Separation of bufadienolides from Helleborus thibetanus Franch. by a combination approach involving macroporous resin column chromatography and gradient countercurrent chromatography.

In this study, a combination approach involving macroporous resin (MR) column chromatography and gradient countercurrent chromatography (CCC) was employed to enrich and purify bufadienolides from the roots and rhizomes of Helleborus thibetanus Franch. Initially, a D101 MR-packed column chromatography was utilized for fractionation and enrichment of the bufadienolides, which were effectively eluted from the column using a 60% ethanol solution. CCC was subsequently introduced to separate the enriched product using the ethyl acetate/n-butanol/water (EBuWat, 4:1:5, v/v) and EBuWat (5:0:5, v/v) solvent systems in a gradient elution mode. As results, five bufadienolides, including 6.1 mg of hellebrigenin-3-O-ß-D-glucoside (1), 2.2 mg of tigencaoside A (2), 8.3 mg of deglucohellebrin (3), 3.5 mg of 14 ß-hydroxy-3ß-[ß-D-glucopyranosyl-(1→6)-(ß-D-glucopyranosyl)oxy]-5α-bufa-20,22-dienolide (4), and 3.0 mg of 14ß-hydroxy-3ß-[(ß-D-glucopyranosyl)oxy]-5α-bufa-20,22-dienolide (5), were effectively separated from 300 mg of the enriched product. The respective high-performance liquid chromatography purities were as follows: 95.2%, 75.8%, 85.7%, 82.3%, and 92.8%. This study provides valuable insights for the efficient enrichment and separation of bufadienolides from Helleborus thibetanus Franch.

Bioassay-guided separation and identification of the anticancer composition from Curcuma longa L. by the combination strategy of methanol gradient countercurrent chromatography and ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry.

In this study, a combined strategy with methanol gradient countercurrent chromatography and ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry was introduced for the fractionation and identification of active constituents from Curcuma longa L. The gradient countercurrent chromatography separation was performed using the heptane-ethyl acetate-methanol-water (5:5:2:8, v/v) solvent system, in which the lower phase and methanol were used as the mobile phases. Constituents of turmeric with large partition coefficients were well resolved. Subsequent cytotoxicity analysis showed that the fractions 10, 11, 12, and 15 expressed significantly higher cytotoxicity against B16 mouse melanoma than the other fractions. Four compounds with potent activity, curcumin, demethoxycurcumin, and bisdemethoxycurcumin from fraction 11 and galanal A from fraction 15, were purified, and the half-maximal inhibitory concentrations were 18.5 ± 1.3, 7.8 ± 0.4, 20.4 ± 1.3, and 14.1 ± 0.8 µM, respectively. Ultra-high-performance liquid chromatography coupled with high-resolution mass spectrometry was then applied for compound identification from fraction 10 with constituents in very low content. A total of 14 diarylheptanoids were identified, which are supposed to be cytotoxic constituents. It proved that the strategy based on the combination of bioassay-guided methanol gradient countercurrent chromatography separation and ultra-high-performance liquid chromatography coupled with tandem mass spectrometry-assisted peak identification could be an efficient method for natural product screening and new drug discovery.

Use of Enhanced Recovery After Surgery (ERAS) in Laparoscopic Cholecystectomy (LC) Combined with Laparoscopic Common Bile Duct Exploration (LCBDE): A Cohort Study.

BACKGROUND The have been few reports on use of ERAS in LC combined with LCBDE to promote postoperative recovery of patients. Therefore, the purpose of this cohort study was to explore the use of ERAS in patients who underwent LC combined with LCBDE. MATERIAL AND METHODS We collected clinical data of 445 patients who underwent elective laparoscopic cholecystectomy combined with laparoscopic common bile duct exploration from January 2015 to February 2019 in our hospital and divided the patients into an E-LC group and an LC group. The stress response index, postoperative complication rate, and postoperative rehabilitation effect of the 2 groups were compared and analyzed. RESULTS The WBC count and CRP levels in the E-LC group were significantly lower than those of the LC group 1 day after surgery (p<0.05). In terms of the postoperative complications, the incidence of nausea, incisional pain, and vomiting in the E-LC group were lower than in the LC group, and the differences were statistically significant (p<0.05). In terms of the postoperative rehabilitation efficacy, flatus time and length of hospital stay after surgery in the E-LC group were significantly shorter than those in the LC group (p<0.05). CONCLUSIONS Use of ERAS in the perioperative period in patients who underwent LC combined with LCBDE reduces the stress response and postoperative complications and accelerates postoperative rehabilitation.

The complete mitochondrial genome of Sigara lateralis (Leach, 1817) (Nepomorpha: Corixidae).

