Aim To investigate the role of locus coeruleus(LC)-norepinephrinergic system and sympathetic nervous system in immunosuppression under cold stress, using 6-hydroxydopamine(6-OHDA) as chemical sympathectomy.Methods Rats were maintained in cold chamber at 4℃ for 4 h.The 51Cr release assay from YAC-1 cells was used to determine the splenic NK cell activity, the double staining of ABC method was employed to observe the immunoreactive expression of Fos, arginine-vasopressin(AVP), oxytocin(OT) and tyrosine hydroxylase(TH), and conventional radioimmunoassay was used to measure plasma corticosterone (CORT) concentrations.Results Central sympathectomy with intracerebroventricular(i.c.v.) injection of 6-OHDA r educed significantly the elevation of plasma corticosterone level, the expressio n of Fos in hypothalamic paraventricular nucleus(PVN) and in locus coeruleus(LC ), as well as the suppression of NK activity induced by cold stress at 4℃ for 4 h. Peripheral sympathectomy with intraperitoneal (i.p.) injection of 6-OHDA al so reversed the cold stress-induced suppression of NK cytotoxicity, but without significant effect on Fos expression in brain. Double staining showed that amon g the Fos-positive neurons in PVN only a few co-expressed Fos and AVP or Fos a nd OT, while in LC the majority of Fos-positive neurons co-expressed Fos and T H.Conclusion The mechanism of suppression of NK activity induce d by cold stress may be mediated through HPA axis activated partially by central LC-norepinephrinergic system and through the peripheral sympathetic nervous system.

Objective To investigate the clinical value of miRNA-34a,miRNA-34c,and miRNA-135a in the early diagnosis of mild cognitive impairment (MCI) and Alzheimer's disease (AD).Methods The patients were collected in the outpatient and inpatient of Neurology during Aug 2014 to May 2016.The levels of miRNA-34a,miRNA-34c,an dmiRNA-135a were detected with quantitative real-time polymerase chain reaction (qRT-PCR).Results AD patients had higher level of miRNA-34a than the controls.The levels of miRNA-34c were higher in MCI and AD patients,while lower levels of miRNA-135a compared to the controls.Conclusions The miRNA 34a and miRNA-135a were likely to became the biomarker in early diagnosis of MCI and AD.

Objective To establish the HepG2 cell lines which can stably express and replicate hepatitis 13 virus (HBV).Methods One point two X unit length of HBV genome was cloned intn SalⅠsite of the eukaryotic expression vector pREP10 to construct the recombinant plasmid pREP-HBV. Human hepatoblastoma cell HepG2 was transfected with pREP-HBV by Lipofectamine 2000 and seh,cted by bygromycin at the concentration of 250?g/mL.HBsAg and HBeAg were monitored by enzyme linked immunosorbent assay (ELISA)kits.H13V particles presemed in supernatant were ex- amincd by electronic microscopy.DNA isolated from intracellular HBV core particles was analyzed by Southbern blot using HBV-specific probe.Results The recombinant vector pREP HBV containing 1.2?unit length of HBV DNA was constructed successfully.After transfection of pREP-HBV to HepG2 cells and consistently cultured in hygromycin selective medium.5 drug-resistant cell lines, RHBV1-RHBV5.were established,and all of them could stably express HBsAgand HBeAg.South ern blot analysis revealed that HBV could replicate in all cell lines,as confirmed by the presence of replicateintermediatc DNA in intracellular HBV core particles.Clustered 42 nm Dane particles as well as 22-26 nm spherical H13sAg particles in condensed cuhure supernatant were visualized by elec tronic microsopic analysis.Conclusion HepG2 ceil lines in which HBV can replicate and express specific antigens are successfully established.Up to now,the cells have been passaged every three days for 50 times.

Objective:To explore and analyze the characteristics and causes of intensive nursing needs of COVID-19 patients.Methods:This was a cross-sectional study. Critical patients′ treatment and care information were extracted through the HIS system of a designated hospital from critical patients with COVID-19. We used statistical descriptions to analyze patient general information, nursing degree, and therapeutic intensive care needs. The chi-square test or Fisher′s exact test was used to analyze the differences between characteristics of patients and nursing degree.Results:A total of 1 849 cases of critical patients were collected, including special grade nursing care 697(37.70%), grade I nursing care(62.30%). There were 893(48.30%)patients aged≥65 years, and 853(46.13%)patients with one or more complications. All patients required the nursing routine of infection department, and the therapeutic intensive care followed was vein blood collection(99.62%), oxygen inhalation(98.92%), oral medication(98.65%), finger pulse oxygen monitoring(77.07%)and pharyngeal swab collection(71.93%). The nursing degree of the patients with different gender, age, admission mode, body position and complications were statistically significant( P<0.05). Conclusions:The intensive care nursing needs of COVID-19 patients are complex, which are special in treatment, life and psychology, and the tasks of nursing are heavy, difficult and professional. Special attention should be paid to elderly patients and patients with complications. Our hospital has established a provisional nursing department during the COVID-19 outbreak, coordinated nursing manpower and quality of nursing management, and set up a nursing expert consultation team to further enhance the quality of nursing care for critical COVID-19 patients.

