Objective The study aimed to explore the relationship between AIF related pathway and the inju-ring of cultured SGNs (spiral ganglion neurons)by glutamate toxicity,and to find AIF expression and distribution changes in SGNs.Methods SGNs of 40 newborn rats within 3 day were obtained and cultured in vitro.Cultured cells were divided into four groups:the normal control group,10 mM,20 mM and 40 mM glutamate injured group, separately.After 48 h hours culturing,optical microscopy,immune fluorescence staining and real-time fluores-cence quantitative PCR were used to observe the morphology,AIF distribution,and AIF,calpain,Caspase3 expres-sion changes in SGNs in vitro.TUNEL was used to verify the cell apoptosis.ResuIts Noticeable morphological chan-ges and cell apoptosis were occurred in 20 mM glutamate group,with AIF nuclear translocation.AIF gene expression was significantly higher than normal after glutamate administration (P<0.05);moreover,calpain gene expression increased(P<0.05);but caspase3 expression was not statistically significantly increased in all glutamate treated groups (P>0.05). ConcIusion In the process of cultured SGNs injured by glutamate,AIF participated in the cell apoptosis.Noticeable cell apoptosis were occurred in 20 mM glutamate group with AIF nuclear translocation.Calpain up-expression also contributed to excitatory neurotransmitter injury on SGNs,but Caspase 3 had no obvious effects.

Objectiye To investgate the effects of isoflurane preconditioning on expression of Toll-like receptor 4 (TLR4) and myeloid differentiation factor 88 (MyD88) during focal cerebral ischemia-reperfusion (IR) in rats. Methods Thirty male SD rats weighing 250-300 g were randomly divided into 3 groups ( n = 10 each):sham operation group (group S);focal cerebral IR group and isoflurane preconditioning group (group IP). The animals were anesthetized with intraperitoneal pentobarbital 40 mg/kg. In group IR and IP a nylon thread with rounded tip was inserted into right internal jugular vein and threaded cranially until resistance was met. Mid-cerebral artery was occluded (MCAO) for 2 h followed by 24 h reperfusion. In group IP the animals inhaled 2% isoflurane98 % O2 for 1 h once a day for 5 consecutive days at 24 h before MCAO. Neurologic function was assessed and scored and cerebral infarct volume was measured at 24 h of reperfusion. The animals were sacrificed at 24, 48 and 72 h of reperfusion respectively. The right ischemic frontal lobes were removed for determination of TLR4, MyD88and NF-κB expression by Western blot analysis. Results MCAO significantly worsened neurologic function. The neurologic function deficit scores were significantly increased and the TLR4, MyD88 and NF-κB expression were significantly up-regulated in group IR as compared with group S (P ＜ 0.05). Isoflurane preconditioning significantly decreased cerebral infarct volumes and neurologic function deficit scores and down-regulated the expression of TLR4, MyD88 and NF-κB in group IP as compared with group IR ( P ＜ 0.05). Conclusion Isoflurane preconditioning can reduce inflammatory response and focal cerebral IR injury by down-regulating the expression of TLR4and Myd88.

[ ABSTRACT ] AIM: To investigate the expression and clinical significance of bone morphogenetic protein 3 (BMP3) in hilar cholangiocarcinoma tissues.METHODS: Thirty cases of hilar cholangiocarcinoma specimens were col-lected.The expression of BMP3 at mRNA and protein levels in the tumor tissues and paracancerous tissues was detected by real-time PCR and Western blot.The hilar cholangiocarcinoma paraffin-embedded specimens (n=103) were collected. The protein expression of BMP3 was determined by immunohistochemical method, and the relationship of BMP3 protein ex-pression with clinical pathological characteristics was evaluated.RESULTS:In the 30 patients with hilar cholangiocarcino-ma, the expressions of BMP3 protein and mRNA in 22 cases of tumor tissues were significantly decreased compared with the adjacent normal tissues.The results of immunohistochemistry showed that 87 cases were negative and 16 cases were weakly positive in all 103 cases of hilar cholangiocarcinoma.The expression of BMP3 protein was associated with the tumor TNM staging, lymph node metastasis and tumor differentiation (P<0.05).CONCLUSION:BMP3 gene might be inhibited in human hilar cholangiocarcinoma.The down-regulation of BMP3 gene might be associated with the carcinogenesis and devel-opment of hilar cholangiocarcinoma.