The Sigara lateralis (Leach, 1817) is a small aquatic insect belonging to the family Corixidae. The study aims to reveal the genomic data of the mitochondrial genome of S. lateralis. The length of its circular mitochondrial genome is 15,725 bp long with an A + T bias (75.96%). The mitogenome comprises 37 genes, including 13 protein-coding genes (PCGs), 22 transfer RNA genes, and two rRNA genes. The phylogenetic analyses showed that the S. lateralis is the closest to S. septemlineata. These findings will help the conservation of Corixidae from the perspective of genetic evolution.

Exploring the Mechanism of Acupuncture's Influence on the Protease Activity and Free Radical Damage in Synovial Fluid of Rheumatoid Arthritis Rats Induced by Type II Collagen from the Release of Active Oxygen.

Acupuncture was studied to investigate the mechanism of its effect on protease vitality and free radical damage in Type I CIA rats induced by type II collagen. The study divided rats into a control group (injected with physiological saline, n = 10), a model group (injected with type II collagen, n = 10), and an intervention group (injected with type II collagen + acupuncture ST36 and GB39, 3 times a week, for a total of 4 weeks, n = 10) based on the different injected drugs. Then, various indicators of the mice were experimentally tested using joint index scoring, H&E histological staining, protein blotting, and immunohistochemistry staining methods. Acupuncture ST36 and GB39 can reduce arthritis scores, histological staining scores, and increase MVD in CIA rats. And reduce protease levels, alleviate inflammation, synovial hyperplasia, and angiogenesis. In addition, the intervention group TNF-α, IL-1ß and IL-6 mRNA were reduced, and the clearance rates of hydrogen peroxide free radicals and nitric oxide free radicals were increased. The expression levels of ROS and MDA decrease, while the expression levels of SOD increase It has been proved that acupuncture at ST36 and GB39 can inhibit the release of ROS, reduce protease activity, inflammation, synovial hyperplasia, angiogenesis and free radical damage, thus reducing the severity of CIA (Collagen-Induced Arthritis) in rats.

Multi-dimensional preparation of Thymus quinquecostatus Celak. by normal-phase flash chromatography coupled to counter-current chromatography.

In this study, a multi-dimensional chromatography system was developed by integrating normal-phase flash chromatography and counter-current chromatography to isolate flavonoids, phenylpropanoids, and thymol from the aerial parts of Thymus quinquecostatus Celak. In the online multi-dimensional switching system, a normal-phase flash chromatograph packed with 1.2 g of dry homogeneous silica gel mixture (containing 600 mg of methanol extract) was connected to counter-current chromatography via a six-port valve. Two two-dimensional separations were performed using n-heptane-ethyl acetate-methanol-water (6:4:6:4, v/v) and ethyl acetate-water solvent systems sequentially to separate the constituents of Thymus quinquecostatus Celak. The upper phase of the former solvent system was utilized as both elution solvent for flash chromatography and the stationary phase for counter-current chromatography, while the lower phase of the latter solvent system containing 10 mM trifluoroacetic acid was employed as elution solvent for flash chromatography and one mobile phase in pH gradient counter-current chromatography. Thymol (7) and xanthomicrol (8), two hydrophobic ingredients, were purified in the initial two-dimensional separation. The subsequent two-dimensional separation yielded six hydrophilic compounds, namely dihydrokaempferol-7-O-D-glucopyranoside (1), lithospermic acid (2), luteolin-7-O-glucuronide (3), rosmarinic acid (4), messerschmidin (5) and apigenin-7-O-D-glucuronide (6). This study represents the first documented use of online multi-dimensional normal-phase flash chromatography coupled to counter-current chromatography for separating constituents from Thymus quinquecostatus Celak.

Establishment and Verification of a Predictive Nomogram for New Vertebral Compression Fracture Occurring after Bone Cement Injection in Middle-Aged and Elderly Patients with Vertebral Compression Fracture.

