ObjectiveTo explore effect of Xianlian Jiedu prescription on the recurrence of colorectal cancer （CRC） and investigate the related mechanisms. MethodsA postoperative recurrence model was established in 25 Balb/c mice by injecting CT26 cells subcutaneously into the armpit， followed by surgical removal of 99% of the subcutaneous tumor. The mice were randomly divided into model group， low-dose Xianlian Jiedu prescription （XLJDP-L） group （6.45 g·kg^-1·d^-1）， medium-dose Xianlian Jiedu prescription （XLJDP-M） group （12.9 g·kg^-1·d^-1）， high-dose Xianlian Jiedu prescription （XLJDP-H） group （25.8 g·kg^-1·d^-1）， and 5-fluorouracil （5-FU） group （1×10^-3 g·kg^-1·d^-1）. The mice were euthanized after 14 days of continuous intervention， and recurrent tumor tissue was harvested. Hematoxylin and eosin （HE） staining was used to observe pathological and morphological changes in the recurrent tumor tissue. Immunohistochemistry （IHC） was employed to assess the expression of proliferating cell nuclear antigen （Ki67）， vascular endothelial growth factor （VEGF）， and platelet-endothelial cell adhesion molecule （CD31） in recurrent tumor tissue. The Western blot was used to detect the protein expression levels of angiopoietin-2 （ANG-2）， VEGF， phosphorylated-protein kinase B （p-Akt）， protein kinase B （Akt）， phosphorylated-phosphatidylinositol 3-kinase （p-PI3K）， and phosphatidylinositol 3-kinase （PI3K） in recurrent tumor tissue. ResultsBefore treatment， there were no statistical differences in tumor volume， tumor weight， and body mass among the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group compared to the model group， indicating model stability. After treatment， compared with those in the model group， the tumor volume and tumor weight in the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group were significantly reduced （P<0.01）， showing dose dependency. Meanwhile， there were no significant differences in body weight among the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group compared to the model group. HE staining showed that compared with that in the model group， tumor tissue in the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group had loosely arranged cells， increased intercellular spaces， small and shriveled nuclei， light staining， fewer mitotic figures and atypical nuclei， and increased necrotic areas. IHC showed that compared with those of the model group， the positive rates of Ki67， VEGF， and CD31 in the recurrent tumor tissue of the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group were significantly reduced （P<0.01） in a dose-dependent manner. Western blot results showed that compared with those of the model group， the protein expression levels of ANG-2 and VEGF in the recurrent tumor tissue of the XLJDP-L， XLJDP-M， and XLJDP-H groups and the 5-FU group were significantly downregulated （P<0.05， P<0.01）， and the p-Akt/Akt and p-PI3K/PI3K ratios were significantly decreased in a dose-dependent manner （P<0.05， P<0.01）. ConclusionXianlian Jiedu prescription significantly inhibits the recurrence of CRC in mice after subcutaneous tumor surgery. The mechanism may involve regulating the PI3K/Akt pathway and downregulating key angiogenic proteins such as ANG-2， VEGF， and CD31.

Objective Construct a human SNP genetic marker detection system using next generation sequencing technology and validate the system performance.Methods Based on multiple PCR amplification and capture technology,a set of detection system containing 621 autosomal SNPs and 398 Y chromosome SNPs was developed for accurate identification of individuals.To comprehensively evaluate the performance of the system,research was conducted on the overall coverage and balance,accuracy,sensitivity,repeatability,simulated degradation of samples,suitability of sample types,species specificity,mixed sample differentiation of the panel and so on.Results This system showed high coverage,uniformity and accuracy.when the input DNA was 0.062 5 ng,The system could detect more than 1 000 SNPs sites.And the system has a high detection efficiency for degradation test materials under the appropriate starting quantity.The repeatability between different personnel and different PCR instruments was 99.7%,indicating good repeatability.It is suitable for common forensic test materials,has high specificity,and has no spot detection for non-human samples in the test environment.When the gender ratio is 1∶20 or 20∶1,a single sample and mixed samples can still be distinguished.Conclusion The system has good classification accuracy and repeatability,with high sensitivity and high detection flux.It has strong detection ability for degraded and mixed samples,and can effectively eliminate interference from non-human species,improving individual recognition ability.

