Objective To investigate the corelation between platelet activating factor acetylhydrolase (PAF-AH) genetic polymorphism and ischemic stroke. Methods The plasma PAF-AH genotype was determined in 205 patients with iachemic stroke and 114 normal subjects by the polymerase chain reaction. The levels of plasma platelet activating factor (PAF), platelet α-granule membrane glycoprotein-140(GMP-140), β-thromboglobulin (β-TG) and the levels of platelet factor 4 (PF4) were analyzed. Results The prevalence of the mutation genotype and plasma PAF, GMP-140, β-TG and PF4 in the patients with isehemic stroke [42.44%,(91.08 ± 39.10) ng/L, (36.46 ± 13.10) μg/L, (41.75 ± 11.18) μg/L, (29.05 ± 9.16) g/L, respectively] were significantly higher than those in the controls[21.05%,(64.30 ± 18.81) ng/L, (18.27 ± 7.68) μg/L, (30.94 ± 8.47) μg/L, (18.75 ± 6.06) μg/L](P< 0.01). The levels of plasma PAF, GMP-140 were significantly higher in mutation genotype patients than those in the normal genotype patients (P < 0.01). Conclusions The activation function of platelet in the acute phase of patients with ischemic stroke increases, and it is associated with genetic polymorphism of PAF-AH. The PAF-AH gene mutation may be a novel genetic marker for high risk of ischemic stroke.

Objective To investigate the effect of electroacupuncture at points Xiaohai and Xiajuxu on the expressions of tumour necrosis factor-α (TNF-α) and choline acetyltransferase (ChAT) in serum and nicotinic acetylcholine receptorα 7 (α 7 nAchR) in duodenal tissues in a rat model of duodenal ulcer (DU) and preliminarily explore the relative specificity of He-Sea point in “treating visceral diseases with He-Sea point”.Methods Forty healthy SD rats were randomized into blank (A), model (B), Xiaohai (C) and Xiajuxu (D) groups, 10 rats each. A rat model of DU was made by subcutaneous injection of 10% cysteamine hydrochloride at the right buttock. After successful model making, group C was given electroacupuncture at point Xiaohai and group D, at point Xiajuxu. Duodenal tissue ulcer was macroscopically observed and scored in every group of rats. Rat serum expression of TNF-α was determined by double antibody sandwich enzyme-linked immunosorbent assay (ELISA); rat serum expression of ChAT, by ultraviolet spectrophotometry & colorimetry; rat duodenal expression ofα7 nAchR, by Western blot.Results After model making, the duodenal ulcer score was significantly higher in groups B, C and D than in group A (allP<0.01) and significantly lower in groups C and D than in group B (bothP<0.01) and in group D than in group C (P<0.01). TNF-α expression was significantly higher in group B than in group A (P<0.01) and significantly lower in groups C and D than in group B (bothP<0.01) and in group D than in group C (P<0.01). ChAT expression tended to increase in group C compared with group B but there was no statistically significant difference (P>0.05) and was significantly higher in group D than in group B (P<0.01) or C (P<0.05).α7 nAchR expression was significantly higher in groups C and D than in group B (bothP<0.01). There was a positive correlation between ChAT andα7 nAchR expressions in every group (r=0.444,P=0.007).Conclusions Electroacupuncture at both points Xiaohai and Xiajuxu can reduce the duodenal ulcer score and serum TNF-α expression and increase serum ChAT and duodenalα7 nAchR expressions in DU rats. The results show that the therapeutic effect of electroacupuncture on duodenal ulcer may be produced by regulating TNF-α. Its mechanism may be activating cholinergic anti-inflammatory pathway to produce an anti-inflammatory effect. The effect being better in group D than in group C suggests that Xiajuxu has the relative specificity.

Cell culture plays an important role in virology. It provides a platform for the detection and isolation of viruses as well as for the biochemistry and molecular biology based studies of viruses. In the present work, a new system that could permits multiple (different) cell lines to be simultaneously cultured in one dish was developed. In the system, each cell line was cultured in an isolated zone in the same dish or well and the system is therefore called an isolated co-culture system. The usefulness of this novel approach for virus isolation was demonstrated using a model system based on adenovirus.

The cystine knot (CK) motif comprises an internal ring formed by two disulfide bonds and their connecting backbone segments which is threaded by a third disulfide bond .It is present in peptides and proteins of a variety of species ,in-cluding fungi ,plants ,marine molluscs ,insects and spiders .CK polypeptide is one of the ideal model molecules for drug design and molecular engineering research because of its stable structure and variety of bioactivities .Here we summarized the main structural features of both inhibitor cystine knot (ICK) peptide and cyclic cystine knot (CCK) peptide ,including primary se-quence ,topology ,permutation ,synthesis and folding characteristics ,as well as its applications on drug design and molecular engineering .

