Objective To investigate the mechanism of protective effects of 17-β estradiol on the experimental model of spinal cord injury (SCI) rats.Methods First,the primary astrocytes were cultured and identified.When the third generation astrocytes were cultured,they were induced by H202 whose concentrations were established by the method of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT).The cells were randomly divided into five groups:control group; the group of treatment with 400 μmol/L H2O2 for 24 hours; the group of treatment with 20 nmol/L estrogen for 2 hours prior to exposure to 400 μmol/L H2O2 for 24 hours; the group of treatment with 20 nmol/L estrogen for 26 hours and the group of treatment with dimethyl sulfoxide for 26 hours.The proteins which were extracted from these cells after treatments with H2O2 for 24 hours were detected by Western blotting.Results The absorbances of the astrocytes of treatments with H2O2 were reduced( q' =-11.45,P =0.001 ).But exposure to estrogen prior to exposure to H2O2 provided partial restoration of the absorbances (q' =7.025,P =0.0025 ).The absorbances of the astrocytes among different groups showed significant differences( F =69.69,P =0.0025 ).The results suggested that estrogen might increase the cell viability in astrocytes.Compared with the group of treatment cells with H2O2,treatment cells with 17-β estradiol prior to H2O2 exposure down-regulated the expressions of both phosphatase and tensin homologue deleted on chromosome 10 ( PTEN ) ( F =290.003,P =0.001 ) and caspase-3 ( F =46.158,P =0.023 ).And,17-β estradiol treatment of cells increased the levels of p-Akt ( F =49.173,P =0.033 ) and Bcl-2 ( F =115.916,P =0.001 ) when compared with the group of treatment astrocytes with H2O2.Conclusion These findings suggest that the attenuation of PTEN expression mediated by estrogen is associated with an increase in phosphorylation/activation of the Akt and the Bel-2 expressions.These results suggest that the protective effects of 17-β estradiol on the experimental model of SCI rats may depend on the estrogen protection to the astrocytes which may be mediated by decreasing the PTEN expression.

OBJECTIVETo explore the correlation between F10 gene mutation and its phenotype in a Chinese pedigree affected with FX deficiency.

METHODSProthrombin time(PT), activated partial thromboplastin time(APTT), fibrinogen, FII activity(FII:C), FVII activity(FVII:C), FIX activity (FIX:C), FX activity(FX:C) were determined with a one-stage clotting assay. The FX antigen(FX:Ag) was detected with an enzyme linked immunosorbent assay(ELISA). The 8 exons, introns and 5' and 3' untranslated regions(UTR) of the F10 gene of the proband and her family members were subjected to PCR amplification and Sanger sequencing. Suspected mutation was confirmed by reverse sequencing. Polymorphisms were excluded by direct sequencing of 100 healthy individuals.

RESULTSThe PT and APTT of the proband have prolonged to 16.1 s and 49.0 s, respectively. Her FX:C and FX:Ag were reduced by 27% and 56%, and her mother's PT, APTT, FX:C and FX:Ag were 14.8 s, 37.4 s, 44%, 34%, respectively. Her grandmother's PT, APTT, FX:C and FX:Ag were 15.8 s, 42.2 s, 31%, 45%, respectively. The results of her father and other family members were all within the normal range. Genetic analysis has revealed a heterozygous G to A mutation in the proband at position 28076 in exon 8 of the F10 gene, which resulted in a p.Gly363Ser substitution. The same mutation was also found in her mother and grandmother. No mutation of the F10 gene was found in her father. Gly363Ser may result in changes in the secondary structure of the FX protein and reduction of its activity.

CONCLUSIONThe g.28076G to A(p.Gly363Ser) mutation of the F10 gene probably underlies the FX deficiency in this pedigree. The mutation was discovered for the first time in Chinese patients.

OBJECTIVETo investigate the phenotype and genotype defect characteristics of a Chinese patient with hereditary factor XI deficiency.

METHODSThe activated partial thromboplastin time (APTT), prothrombin time (PT), FXI activity (FXI:C) of the proband and his relatives were measured by a clotting method using automatic coagulation analyzer. FXI antigen (FXI:Ag) was assayed by enzyme-linked immunosorbent assay (ELISA). Fifteen exons of the F11 gene were amplified by PCR and sequenced. Pymol software was used to analyze the novel mutations.

RESULTSThe APTT of the proband was significantly prolonged (70.3 s, reference 34.5 s) with decreased FXI activity (6%, reference 50%-150%) and FXI antigen (1.9%, reference 50%-150%). The FXI activity and FXI antigen of his son was 31% and 39%, respectively. Two heterozygous F11 mutations were identified in the proband, which included a G to T substitution at nucleotide 1296 in exon 11 resulting in substitution of glycine by valine at codon 400 (p.Gly400Val) and a A to T substitution at nucleotide 1691 in exon 14 resulting in substitution of arginine (AGA) by a termination codon (TGA) at codon 532 (p.Arg532Ter). Analysis using Pymol indicated that the number of hydrogen bonds has changed, which led to a transformation of the structure of the FXI protein. The son of the proband was found to be heterozygous for the c.1296G to T (p.Gly400Val) mutation. NM_13142 c.1691A to T (p.Arg532Ter) is a novel mutation based on HGMD professional 2016.4. Based on 2015 Guidelines of ACMG, it is PVS1 (very strong pathogenicity).

CONCLUSIONThe compound heterozygous mutations of F11 NM_13142 c.1296G to T (p.Gly400Val) and F11 NM_13142 c.1691A to T(p.Arg532Ter) probably underlies the FXI deficiency in the proband.