Background Cyclic AMP response element binding protein (CREB) and miR-132-3p have been proved to be related to many neurodegenerative diseases. Our research group previously has demostrated that the neurotoxicity of aluminum is relevant to abnormal phosphorylation of tau protein, but whether aluminum affects the abnormal phosphorylation of tau protein through GREB and miR-132-3p has not been reported yet . Objective To investigate the effect of aluminum on CREB and miR-132-3p during abnormal phosphorylation of tau protein in rat hippocampus. Methods Twenty-eight two-month-old SD rats with comparable weigh, were randomly assigned to four groups: control group (saline) and low, middle, and high dose exposure groups [10, 20, and 40 μmol·kg⁻¹ Al(mal)₃] with each group containing 7 rats, and the exposure period was 3 months by intraperitoneal injection every other day. After rats’ exposure to aluminum, Morris water maze was employed to assess their capabilities of learning and memory. The miR-132-3p gene expression level was detected by quantitative real-time PCR (qRT-PCR). The levels of CREB, phosphorylated CREB (p-CREB) (Ser133), RAS p21 protein activator 1 (RASA1) tau, and p-tau (Ser396) proteins were determined by Western blot. Results The results of Morris water maze showed that in the navigation experiment (from first day to the fifth day), the average escape latency of the rats exposed to three doses of aluminum was longer than that of the control rats (P<0.05). The middle dose group and the high dose group demonstrated shorter duration and lower frequency of platform traversal in the designated quadrant when compared to the control group and the low dose group (P<0.05). Moreover, the duration in the target quadrant of the rats exposed to high dose aluminum was shorter than that of the rats exposed to medium dose aluminum (P<0.05). The results of Morris water maze suggested that aluminum could damage the learning and memory ability of rats. The qRT-PCR findings indicated a decline in miR-132-3p gene expression in rat hippocampus correlating with higher Al(mal)₃ dose (P<0.05). The Western blot test showed that the protein expressions of CREB and p-CREB (Ser133) were reduced in both the middle dose group and the high dose group (P<0.05) when compared to the control group and the low dose group, and likewise, compared to the control group, the group receiving low dose exhibited lower level of p-CREB (Ser133) protein expression (P<0.05). It was found that the further increase of aluminum exposure dose would lead to the further decrease of CREB and p-CREB (Ser133) protein expression levels (F=36.429, P<0.001; F=78.672, P<0.001), aluminum exposure dose was negatively correlated with the expression levels of the two proteins (r=−0.848, P<0.001; r=−0.928, P<0.001). The expression levels of RASA1 protein and tau protein in the aluminum exposure groups surpassed those in the control group (P<0.05). The tau protein phosphorylation level was higher in the middle dose group than in the control group (P<0.05), while the high dose group showed elevated phosphorylation level relative to the control group, the low dose group, and the middle dose group (P<0.05). Conclusion Aluminum may promote abnormal phosphorylation of tau protein by affecting CREB and miR-132-3p, which eventually leads to the impairment of learning and memory ability.

BACKGROUND/OBJECTIVES: Chronic renal failure (CRF) is a complex pathological condition that lacks a cure. Certain Chinese medicines, such as melittin, a major component in bee venom, have shown efficacy in treating CRF patients. On the other hand, the mechanisms underlying the therapeutic effects of melittin are unclear.MATERIALS/METHODS: A 5/6 nephrectomy model (5/6 Nx) of renal failure was established on rats for in vivo assays, and mouse podocyte clone 5 (MPC5) mouse podocyte cells were treated with angiotensin II (AngII) to establish an in vitro podocyte damage model. The 24-h urine protein, serum creatinine, and blood urea nitrogen levels were evaluated after one, 2, and 4 weeks. Hematoxylin and eosin staining, Masson staining, and periodic acid-Schiff staining were used to examine the pathological changes in kidney tissues. A cell counting kit 8 assay was used to assess the cell viability. Reverse transcription polymerase chain reaction and Western blot were used to assess the mRNA and protein levels in the cells, respectively. RESULTS: In the rat 5/6 Nx, melittin reduced the 24-h urinary protein excretion and the serum creatinine and blood urea nitrogen levels. Furthermore, the renal pathology was improved in the melittin-treated 5/6 Nx rats. Melittin promoted podocin, nephrin, Beclin 1, and the LC3II/ LC3I ratio and inhibited phosphorylated mammalian target of rapamycin (mTOR)/mTOR in 5/6 Nx-induced rats and AngII-induced MPC5 mouse podocyte cells. Moreover, inhibiting autophagy with 3-MA weakened the effects of melittin on podocin, nephrin, and the LC3II/ LC3I ratio in podocytes. CONCLUSION Melittin may offer protection against kidney injury, probably by regulating podocyte autophagy. These results provide the theoretical basis for applying melittin in CRF therapy.