OBJECTIVE: New vertebral compression fracture (NVCF) occurring after bone cement injection in middle-aged and elderly patients with vertebral compression fracture is very common. Preoperative baseline characteristics and surgical treatment parameters have been widely studied as a risk factor, but the importance of the patients' laboratory indicators has not been thoroughly explored. We aimed to explore the relationship between laboratory indicators and NVCF, and attempt to construct a clinical prediction model of NVCF together with other risk factors. METHODS: Retrospective analysis was performed for 200 patients who underwent bone cement injection (percutaneous kyphoplasty or vertebroplasty) for vertebral compression fractures between January 2019 and January 2020. We consulted the relevant literature and collated the factors affecting the occurrence of NVCF. Feature selection of patients with NVCF was optimized using the least absolute shrinkage and selection operator regression model, which was used to conduct multivariable logistic regression analysis, to create a predictive model incorporating the selected features. The discrimination, calibration, and clinical feasibility of the predictive model were assessed using the concordance index (C-index), calibration plots, and decision curve analysis. Internal validation was performed using Bootstrap resampling verification. RESULTS: Time from injury to surgery exceeding 7 days, low osteocalcin levels, elevated homocysteine levels, osteoporosis, mode of operation (percutaneous vertebroplasty), lack of postoperative anti-osteoporosis treatment, and poor diffusion of bone cement were independent risk factors for NVCF in middle-aged and elderly patients with vertebral compression fracture after bone cement injection. The C-index of the nomogram constructed using these seven factors was 0.895, indicating good discriminatory ability. The calibration plot showed that the model was well calibrated. Bootstrap resampling verification yielded a significant C-index of 0.866. Decision curve analysis demonstrated that the greatest clinical net benefit for predicting NVCF after bone cement injection could be achieved with a threshold of 1%-91%. CONCLUSION: This nomogram can effectively predict NVCF incidence after bone cement injection in middle-aged and elderly patients with vertebral compression fracture, thus aiding clinical decision-making and postoperative management, promoting effective postoperative rehabilitation, and improving the quality of life.

Exploring the mechanism of Astragali radix for promoting osteogenic differentiation based on network pharmacology, molecular docking, and experimental validation.

The present study used network pharmacology and molecular docking to predict the active ingredients and mechanisms of action of Astragalus radix (AR) to promote osteogenic differentiation of bone marrow mesenchymal stem cells (BM-MSCs), and cell experiments were conducted for verification. First, network pharmacology was used to predict the effective components, targets, and mechanisms of action of AR to promote osteogenic differentiation. The effective components and corresponding target proteins of AR, and the target proteins of osteogenic differentiation were collected through the database. The intersection targets of the two were used for the construction and analysis of a protein-protein interaction (PPI) network. Gene Oncology (GO) and Kyoto Encyclopedia of Genes, and Genomes (KEGG) enrichment analyses were conducted. Next, molecular docking technology was carried out to verify the interaction between the active ingredient and the target protein, and to select the appropriate effective active ingredient. Finally, the results of network pharmacology analysis were verified by in vitro experiments. A total of 95 potential targets were retrieved by searching the intersection of AR and osteogenic differentiation targets. PPI network analysis indicated that RAC-α-serine-threonine-protein kinase (Akt1) was considered to be the most reliable target for AR to regulate osteogenic differentiation. GO enrichment analysis included 21 biological processes, 21 cellular components and 100 molecular functions. KEGG enrichment analysis indicated that the class I phosphatidylinositol-3 kinase (PI3K)-serine-threonine kinase (Akt) signaling pathway may play an important role in promoting osteogenic differentiation. The results of molecular docking showed that quercetin's performance was improved compared with that of kaempferol. In vitro experiments showed that quercetin promoted the expression of osteogenic marker proteins (including collagen I, Runt-related transcription factor 2 and osteopontin) in BMSCs and activated the PI3K/Akt signaling pathway. AR acted on Akt1 targets through its main active component quercetin, and promoted the osteogenic differentiation of BM-MSCs by activating the PI3K/Akt signaling pathway.

Overexpression of maize GOLDEN2 in rice and maize calli improves regeneration by activating chloroplast development.

Golden2 (G2), a member of the GARP transcription factor superfamily, regulates several biological processes and phytohormone signaling pathways in plants. In this study, we used a rice codon-optimized maize G2 gene (rZmG2) to improve the regeneration efficiency of rice and maize calli for genetic transformation. We isolated a promoter driving strong and callus-specific expression from rice to drive rZmG2 transcription from a transgene after transformation of two indica and two japonica rice cultivars. The resulting rZmG2 transgenic calli turned green in advance at the differentiation stage, thus significantly raising the regeneration rates of the transgenic indica and japonica rice plants relative to control transformations. Similar effect of this gene on improving maize transformation was also observed. Transcriptome sequencing and RT-qPCR analyses showed that many rice genes related to chloroplast development and phytohormones are upregulated in rZmG2-transgenic calli. These results demonstrate that rZmG2 can promote embryogenic callus differentiation and improve regeneration efficiency by activating chloroplast development and phytohormone pathways. We also established a heat-inducible Cre/loxP-based gene-excision system to remove rZmG2 and the antibiotic selectable gene after obtaining the transgenic plants. This study provides a useful tool for functional genomics work and biotechnology in plants.

A Neglected Case of Colonic Tuberculosis With Thoracolumbar Enterocutaneous Fecal Fistulae.