Objective To evaluate the effect of diammonium glycyrrhizinate extracted from the Chinese traditional medicine licorice root on the growth of human hair follicles cultured in vitro,and to detect the expression of wnt/β-catenin signaling pathway-related molecules.Methods Isolated hair follicles were cultured with diammonium glycyrrhizinate at different concentrations of 0.1,0.01,0.001 and 0.000 1 μmol/L for 10 days,and the hair follicles cultured in Williams' E medium without diammonium glycyrrhizinate served as a control group.The length of hair follicles was measured under a microscope every day,the morphologic changes of hair follicles were observed,and photos were taken.Immunofluorescence assay was performed to assess the proliferation of hair matrix cells,as well as to determine the expression of β-catenin,glycogen synthase kinase 3β (GSK3β),phosphorylated GSK3β (p-GSK3β) and lymphoid enhancer factor-1 (Lef1) in the Wnt/β-catenin signaling pathway.Statistical analysis was carried out by repeated-measures analysis of variance and one-way analysis of variance.Results As repeated-measures analysis of variance showed,only 0.01 μmol/L diammonium glycyrrhetate showed significantly promotive effect on the growth of hair follicles compared with the medium alone (P ＜ 0.05),and there were no significant differences in the length of hair follicles between the other concentration groups and the control group.Compared with the control group,the transition to the catagen phase of human hair cycle was delayed in the 0.01-μmol/L diammonium glycyrrhetate group,while it did not change in the other diammonium glycyrrhetate groups and control group.Immunofluorescence assay showed that the number of ki67-positive hair matrix cells was obviously increased in the 0.1-,0.01-,0.001-μmol/L diammonium glycyrrhizinate groups compared with the control group,while there was no difference between the 0.000 1-μmol/L diammonium glycyrrhizinate group and the control group.One-way analysis of variance revealed that the expression of β-catenin,p-GSK3β and Lef1 significantly differed among all the groups (F =12.604,16.65,15.266 respectively,P ＜ 0.05),while no significant difference in the expression of GSK3β was found among these groups (F =1.472,P ＞ 0.05).Least significant difference (LSD)-t test revealed that the expre-ssion of β-catenin,p-GSK3β and Lef1 in the hair matrix cells was significantly higher in the 0.1-,0.01-,0.001-μmol/L diammonium glycyrrhizinate groups than in the control group (all P ＜ 0.05),but there was no significant difference between the 0.000 1-μmol/L diammonium glycyrrhizinate group and the control group (P ＞ 0.05).Conclusion Diammonium glycyrrhetate at the concentration of 0.01 μmol/L shows markedly promotive effect on the in vitro growth of hair follicles,and can increase the proliferative activity of hair matrix cells and delay the transition to the catagen phase,which may be associated with the activation of Wnt/β-catenin signaling pathway.

Nonalcoholic fatty liver disease (NAFLD) is a major cause of chronic liver diseases worldwide and covers a series of pathological manifestations from hepatic steatosis to inflammation, fibrosis, and even liver cirrhosis. NAFLD is associated with a wide range of extrahepatic diseases, including metabolic syndrome, cardiovascular disease, chronic kidney disease, endocrine diseases, obstructive sleep apnea-hypopnea syndrome, psoriasis, and skeleton-muscle diseases. The major causes of death in patients with NAFLD include cardiovascular disease, malignancies, and liver-related complications, suggesting that extrahepatic diseases associated with NAFLD should be taken seriously by clinicians. This article reviews the research advances in extrahepatic diseases associated with NAFLD, so as to provide ideas for clinical assessment and treatment.

Objective To investigate the clinical characteristics and the experience of multi-disciplinary team (MDT) on recurrence of primary hyperoxaluria (PH) type I after renal transplantation. Methods One case presenting with unexplained rapid decline of renal allograft function after allogeneic renal transplantation was discussed by MDT. The role of MDT in diagnosing rare hereditary diseases and improving the long-term survival of renal transplant recipients was summarized. Results After MDT consultation, the patient was diagnosed with recurrence of PH type I. Routine immunosuppressive regimen was initiated after the exclusion of rejection. The patient was instructed to drink a large quantity of water, and given with high-quality protein and low-phosphorus diet, vitamin B6, calcium and other conservative therapies to actively prevent and treat postoperative complications. The deterioration of renal graft function was delayed. Nevertheless, regular hemodialysis was resumed at 5 months after renal transplantation until the submission date of this manuscript. Conclusions Recurrence of PH type I after renal transplantation is relatively rare. The main clinical manifestations are recurrent kidney stones and decreased renal function with multiple complications and poor prognosis. The condition of the patient is consulted by MDT for confirming the diagnosis, determining the optimal treatment scheme, delaying the progression and improving the clinical prognosis.