Objective To analyze the clinical value of serum and ascites tumor markers in the diagnosis of benign and malignant ascites ,and to provide reference for clinical diagnosis and treatment of ascites .Methods A total of 168 patients with ascites in Third Affiliated Hospital of Liaoning University of Chinese Medicine from November 2015 to June 2016 were selected .All patients were collected abdominal paracentesis ascites ,analysis of ascites tumor markers .Vein blood sample were collected ,centrifuged up-per serum and detected tumor markers including CA153 ,BXTM and CEA ,all patients underwent pathological test for diagnosis of benign and malignant ascites .The normal distribution method was used to determine the standard of judging malignant ascites and serum ,with the pathological diagnosis as the gold standard ,the four grid diagnosis table was used to calculate the sensitivity and specificity of different combination of ascites tumor markers and serum tumor markers in diagnosis of benign and malignant ascites . Results Among 168 cases ,56 cases were diagnosed with malignant ascites ,112 cases with benign ascites by pathological test .Com-bined detection results of three tumor markers in ascites :57 cases were malignant ,111 were benign .Combined detection results of three tumor markers in serum :64 cases were malignant ,104 cases were benign .The sensitivity and specificity of three tumor mark-ers in ascites were 85 .71% and 91 .96% respectively ,which were higher than 82 .41% and 83 .93% of the three tumor markers in serum .Conclusion The sensitivity and specificity of three tumor markers in ascites for diagnosis of benign and malignant ascites were superior ,which could be used as an important auxiliary means for diagnosis on the overall condition of patients with ascites , and provides an important reference for the subsequent selection other inspection methods .

Objective To study the effects of chronic manganism on hearing and cochlear cells in rats by using animal model of chronic manganism .Methods Sixty adult SD rats were randomly divided into Mn - exposed and controlgroups.RatsweretreatedwithMnCl24H2O(100mg·kg -1·d-1)ordeionizedwaterbygastricperfusion, lasted for 12 weeks .The Mn concentration in peripheral blood was measured respectively at 4 weeks ,8 weeks and 12 weeks after treatment .At 12 weeks after treatment ,the auditory brainstem response was recorded ,the hair cells morphology and counting were examined by stretched preparation of basilar membrane stained with FITC -phalloi-din ,and the spiral ganglion cells morphology and counting were studied by HE staining ,the ultrastructure changes of hair cells and spiral ganglion cells were detected by transmission electron microscopy .Results The blood Mn concentration increased gradually with time after treatment .ABR thresholds at 4 ,8 ,16 ,24 and 32 kHz were sig-nificantly increased at 12 weeks after treatment ,especially in the high-frequency range .Morphological study at 12 weeks after treatment showed loss of outer hair cells ,mainly in the basal turn of the cochlea ,and decreased number of spiral ganglion cells .The ultrastructure changes of outer hair cells and spiral ganglion cells included the break -ups ,disappearance or vacuolar change of mitochondria cristas .Conclusion Our data demonstrate that chronic man-ganism can cause loss of outer hair cells and spiral ganglion cells in cochlear in rats ,leading to hearing loss .

Objective To investigate the value of combined detection of alpha fetoprotein (AFP) ,gamma-glutamyl transferase (GGT)and Golgi glycoprotein (GP73)in the diagnosis of primary hepatocellular carcino-ma.Methods 109 cases of primary liver cancer treated in the hospital from June 2015 to June 2017 were en-rolled in the study as primary liver cancer group ,76 cases of liver cirrhosis were enrolled in the study as liver cirrhosis group and 70 cases of healthy people who underwent healthy assessment were enrolled in the study as control group.The peripheral venous blood in each group was extracted fasting in the morning ,and the ser-um was separated.The levels of GP73 were measured by enzyme linked immunosorbent assay.The level of AFP was measured by chemiluminescent immunoassay ,and the level of GGT was measured by rate method. Results The serum levels of AFP ,GGT and GP73 in the primary liver cancer group were higher than those in the liver cirrhosis group and the control group ,and the serum levels of AFP ,GGT and GP73 in the liver cir-rhosis group were higher than those in the control group ,and the differences were statistically significant (P＜0.05).The specificity and the sensitivity of the combined detection of AFP ,GGT and GP73 were higher than those of single detection of AFP ,GGT ,and GP73.Conclusion The combined detection of AFP ,GGT and GP73 has important value in the diagnosis of early primary liver cancer ,and has relatively high sensitivity and specificity ,which is worthy of further clinical study.

OBJECTIVETo observe the clinical effect of low-intensity ultrasound in promoting uterine involution following cesarean section.

METHODSA total of 122 women undergoing cesarean section were randomly selected and divided into low-intensity ultrasound treatment group (67 cases) and control group (55 cases). The women in the treatment group received daily low-intensity ultrasound treatment for 30 min 24 h after the delivery for 3 consecutive days, and the control group had no particular treatments. The descent of the uterine fundus and cessation of lochia was observed in the two groups.

RESULTSThe treatment group showed obviously greater descent of the uterine fundus with a higher rate of cessation of vaginal bleeding than the control group at 30 days postpartum. The ratio of incomplete uterine involution was significantly lower in the treatment group than in the control group.

CONCLUSIONLow-intensity ultrasound is effective in promoting uterine contraction and uterine involution following cesarean section.

