Construction of hiPSC-derived Excitatory Neural Network-like Tissue / 中山大学学报(医学科学版)

ObjectiveTo construct a neural network-like tissue with the potential of synaptic formation in vitro by seeding human induced pluripotent stem cell-derived neural precursor cells （hiPSC-NPCs） on decellularized optic nerve （DON）， so as to provide a promising approach for repair of nerve tissue injury. MethodsThrough directional induction and tissue engineering technology， human induced pluripotent stem cells （hiPSCs） and 3D DON scaffolds were combined to construct neural network-like tissues. Then the hiPSCs were directionally induced into human neural precursor cells （hNPCs） and neurons. Immunofluorescence staining was used to identify cell differentiation efficiency. 3D DON scaffolds were prepared. Morphology and cytocompatibility of scaffolds were identified by scanning electron microscopy and Tunnel staining. Induced hiPSC-NPCs were seeded on DON scaffolds. Immunofluorescence staining， scanning electron microscopy， transmission electron microscopy and patch clamp were used to observe the morphology and functional identification of constructed neural network tissues. Results①The results of immunofluorescence staining suggested that most of hiPSC-NPCs differentiated into neurons in vitro. We had successfully constructed a neural network dominated by neurons. ② The results of scanning electron microscopy and immunohistochemistry suggested that a neural network-like tissue with predominating excitatory neurons in vitro was successfully constructed. ③The results of immunohistochemical staining， transmission electron microscopy and patch clamp indicated that the neural network-like tissue had synaptic transmission function. ConclusionA neural network-like tissue mainly composed of excitatory neurons has been constructed by the combination of natural uniform-channel DON scaffold and hiPSC-NPCs， which has the function of synaptic transmission. This neural network plays a significant role in stem cell derived replacement therapy， and offers a promising prospect for repair of spinal cord injury （SCI） and other neural tissue injuries.

Construction of iPSC-derived Inhibitory Neural Network Tissue with Synaptic Transmission Potentials / 中山大学学报(医学科学版)

ObjectiveDirected differentiation of human induced pluripotent stem cells （hiPSCs） into spinal cord γ-aminobutyric acid （GABA）-ergic progenitor cells were implanted into an decellularized optical nerve （DON） bioscaffold to construct a hiPSC-derived inhibitory neural network tissue with synaptic activities. This study aimed to provide a novel stem cell-based tissue engineering product for the study and the repair of central nervous system injury. MethodsThe combination of stepwise directional induction and tissue engineering technology was applied in this study. After hiPSCs were directionally induced into human neural progenitor cells （hNPCs） in vitro， they were seeded into a DON for three-dimensional culture， allowing further differentiation into inhibitory GABAergic neurons under the specific neuronal induction environment. Transmission electron microscopy and whole cell patch clamp technique were used to detect whether the hiPSCs differentiated neurons could form synapse-like structures and whether these neurons had spontaneous inhibitory postsynaptic currents， respectively， in order to validate that the hiPSC-derived neurons would form neural networks with synaptic transmission potentials from a structural and functional perspective. ResultsThe inhibitory neurons of GABAergic phenotype were successfully induced from hiPSCs in vitro， and maintained good viability after 28 days of culture. With the transmission electron microscopy， it was observed that many cell junctions were formed between hiPSC-derived neural cells in the three-dimensional materials， some of which presented a synapse- like structure， manifested as the slight thickness of cell membrane and a small number of vesicles within one side of the cell junctions， the typical structure of a presynatic component， and focal thickness of the membrane of the other side of the cell junctions， a typical structure of a postsynaptic component. According to whole-cell patch-clamp recording， the hiPSC-derived neurons had the capability to generate action potentials and spontaneous inhibitory postsynaptic currents were recorded in this biotissue. ConclusionsThe results of this study indicated that hiPSCs can be induced to differentiate into GABAergic progenitor cells in vitro and can successfully construct iPSC-derived inhibitory neural network tissue with synaptic transmission after implanted into a DON for three-dimensional culture. This study would provide a novel neural network tissue for future research and treatment of central nervous system injury by stem cell tissue engineering technology.