Objective:To summarize the clinical characteristics of central nerve system (CNS) infection and grasp the necessity and possibility of early diagnosis and precise intervention of CNS infection after renal transplantation.Methods:This retrospective study enrolled consecutive recipients of renal transplantation with CNS infection after transplant between January 2000 and December 2020. Correlative factors for CNS infection after renal transplant were determined by comparing the clinical data between recipients with and without CNS infection. After screening 3, 199 consecutive renal transplant recipients, 12 patients with CNS infection post-transplant were identified and recruited. The median age-of-onset was 48.5 (23-65) years. And the median time to disease onset after transplant was 50.5(1-204) months. The most common symptoms of CNS infection after renal transplant included fever (75.00%), consciousness disorder (58.33%), headache (58.33%) and neck rigidity (41.67%).Results:Hepatitis B virus carrier and pulmonary infection were correlated with CNS infection after transplantation ( P<0.05). Nine patients failed to identify the pathogen and only received empirical anti-infective regimen. The outcomes were curing ( n=3) and death ( n=6). Metagenomic sequencing was performed for identifying the pathogen in three recipients and actively adjusting the anti-infective regimen. As a result, 2 were cured and 1 died. The overall mortality was 58.33%. The median time to death or curing from disease onset were 20(2-19) and 25(16-35) days respectively in surviving and non-surviving recipients. Conclusions:The progress of CNS infection after transplantation is rapid with a high mortality. HBV carrier and pulmonary infection are possible risk factors of CNS infection after renal transplantation. Early pathogenic identification and precise etiological intervention are vital for better clinical outcomes.

Background and objective Non-small cell lung cancer (NSCLC) presents the highest morbidity and mortality among malignant tumors worldwide. hTe overall effective rate of epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) is 30% to 40%, and PFS (progression-free sruvival) is 12 months. However, EGFR-TKI resistance is typical in clinical observations, and this phenomenon signiifcantly affects tumor suppression. To overcome this resistance, a new prognostic factor associated with lung cancer drug resistance should be discovered. This study investigated the rela-tionship between the inhibitor of differentiation 1 (ID1) and non-small cell lung cancer EGFR-TKI resistancein vivo andin vitro to determine any statistical signiifcance and discuss the underlying mechanism.Methods Western blot and qRT-PCR were used to quantify the expression of ID1 in lung cancer. IHC was used to detect the expression of ID1 in pathological tissues (lung cancer tissues and adjacent tissues). MTT was used to detect cell proliferation, in which the cells were treated with geiftinib atfer being transfected by ID1 slow virus vector. Lung cancer cells were inoculated in nude mice until the tu-mor diameter grew to certain measurement. Geiftinib treatment was started, and the tumor volume was estimated.Results ID1 was highly expressed in NSCLC (P<0.05). Both ID1 expression and drug resistance of EGFR-TKI in NSCLC were positively correlated (P<0.05). hTe treatment group with geiftinib showed obviously less expression than the control group. Conclusion ID1 is highly expressed in NSCLC. ID1 expression was positively related to drug resistance of EGFR-TKI in NSCLC. Geiftinib can be used to effectively treat NSCLC, and the mechanism may be associated with an increased level of STAT3 phosphorylation.

Objective To explore prenatal ultrasonic characteristics of fetal Currarino syndrome(CS)and methods for prenatal diagnosis of CS.Methods Two fetuses with CS confirmed by genetic examination were retrospectively analyzed,while 6 CS fetuses with complete prenatal ultrasonic data in literature were reviewed.Prenatal ultrasonic characteristics of CS fetuses and the method for prenatal diagnosis of CS were discussed.Results Among 8 CS fetuses diagnosed with prenatal ultrasound,4 were female singletons with a clear family history of CS,and MNX1 gene mutation was found in 1 fetus.The other 4 fetuses were 2 pairs of male monochorionic twins,all with MNX1 gene mutation.Among 8 CS fetuses,complete triad(sacral agenesis abnormalities,anorectal malformation and presacral mass)were displayed only in 2 fetuses,while all 8 had sacral agenesis abnormalities and 6(6/8,75.00％)were detected with prenatal ultrasound,6 had low location of conus medullaris and 2(2/6,33.33％)detected with prenatal ultrasound.Conclusion Prenatal ultrasound was the first choice for non-invasive diagnosis of fetal CS.When one of sacral agenesis abnormalities,anorectal malformation and presacral mass was found with prenatal ultrasound,the possibility of CS should be considered,and fetal MRI,genetic examination and prenatal genetic counselling should be recommended if necessary.