Adult ; Cesarean Section ; rehabilitation ; Female ; Humans ; Postpartum Period ; Pregnancy ; Ultrasonic Therapy ; methods ; Ultrasonography ; Uterus ; diagnostic imaging ; Young Adult

Lactoferrin in milk is a multifunctional protein. In addition, lactoferrin has antiviral, antifungal and antiparasitic activity. In this study, the N-terminus from porcine lactoferrin (PLF-N) was designed to express the antimicrobial action of recombinant porcine lactoferrin. We cloned a 1077 bp fragment of the PLF gene from mammary gland tissue of the lactating sow at the third day. Comparing nucleotide sequence with four strains of PLF gene published on GenBank, the homology was more than 99%. With the reference template of the cloned fragment of PLF-N and optimizing codon bias, we synthesized the gene of N-terminus encoding porcine lactoferrin (PLF-NS). The high expression gene of PLF-NS was cloned into the fusion expression vector pET30b and expressed in E. coli BL21 (DE3). After induced with Isopropyl beta-D-1-Thiogalactopyranoside (IPTG), the target fusion protein was successfully expressed and identified in inclusion bodies by SDS-PAGE and Western blotting. The protein had a molecular weight of 42 kDa and accounted for 32% of the total cellular protein. After purification and renaturation, the purity of the expressed protein was 98%. The expressed PLF-NS protein showed obviously antibacterial activity. This method provides an excellent way for high expression of antimicrobial proteins when optimizing codon bias.
Amino Acid Sequence ; Animals ; Anti-Infective Agents ; metabolism ; pharmacology ; Base Sequence ; Escherichia coli ; genetics ; metabolism ; Genetic Vectors ; genetics ; Lactoferrin ; biosynthesis ; genetics ; pharmacology ; Molecular Sequence Data ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology ; Swine

Objective:To explore the real experience of developing self-consciousness, self-discovery, self-identity crisis and self-cognition improvement of young cancer patients from the initial stage of diagnosis, and to provide reference for medical staff to construct the intervention mode of psychosocial support.Methods:Using phenomenological method in qualitative research, 14 young cancer patients aged 18-25 years from April 2019 to August 2020 in Jiangsu Cancer Hospital were interviewed in semi-structured way separately at three time points: initial diagnosis, 6 months after diagnosis and 12 months after diagnosis. 7-step Colaizzi analysis was used to analyze the data.Results:Three themes were extracted as following. The dilemma of self-cognition, self-identity biases and the exploration of self-identity.Conclusions:Attention should be paid to the psychological problems of young cancer patients at all stages, and meet the needs of patients to talk, cooperate with professional psychotherapists, encourage patients to recognize themselves, tap the inner positive power, so as to improve their sense of self-identity.

ObjectiveTo explore the quality differences between steamed products and raw products of Citri Reticulatae Pericarpium（CRP）. MethodThe color of steamed products and raw products of CRP was determined from the perspective of appearance by electronic eye technique， and the quality differences between them was objectively characterized by the luminous value（L^*）， yellow-blue value（b^*）， red-green value（a^*） and total chromatic value（E^*_ab）. Based on this， ultra-high performance liquid chromatography（UPLC） was used to establish a fingerprint evaluation method with the mobile phase of acetonitrile（A）-0.1% formic acid aqueous solution（B） for gradient elution（0-5 min， 5%A； 5-30 min， 5%-20%A； 30-60 min， 20%-52%A）， detection wavelength at 270 nm， flow rate of 0.3 mL·min^-1 and column temperature of 30 ℃. The quality differences between steamed products and raw products of CRP were compared from the perspective of chemical composition， and correlation analysis was used to reveal the correlation between the difference in appearance color and the difference in internal chemical composition. ResultAfter being steamed， L^*， b^* and E^*_ab of CRP showed an overall decreasing trend， indicating that the color of the steamed products darkened and deepened from yellow to blue but still tended to be yellow， while a^* showed an overall increasing trend， indicating that the color of the steamed products tended to red. A total of 24 peaks were identified in the fingerprint profiles of raw products and steamed products of CRP， and 13 of the main peaks were identified. The precision， stability and repeatability studies showed that compared with the reference peak （peak 14， hesperidin）， the relative standard deviations（RSDs） of the relative peak area and relative retention time of the remaining peaks were<3.0%.The results of chemometric statistical analysis showed that there were some differences between raw products and steamed products of CRP， and 7 main differential components were identified， among which 5-hydroxymaltol（peak 1） and 5-hydroxymethylfurfural（peak 2） were the characteristic components of steamed products. The correlation analysis results showed that， in addition to the above two characteristic components， four components of peak 4， peak 10 （vicenin-2）， peak 23 （tangeretin） and peak 24 （5-demethylnobiletin） also correlated significantly with the color change （E^*_ab） of the samples （P<0.05， P<0.01）. ConclusionBefore and after steaming， not only the chemical composition changes， but also the color. Comparing the characteristic peaks of chemical composition difference and color difference before and after steaming of CRP， it is found that 5-hydroxymaltol， 5-hydroxymethylfurfural and peak 4 are common characteristic difference components， which can provide a reference for establishing the characteristic quality control method of steamed products， and quickly evaluating the quality difference between raw products and steamed products of CRP.