Although intestinal tuberculosis (ITB) typically affects ileocecal segments, the complication of tubercular enterocutaneous fistula is very rare. As an isolated phenomenon, primary intestinal manifestation without extraintestinal tuberculosis (TB) is exceptional and rarely reported. We present a patient with isolated ITB with six spontaneous thoracolumbar tubercular enterocutaneous fistulae. A 37-year-old Chinese woman was admitted to our institution complaining of escape of fecal matter through several openings in her back over the previous 20 years. She had nonspecific abdominal symptoms (occasional abdominal pain and alteration in bowel habit). External physical examination confirmed the presence of external thoracolumbar fecal fistulae in association with a scoliosis. Abdominal examination was unremarkable. Diagnostic colonoscopy with biopsy did not definitively confirm a diagnosis of TB. Microscopic examination in both microbiology and histopathology labs failed to identify acid-fast bacilli; however, gastrointestinal TB (GITB) was considered high on the differential diagnosis list. It was elected to perform open laparotomy with resection of the left hemi-colon. Postoperatively, she was treated with standard anti-tuberculous treatment for 6 months. The patient had an uneventful postoperative course, during which the fistulae gradually closed over the next year. This case with several low-output fistulas exiting the patient's back was successfully treated by a combination of surgery and antimicrobial therapy. The aim of this rare case report is to raise the awareness of atypical presentation of tubercular colonic enterocutaneous fistula so that timely diagnosis and intervention can salvage patient quality of life.

Identification of miR-758-3p as Potential Modulator of CBX5 Expression in Gastric Cancer.

Gastric cancer is one of the most frequently diagnosed cancer types in China and also the leading causes of cancer-related death. Previous study showed chromobox 5 expression was elevated in gastric cancer, but little is known regarding the precise molecular mechanisms by which chromobox 5 expression was modulated. In this study, we revealed that chromobox 5 could promote gastric cancer cell proliferation, migration, and invasion in vitro. We screened and identified microRNA-758-3p, whose expression was downregulated in gastric cancer tissues and cell lines, which was a potential upstream molecule of chromobox 5. Upregulation of microRNA-758-3p could markedly downregulate the expression of chromobox 5. Additionally, expression of microRNA-758-3p and chromobox 5 was inversely correlated in gastric cancer tissues. Moreover, microRNA-758-3p overexpression suppressed gastric cancer cell proliferation, migration, and invasion, but these effects can be partially reversed by chromobox 5 overexpression. Collectively, our results indicate that microRNA-758-3p serves as a tumor suppressor and plays a crucial role in inhibiting the proliferation, migration, and invasion of gastric cancer via targeting chromobox 5 and implicate its potential application in cancer therapy.

ZNF217 is overexpressed and enhances cell migration and invasion in colorectal carcinoma.

BACKGROUND: To investigate the expression and clinical significance of zinc finger protein 217 (ZNF217) in human colorectal carcinoma (CRC). MATERIALS AND METHODS: The expression of ZNF217 in 60 CRC tissues and matched tumor adjacent tissues, collected between January 2013 and June 2014, was assessed immunohistochemically. The relationship between the expression of ZNF217 and clinicopathlogical features was analyzed by Pearson chi-square test. In addition, siRNA was used to down-regulate the expression of ZNF217 in CRC cells. The effects of ZNF217 for cell migration and invasion were measured by wound healing assay and transwell assay, respectively. RESULTS: The expression level of ZNF217 was significantly higher in CRC tissues than in tumor adjacent tissues (p<0.05), positively correlating with tumor size, lymphatic metastasis and advanced TNM stage (p<0.05). Down-regulation of ZNF217 in CRC cells could significantly suppress cell migration and invasion. CONCLUSIONS: ZNF217 is overexpressed in colorectal carcinoma tissues and is associated with tumor malignant clinicopathological features. ZNF217 may promote CRC progression by inducing cell migration and invasion.

Therapeutic options for intermediate-advanced hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is one of the most common malignancies, ranking the sixth in the world, with 55% of cases occurring in China. Usually, patients with HCC did not present until the late stage of the disease, thus limiting their therapeutic options. Although surgical resection is a potentially curative modality for HCC, most patients with intermediate-advanced HCC are not suitable candidates. The current therapeutic modalities for intermediate-advanced HCC include: (1) surgical procedures, such as radical resection, palliative resection, intraoperative radiofrequency ablation or cryosurgical ablation, intraoperative hepatic artery and portal vein chemotherapeutic pump placement, two-stage hepatectomy and liver transplantation; (2) interventional treatment, such as transcatheter arterial chemoembolization, portal vein embolization and image-guided locoregional therapies; and (3) molecularly targeted therapies. So far, how to choose the therapeutic modalities remains controversial. Surgeons are faced with the challenge of providing the most appropriate treatment for patients with intermediate-advanced HCC. This review focuses on the optional therapeutic modalities for intermediate-advanced HCC.