Objective:To analyze the clinical manifestations and duration of feeding intolerance (FI) in very low birth weight infants (VLBWI), explore the influencing factors of FI, so as to provide scientific basis for shortening the FI duration of VLBWI.Methods:From January 2018 to December 2020, 441 VLBWIs admitted to the Neonatal Intensive Care Unit of the Shandong Provincial Hospital Affiliated to Shandong First Medical University were selected as the research object by the convenient samplingto describe the clinical characteristics and duration of FI in children. Multivariate Logistic regression was used to analyze the influencing factors of FI duration of VLBWI. Totally 354 VLBWIs were included, excluding 41 children transferred from other hospitals 24 hours after birth, 10 children died before full gastrointestinal feeding, 4 children with stageⅡ or above neonatal necrotizing enterocolitis, 1 child with congenital gastrointestinal malformation, and 31 cases with incomplete data.Results:The incidence of FI in 354 VLBWI was 71.5% (253/354), and the average duration was 9 days. Multivariate Logistic regression analysis showed that the long time of invasive positive pressure ventilation [ OR=1.081, 95% CI (1.034, 1.130) ], formula feeding [ OR=1.751, 95% CI (1.009, 3.040) ] and hypertension during pregnancy [ OR=1.876, 95% CI (1.073, 3.279) ] were the risk factors for FI duration ( P<0.05), andbirth weight [ OR=0.355, 95% CI (0.185, 0.683) ]was the influencing factor of FI duration ( P=0.002) . Conclusions:The incidence of FI in VLBWI is high and the duration is long. Small gestational age and low weight premature infants are the high-risk groups of FI. In order to shorten the FI duration of VLBWI, medical and nursing staff should manage the blood pressure of pregnant women during pregnancy, strengthen the nursing of children during mechanical ventilation, withdraw the machine as soon as possible, and promote breastfeeding.

Objective To study the medication law of postoperative treatment of colorectal cancer by national TCM doctor Professor Piao Bingkui.Methods Professor Piao Bingkui's electronic medical records and paper medical records of colorectal cancer postoperative patients at Guang'anmen Hospital,China Academy of Chinese Medical Sciences and Beijing Yiqingtang Chinese Medicine Clinic were collected and organized from their outpatients from January 1st,2002 to February 28th,2022.R4.2.1 was used to study the prescriptions,including high-frequency drugs,drug types,properties of Chinese materia medica,yin yang and five elements and dosage of drugs,as well as the law of multi-drug association in postoperative patients with colorectal cancer.Results Totally 642 colorectal postoperative cancer patients were included,involving 2 226 prescriptions,180 kinds of Chinese materia medica,and a total frequency of 39 988 times.The high-frequency drugs were Astragali Radix,fried Aurantii Fructus with wheat bran,Dioscoreae Rhizoma,etc.The main drugs in terms of efficacy were tonics for tonifying deficiency,disinfectants,etc.;the main properties were warm and neutral,the main tastes were sweet,pungent and bitter,and the main meridians were spleen meridians and stomach meridians.Ascending medicines were often used,and the drug were basically non-toxic.The frequency of using yang tonifying medicine was high,and the five elements were commonly used as local medicines;the dosage was mostly 10,15 and 20 g.The complex network analysis and clustering analysis of the association between multiple drugs found that Professor Piao Bingkui's basic prescription for treating colorectal cancer included Astragali Radix,fried Aurantii Fructus with wheat bran,Dioscoreae Rhizoma,salt Alpiniae Oxyphyllae Fructus,Citri Reticulatae Pericarpium,Smilacis Glabrae Rhizoma,Pseudostellariae Radix,and fried Atractylodis Macrocephalae Rhizoma with wheat bran.Conclusion In the treatment of colorectal cancer,Professor Piao Bingkui focuses on reinforcing the healthy qi,nourishing spleen and stomach function,combined with detoxication method and clearing heat,expelling phlegm and dampness,guiding stagnation and dispelling stasis methods,making syndrome differentiation as well as tonifying and benefiting qi,regulating qi movement,so as to realize the"treating middle-energizer as balance"and achieve mild level when treating colorectal cancer.

Objective To investigate the effect of MMP-8 on cornea. Methods Fifteen C57BL/6J healthy mice were selected. The right eyes corneal stroma was injected by 10μL MMP-8 as the experimental group and the left eyes were injected by same amount of normal saline as the control group. At 0,4,8 h, the two-photon microscope second harmonic generation imaging technology was used to scan mice corneal stroma layer by layer in vivo. The obtained images were performed the 3D reconstruction by Imaris software and the signal intensity of the images were calculated. At 4,8 h, the corneal opacity degree was evaluated under slit lamp. At 8 h,mice were killed and corneas were collected to determine the hydroxyproline concentration. Results The cornea stromal fiber signal strengthes at 0 h in the experimental group and control group were (89.7±11.2) and (85.3±7.0),which at 4 h were (14.5±3.4) and (46.6±14. 0) respectively,which at 8 h were (11.0±4.6) and (34.6±12.5) respectively. The cornea stromal signal strength at 4,8 h in the experiemental group was significantly decreased compared with that in the control group (P<0.05) ;the cornea at 4 ,8 h in the experimental group was significantly turbid than that in the control group (P<0.05);the cornea hydroxyproline concentrations detected at 8h in the experiemental group and control group were (0.433±0. 090) μg/mg and (0. 590±0. 133) μg/mg respectively,the experimental group was significantly lower than the control group (F=7. 193,P=0. 014). Conclusion MMP-8 has obvious degradation and destroy effect on mice corneal stroma collagen,which leads to the decrease of corneal opacity.