Objective:To compare the clinical value of stent assisted intestinal bypass and temporary loop ileostomy in laparoscopic low anterior resection of rectal cancer.Method:In this retrospective analysis, 57 patients undergoing laparoscopic low anterior resection for rectal cancer in the First Affiliated Hospital of Ningbo University from Jan 2020 to Jan 2022 were divided into intestinal bypass group (36 cases) and loop ileostomy group (21 cases).Result:There were no significant differences in postoperative GI function recovery and postoperative complication rate between the two groups (all P>0.05). The levels of albumin, prealbumin and hemoglobin in the intestinal bypass group were better than those in the ileostomy group when evaluated on 3rd months after operation [(40.5±2.3) g/L vs. (38.1±2.6)g/L、(26.4±2.7)mg/dl vs. (24.5±2.0)mg/dl、(137.6±5.9) g/L vs. (134.0±7.0) g/L, t=3.605、2.743、2.085, all P<0.05]. Hospital expenses of the intestinal bypass group was lower [(571 000±7 500) yuan vs. (69 300±9 100) yuan, t=-5.477, P<0.05]. Conclusion:Compared with traditional ileostomy, the stent assisted intestinal bypass reduces trauma with lower expenses and improves patients' status after laparoscopic low anterior resection for rectal cancer.

Objective To investigate the therapeutic effect of Shugan Heluo Xingpi Decoction on CCl 4 -induced liver fibrosis in rats, and the underlying molecular mechanism. Methods Sixty male SPF Wistar rats were randomly assigned to six groups: blank control, model, positive control, high, medium or low dose groups of Shugan Heluo Xingpi Decoction ( n =10 per group). The liver fibrosis rat model was induced by an intraperitoneal injection of 40% CCl 4 oil solution. Rats in the blank control and model groups were administered 10 mL/kg normal saline by gavage, rats in the positive control group were administered 50 mg/kg silibinin meglumine by gavage, while rats in high-dose, medium-dose and low-dose groups of Shugan Heluo Xingpi Decoction were administered 12.42 g/kg, 6.21 g/kg and 3.11 g/kg (crude drug/body weight) by gavage, respectively, daily for 8 weeks. Rats was sacrificed after 8 weeks, during which the physiological status of rats in each group was dynamically monitored. Following sacrifice, serum was collected to detect HYP using alkaline hydrolysis colorimetry and the expression levels of AST, ALT, total protein (TP), and Alb using an automatic biochemical analyzer. The pathological morphological changes in the liver were detected by H & E staining and Masson staining, and the mRNA and protein levels of Wnt1, β-Catenin and Cyclin D1 were detected by RT-qPCR and Western Blot. Measurement data were compared across groups using one-way ANOVA with post-hoc LSD- t test. Results Compared with the model group, after silibinin meglumine and Shugan Heluo Xingpi Decoction intervention, the physiological status of rats was significantly improved; serum levels of HYP, ALT, AST and Glo were significantly decreased, while serum levels of TP, Alb and A/G were significantly increased (all P < 0.05). Compared with the positive control group, serum levels of ALT and AST were significantly increased (all P < 0.05), while the levels of TP, Alb and A/G were significantly decreased (all P < 0.05) in low-dose group of Shugan Heluo Xingpi Decoction. H&E staining showed mild portal vein fibrosis with a few fibrous septa and mild steatosis of hepatocytes in the positive control group, obvious portal vein fibrosis with a few fiber septum in the low dose group, a few portal vein fibrosis in the medium dose group, while no obvious abnormality in the high dose group of Shugan Heluo Xingpi Decoction. Masson staining revealed that the therapeutic effect of high dose group of Shugan Heluo Xingpi Decoction on collagen deposition was superior to silibinin meglumine and medium and low dose of Shugan Heluo Xingpi Decoction (all P < 0.05), and was generally equivalent to high dose of Shugan Heluo Xingpi Decoction. Silibinin meglumine and medium and high doses of Shugan Heluo Xingpi Decoction inhibited more significantly the mRNA and protein expression of Wnt1, β-catenin and Cyclin D1 (all P < 0.05). Conclusion Shugan Heluo Xingpi Decoction shows anti-hepatic fibrosis effect, with a greater effect at higher doses. Regulating Wnt/β-Catenin signaling pathway may be one of the underlying molecular mechanisms.

The many-banded krait, Bungarus multicinctus, has been recorded as the animal resource of JinQianBaiHuaShe in the Chinese Pharmacopoeia. Characterization of its venoms classified chief phyla of modern animal neurotoxins. However, the evolutionary origin and diversification of its neurotoxins as well as biosynthesis of its active compounds remain largely unknown due to the lack of its high-quality genome. Here, we present the 1.58 Gbp genome of B. multicinctus assembled into 18 chromosomes with contig/scaffold N50 of 7.53 Mbp/149.8 Mbp. Major bungarotoxin-coding genes were clustered within genome by family and found to be associated with ancient local duplications. The truncation of glycosylphosphatidylinositol anchor in the 3'-terminal of a LY6E paralog released modern three-finger toxins (3FTxs) from membrane tethering before the Colubroidea divergence. Subsequent expansion and mutations diversified and recruited these 3FTxs. After the cobra/krait divergence, the modern unit-B of β-bungarotoxin emerged with an extra cysteine residue. A subsequent point substitution in unit-A enabled the β-bungarotoxin covalent linkage. The B. multicinctus gene expression, chromatin topological organization, and histone modification characteristics were featured by transcriptome, proteome, chromatin conformation capture sequencing, and ChIP-seq. The results highlighted that venom production was under a sophisticated regulation. Our findings provide new insights into snake neurotoxin research, meanwhile will facilitate antivenom development, toxin-driven drug discovery and the quality control of JinQianBaiHuaShe.