinically diagnosed with X-ALD in Peking University First Hospital from November 2012 and March 2019, were included in this retrospective study. Genomic DNA was extracted from peripheral blood and amniotic fluid or chorionic villi samples of probands and their families for detecting variants in ATP-binding cassette subfamily D member 1 ( ABCD1) gene using polymerase chain reaction (PCR)-Sanger sequencing. Linkage analysis was also performed on five microsatellite markers near ABCD1 gene to exclude maternal contamination. Characteristics of ABCD1 gene variants and prenatal diagnosis of X-ALD pedigrees were summarized by descriptive statistics. Results:Twenty ABCD1 gene variants were identified in the 20 pedigrees. The variants in three probands that were not detected by next-generation sequencing were identified by PCR-Sanger sequencing. Among the mothers of the 20 probands, 17 carried ABCD1 variants and three did not. We performed 24 prenatal diagnoses on 20 pregnancies (24 fetuses) and identified eight fetuses with variants who were finally terminated. The 16 cases without variants were born alive. The validation results obtained after termination or delivery were consistent with those performed prenatally. Conclusions:No hotspot variants in ABCD1 gene are detected in these X-ALD patients and most variants are maternally inherited. PCR-Sanger sequencing is an effective method for detecting ABCD1 variants. Prenatal diagnosis for mothers who had a body with X-ALD could prevent another one from birth.

Objective To summarize the nursing experience of delayed closure of chest with acute renal injury after switch operation and underwent peritoneal dialysis in neonates and to improve the therapeutic effect. Methods To summarize the curative effects and perioperative nursing experience of one case of the complete transposition of great arteries with intact interventricular septum neonate who underwent delayed closure of chest with acute renal injury and peritoneal dialysis after Switch operation under general anesthesia and extracorporeal circulation in November 2017 in our department. Results The child was postponed to close the chest after surgery. Low cardiac output syndrome and acute renal function injury occurred 1 hour after operation. Through monitoring hemodynamic indexes during ICU, the child recovered after timely treatment of low cardiac output syndrome, maintaining stabilization of circulation, diuresis, peritoneal dialysis, keeping water, electrolyte and acid-base balance, nursing care for delayed closure of chest and other related treatment. Postoperative assisted mechanical ventilation time was 168 hours, postoperative ICU hospitalization time was 12 days, and postoperative total hospitalization time was 19 days. Conclusion The infants who have complete transposition of the great arteries and the intact interventricular septum after Switch operation have many complications and rapid changes in the state of illness. Rigorous and meticulous nursing plays a key role in reducing the postoperative complications and improving the achievement ratio of the operation.

OBJECTIVETo analyze the characteristics of germline mutations of adenomatous polyposis coli (APC) gene in pedigrees affected with familial adenomatous polyposis (FAP).

METHODSGenomic DNA was extracted from peripheral blood samples from members of the 13 FAP pedigrees. Multiplex ligation-dependent probe amplification (MLPA) was used to detect large fragment deletions of the APC gene. Subsequently, potential mutation was screened from all exons of the APC gene with PCR amplification and direct sequencing.

RESULTSGermline mutations have been identified in 5 FAP pedigrees, which included c.3184_3187delCAAA, c.5432C>T, c.3925_3928delAAAA and c.3925_3929del AAAAG(in two pedigrees). Small deletional mutations were found primarily in the area of AAAAG tandem repeat sequences.

CONCLUSIONC.3925_3929 located in AAAAG tandem repeats is probably the hot spot for APC gene mutations, which are mostly deletional mutations, especially the 5 bp base deletion at codon 1309.

Adenomatous Polyposis Coli ; genetics ; Adenomatous Polyposis Coli Protein ; genetics ; Adult ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Female ; Humans ; Male ; Molecular Sequence Data ; Pedigree ; Sequence Deletion