Objective:To observe any effects of contralateral repeated transcranial magnetic stimulation (rTMS) of the swallowing motor cortex on the swallowing and brainstem auditory evoked potentials (BAEPs) of stroke survivors with dysphagia.Methods:A total of 83 stroke survivors with dysphagia were randomly divided into an ipsilesional stimulation group ( n=22), a contralesional stimulation group ( n=21), a bilateral stimulation group ( n=20), and a control group ( n=20). In addition to their conventional dysphagia training, those in the three stimulation groups received 3Hz rTMS while the control group was given fake stimulation. The treatment was administered daily for 20 minutes, 6 days a week, for 5 consecutive weeks. Before and after the treatment, swallowing function was assessed videofluoroscopically and using the Dysphagia Outcome and Severity Scale (DOSS). The oral and pharyngeal stages of swallowing were evaluated using the videofluoroscopic dysphagia scale (VDS). Brain stem conduction was assessed using BAEPs. Results:After treatment the average DOSS scores of all 4 groups were significantly better than before the treatment. The average DOSS scores of the contralesional and bilateral sti-mulation groups were then significantly better than those of the other two groups. The sub-item and total VDS scores of all 4 groups had decreased significantly, but the average score of the bilateral stimulation group was significantly lower than the control group′s average. Ipsilesional stimulation significantly improved the VDS sub-item scores for the triggering of pharyngeal swallowing, laryngeal elevation, and pharyngeal transit time compared with the control group. In the contralesional stimulation group the average total score and the VDS sub-item scores for apraxia, premature bolus loss, oral transit times, the triggering of pharyngeal swallowing, vallecular residue, laryngeal elevation, coating on the pharyngeal wall, and pharyngeal transit time were significantly lower than those of the control group, on average. After the treatment the latencies of BAEP waves I, III and V and the I-III, III-V and I-V interpeak intervals had decreased significantly in all four groups, but the average latencies and intervals of the bilateral and contralesional groups were significantly shorter than those of the control group. The latencies and intervals of the bilateral stimulation group were then significantly shorter than those in the ipsilesional stimulation group on average. The average latency of wave V in the bilateral stimulation group (6.53±0.73ms) was significantly shorter than that in the contralesional stimulation group after the treatment.Conclusion:Bilateral rTMS over the swallowing motor cortex combined with conventional dysphagia training can significantly improve the swallowing of dysphagic stroke survivors.

Objective:To study the neurodevelopmental prognosis and risk factors for adverse outcomes of neonatal seizure.Methods:From December 2019 to November 2020, infants with neonatal seizure diagnosed in our hospital were enrolled in this retrospective study. Based on survival or not, mental development index (MDI), psychomotor development index (PDI) and seizure episodes at the age of 12 months, the infants were assigned into adverse outcome group and normal outcome group. The risk factors for adverse outcomes were statistically analyzed.Results:A total of 75 infants were enrolled,including 39 cases in adverse outcome group and 36 in normal outcome group. 69 cases showed abnormal amplitude-integrated electroencephalogram(aEEG), including 38 mildly abnormal cases,23 moderately abnormal cases and 8 severely abnormal cases, The incidences of adverse outcomes and mortality rates were significantly different ( P<0.05) among infants with different severity levels of aEEG abnormalities and the severity levels of aEEG abnormalities were positively correlated with adverse outcomes ( r=0.367, 0.471, P<0.05).Univariate analysis showed that adverse outcome group had significantly higher incidences of chorioamnionitis, seizure onset age ≤3 d, 5 min Apgar score ≤3, cranial ultrasound abnormalities, brain MR abnormalities and aEEG abnormalities than normal outcome group ( P<0.05).Logistic regression analysis showed that seizure onset age ≤3 d ( OR=3.988, 95% CI 1.376-11.674), abnormal brain MR ( OR=3.296, 95% CI 2.383-17.377) and bilirubin encephalopathy ( OR=3.792,95% CI 2.110-13.216) were independent risk factors for adverse outcomes of neonatal seizure. Conclusions:For neonatal seizure, the infants with more severe abnormal aEEG will have higher incidences of adverse outcomes and mortality. Seizure onset age ≤3 d, brain MR abnormalities and bilirubin encephalopathy were independent risk factors for adverse outcomes of neonatal seizure.