Ginsenoside-Rg1 combined with a conditioned medium from induced neuron-like hUCMSCs alleviated the apoptosis in a cell model of ALS through regulating the NF-κB/Bcl-2 pathway / 中国天然药物

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease affecting both upper and lower motor neurons in the brain and spinal cord. One important aspect of ALS pathogenesis is superoxide dismutase 1 (SOD1) mutant-mediated mitochondrial toxicity, leading to apoptosis in neurons. This study aimed to evaluate the neural protective synergistic effects of ginsenosides Rg1 (G-Rg1) and conditioned medium (CM) on a mutational SOD1 cell model, and to explore the underlying mechanisms. We found that the contents of nerve growth factor, glial cell line-derived neurotrophic factor, and brain-derived neurotrophic factor significantly increased in CM after human umbilical cord mesenchymal stem cells (hUCMSCs) were exposed to neuron differentiation reagents for seven days. CM or G-Rg1 decreased the apoptotic rate of SOD1G93A-NSC34 cells to a certain extent, but their combination brought about the least apoptosis, compared with CM or G-Rg1 alone. Further research showed that the anti-apoptotic protein Bcl-2 was upregulated in all the treatment groups. Proteins associated with mitochondrial apoptotic pathways, such as Bax, caspase 9 (Cas-9), and cytochrome c (Cyt c), were downregulated. Furthermore, CM or G-Rg1 also inhibited the activation of the nuclear factor-kappa B (NF-κB) signaling pathway by reducing the phosphorylation of p65 and IκBα. CM/G-Rg1 or their combination also reduced the apoptotic rate induced by betulinic acid (BetA), an agonist of the NF-κB signaling pathway. In summary, the combination of CM and G-Rg1 effectively reduced the apoptosis of SOD1G93A-NSC34 cells through suppressing the NF-κB/Bcl-2 signaling pathway (Fig. 1 is a graphical representation of the abstract).

Changes of serum Golgi protein 73 expression and its clinical significance in breast cancer patients before and after radiotherapy / 中华内分泌外科杂志

Objective:To investigate the changes of serum Golgi protein 73 (GP73) expression and its clinical significance in breast cancer patients before and after radiotherapy.Methods:A total of 135 patients with primary breast cancer who were diagnosed and treated in Qingdao Hospital Affiliated to Shandong First Medical University Hospital from Aug. 2016 to Dec. 2017 were selected and received standard radiotherapy after surgery. Enzyme-linked immunosorbent assay (ELISA) was used to determine the content of GP73 in the serum of patients before and after radiotherapy, and the differences in the expression levels of GP73 before and after radiotherapy in patients with different molecular phenotypes, tumor stages, and pathological types were compared. Association between the expression of serum GP73 and clinical outcome before and after radiotherapy was analyzed.Results:The expression of GP73 in breast cancer patients was (117.69±33.57) mg/L before radiotherapy and (101.88±30.92) mg/L after radiotherapy, and the difference was statistically significant ( t=4.025, P<0.001) . Different molecular phenotypes were stratified and found that the expression of serum GP73 in patients with luminal A, luminal B, HER-2 overexpression, and triple negative after radiotherapy decreased compared with those before radiotherapy, but only the HER-2 overexpression type had statistically significant difference between before and after radiotherapy ( P<0.05) . Stratification according to different tumor stages showed that the expression of serum GP73 in patients with stage I, II, III and IV after radiotherapy was lower than that before radiotherapy, but only the patients with stage II, stage III, and stage IV were compared before and after radiotherapy, and the difference was statistically significant ( P<0.05) . Stratification according to different pathological types showed that the expression of serum GP73 in patients with invasive ductal carcinoma before and after radiotherapy was significantly lower in patients with invasive lobular carcinoma ( P<0.05) . In addition, the 1-year survival rate of the descending group was significantly higher than that of the ascending group, while the local recurrence rate and the occurrence of distant metastasis were significantly lower than those of the ascending group, and the difference was statistically significant ( P<0.05) . Conclusions:The expression level of serum GP73 in breast cancer patients after radiotherapy is significantly higher than that before radiotherapy, and it has a certain relationship with molecular phenotype, tumor stage, and pathological type. At the same time, the increase in serum GP73 expression after radiotherapy may be detrimental to the clinical outcome of patients.