Objective:To study the complications and prognosis of extremely premature infants(EPIs) with gestational age (GA) <28 w.Methods:From January 2016 to December 2020, EPIs with GA <28 w admitted to NICU of our hospital were retrospectively studied. Clinical data of the infants and their mothers during pregnancy were reviewed. According to the prognosis, the infants were assigned into the survival group, the death group and the withdrawal group. According to GA, the infants were assigned into <26 w goup, 26~26 +6 w group, ≥27 w group. According to birth weight (BW), the infants were assigned in to ≤750 g group, 750~999 g group and ≥1 000 g group. SPSS 26.0 was used for data analysis. Results:A total of 265 EPIs were included, 122 (46.0%) in the survival group, 47 (17.7%) in the death group and 96 (36.2%) in the withdrawal group. GA and BW of the survival group were significantly higher than the death group and the withdrawal group ( P<0.05). The incidences of tracheal intubation (92.2%) and pulmonary hemorrhage (42.2%) in the death group were the highest among the three groups. The survival group had the highest application of prenatal glucocorticoids (80.3%) and pulmonary surfactants (99.2%) ( P<0.05). In the survival group, the top 3 common complications were bronchopulmonary dysplasia (BPD) (68.0%), pulmonary infections (65.6%) and neonatal sepsis (34.4%). The survival rate increased with GA and BW. Conclusions:The survival rate of EPIs is closely correlated with GA and BW. EPIs with pulmonary hemorrhage and tracheal intubation have poor prognosis. Prenatal glucocorticoids and pulmonary surfactant may improve clinical outcome. BPD and pulmonary infections are common complications of surviving EPIs.

Objective:To study the pathogen profile and clinical characteristics of catheter-related bloodstream infection (CRBSI) in very low birth weight infants (VLBWI) receiving peripherally inserted central venous catheter (PICC).Methods:From November 2018 to October 2021, VLBWI with CRBSI after PICC admitted to neonatal intensive care unit (NICU) of our hospital were retrospectively studied. The pathogen profile and drug resistance characteristics were analyze. The infants with bacterial infection were assigned into gram-negative (G -) group and gram-positive (G +) group. Their general status, PICC duration and the site of PICC, body weight at infection, use of lipid emulsion and prophylactic antibiotics, clinical manifestations and laboratory results were compared between the two groups. Results:A total of 70 infants with CRBSI were included and 70 strains of pathogenic bacteria were detected. 50.0% (35/70) were G - bacteria with klebsiella pneumoniae (34.3%, 12/35) and serratia marcescens (34.3%, 12/35) as the most common bacteria. Klebsiella pneumoniae showed more severe drug resistance. 47.1% (33/70) were G + bacteria and the most common strain was staphylococcus epidermidis (45.5%, 15/33) with a majority of methicillin-resistant (86.7%, 13/15). 2 cases (6.1%, 2/33) had bacillus cereus infection and both suffered quick death. 2.9% (2/70) were fungi infection. The main clinical manifestation of CRBSI in VLBWI was apnea and shock was the most common complication. G + group showed significantly higher gestational age and lipid emulsion usage but lower body weight than G - bacteria group. No significant differences existed in clinical manifestations, laboratory results and prognosis between the two groups. Conclusions:Most pathogens causing CRBSI in VLBWI with PICC are opportunistic pathogens. It is difficult to differentiate G + and G - bacterial infection based on clinical manifestations and laboratory results. However, VLBWI with higher gestational age and lipid emulsion usage but lower body weight are more susceptible to G + bacterial infection.

ObjectiveTo investigate the mechanism by which Shenbai Jiedu prescription （SBJDF） inhibits the proliferation of colorectal cancer （CRC） HCT116 cells. MethodAfter 48 h treatment of HCT116 cells with SBJDF （0， 0.25， 0.5， 1， 2， 4 g·L^-1）， the viability of HCT116 cells were determined by methyl thiazolyl tetrazolium （MTT） colorimetry. Following the classification of cells into blank control group and SBJDF （1， 2， 4 g·L^-1） groups， the effect of SBJDF on HCT116 cell morphology was observed under an inverted microscope. The effects of SBJDF on the proliferation of HCT116 cells and mitochondrial membrane potential （Δψm） were detected by colony formation assay and JC-1 probe， respectively. The flow cytometry was then performed for determining cell cycle distribution and apoptosis. The effects of SBJDF on cell cycle-， apoptosis-， and nuclear factor kappa-B （NF-κB） signaling pathway-related proteins were determined by Western blot. ResultSBJDF effectively inhibited the vitality of HCT116 cells and changed their morphology in a concentration-dependent manner. Compared with the blank control group， SBJDF at 1， 2， 4 g·L^-1 significantly reduced cell colony formation （P<0.05， P<0.01），and SBJDF at 2 and 4 g·L^-1 arrested the HCT116 cell cycle at G₀/G₁ phase （P<0.05， P<0.01）. Compared with the blank control group， SBJDF at 1， 2， 4 g·L^-1 remarkably down-regulated the protein expression of CyclinD₁ （P<0.05， P<0.01）. SBJDF at 2 and 4 g·L^-1 lowered the CyclinA₂ and cyclin-dependent kinase 4 （CDK4） （P<0.05， P<0.01）. SBJDF at 4 g·L^-1 reduced the cyclin-dependent kinase 1 （CDK1） （P<0.01）. Compared with the blank control group， SBJDF at 2 and 4 g·L^-1 induced HCT116 cell apoptosis， down-regulated the protein expression of anti-apoptosis-related proteins Bcl-2 and Bcl-xl as well as the NF-κB signaling pathway-related proteins IκB kinase α （IKKα），inhibitor α of NF-κB （IκBα），and phospho-NF-κB p65 （p-p65） （P<0.05， P<0.01）， and diminished the mitochondrial membrane potential of HCT116 cells. ConclusionSBJDF inhibits the proliferation of HCT116 cells， which may be related to its inhibition of the activation of NF-κB signaling pathway and the induction of cell cycle arrest and apoptosis.

ObjectiveTo study the mechanism of Shenbai Jiedu prescription inhibiting the proliferation of HCT116 colorectal cancer （CRC） cells by regulating the phosphatase and tensin homolog deleted on chromosome ten （PTEN）/phosphatidylinositol 3-kinase （PI3K）/ protein kinase B （Akt） signaling pathway. MethodShenbai Jiedu prescription was extracted by water extraction and alcohol precipitation to prepare freeze-dried powder. HCT116 cells were cultured in vitro， and treated with different concentrations of Shenbai Jiedu prescription （2， 4， 8， 16 g·L^-1）. The inhibitory effect of Shenbai Jiedu prescription on the proliferation of HCT116 cells was tested by methyl thiazolyl tetrazolium （MTT）. Real-time quantitative PCR was used to detect the mRNA expression levels of PTEN， PI3K， Akt， glycogen synthase kinase-3β （GSK-3β）， c-Myc， survivin and Cyclin D₁. Western blot was employed to measure the protein expression levels of PTEN， phosphorylated PTEN （p-PTEN）， PI3K， Akt， phosphorylated Akt （p-Akt）， GSK-3β， phosphorylated GSK-3β （p-GSK-3β）， c-Myc， survivin and Cyclin D₁， β-catenin nuclear import was explored by immunofluorescence assay. ResultCompared with the control group， Shenbai Jiedu prescription inhibited the proliferation of HCT116 cells in a dose-dependent manner （P<0.01）. Compared with the control group， the mRNA expression levels of PTEN and GSK-3β were up-regulated whereas those of PI3K， Akt， c-Myc， survivin and CyclinD₁ were down-regulated after treatment with Shenbai Jiedu prescription （P<0.01）. The protein expression levels of PTEN， p-PTEN and GSK-3β were up-regulated whereas those of PI3K， Akt， p-Akt， GSK-3β， p-GSK-3β， c-Myc， survivin and CyclinD₁ were down-regulated （P<0.05， P<0.01）. Immunofluorescence assay showed that Shenbai Jiedu prescription suppressed β-catenin nuclear import in HCT116 cells. ConclusionShenbai Jiedu prescription inhibited the proliferation of HCT116 cells via the mechanism of regulating the PTEN/PI3K/Akt signaling pathway.