The antitussive and expectorant mechanism of Platycodon total saponins based on metabonomics / 药学学报

UHPLC-Q-TOF/MS metabonomics technology was used to clarify the metabolic regulation pathways by which Platycodon total saponins (PTS) exert antitussive and expectorant effects in a mouse cough model, in which coughing is induced by concentrated ammonia, and in a phenol red excretion model. After approval by the Experimental Animal Ethics Committee of Jiangxi University of Chinese Medicine (Approval No. JZLLSC-20190235), the mice were randomly divided into a normal group, a model group, a positive drug group and a PTS group. Endogenous metabolites in mouse serum were identified by UHPLC-Q-TOF/MS. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used for multivariate analysis. Metabolic pathways were analyzed by the Metaboanalyst platform. The results show that PTS can significantly prolong the cough latent period and cough frequency of mice, and significantly increase phenol red excretion. UHPLC-Q-TOF/MS identified 19 metabolites related to cough, and PTS significantly decreased 16 of them; 17 metabolites related to expectoration were identified, and PTS decreased the levels of all. Metabolic pathway analysis showed that linoleic acid metabolism, arachidonic acid metabolism and glycerophospholipid metabolism were the main pathways involved in serum metabolite changes in this mouse cough model. Linoleic acid metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, arachidonic acid metabolism, phenylalanine metabolism and α-linolenic acid metabolism were the main pathways involved in serum metabolite changes in the phenol red excretion model. This study is the first to elucidate the regulation of antitussive and expectorant metabolic pathways and the effect of PTS on these pathways.

Kuanxiong Aerosol () in Treatment of Angina Pectoris: A Literature Review and Network Pharmacology / 中国结合医学杂志

Angina pectoris (AP) is the most common symptom of cardiovascular diseases, which seriously affects the quality of life in cardiovascular patients. Kuanxiong (KX) Aerosol (), a compound preparation that consists of 5 traditional Chinese medicines: Herba Asari , Rhizoma Alpiniae Officinarum, Lignum Santali Albi, Fructus Piperis Longi, and Borneolum, has been used in the treatment of AP for many years, exhibiting a significant curative effect and less side-effect. For the convenience and comprehensive understanding of KX Aerosol, this review systematically summarizes evidence on KX Aerosol in the treatment of AP including the pharmacological effects of its composition, clinical research, animal experiments, and network pharmacology prediction. Meanwhile, we highlight the research limitation of KX Aerosol at present. This review may guide the clinical application of KX Aerosol and further provide a reference for the research of AP.

Herbal textual research on Chuanxiong Rhizoma in Chinese classical prescriptions / 中国中药杂志

Fourteen classical prescriptions in the Catalog of 100 Ancient Classical Prescriptions(First Batch) promulgated in 2018 contain Chuanxiong Rhizoma, which reveals the high medicinal value and wide application of Chuanxiong Rhizoma. This paper systematically reviews the ancient herbal books and modern literature to explore the name, origin, genuine producing area, medicinal part, harvesting, and processing of Chuanxiong Rhizoma, thus facilitating the development of classical prescriptions containing Chuan-xiong Rhizoma. It is confirmed that Chuanxiong Rhizoma, formerly known as "Xiongqiong" in Chinese, was first called "Chuanxiong" in late Tang Dynasty, which has been gradually accepted as its official name due to the rise of the status of Chuanxiong Rhizoma produced in Sichuan. The main original plant of Chuanxiong Rhizoma in past dynasties has always been deemed to be Ligusticum chuan-xiong(Umbellifera), whose rhizome serves as the medicinal part. In general, it is best harvested in summer but the harvesting time can vary with different growth environments. Since the Song Dynasty, Sichuan province has been recognized as the genuine producing area of Chuanxiong Rhizoma in light of the high yield and good quality. It is suggested that Chuanxiong Rhizoma from Sichuan be used preferentially in the development of classical prescriptions. There are multiple processing methods of Chuanxiong Rhizoma recorded in ancient medical classics, and the raw(after purifying and slicing) or wine-processed or stir-fried Chuanxiong Rhizoma is still in use today. In the development of classical prescriptions containing Chuanxiong Rhizoma, Chuanxiong Rhizoma is advised to be processed in accordance with current processing standards if the specific processing method is described in the medical classics. If not, the raw Chuanxiong Rhizoma is preferred and then processed following the processing standards of Chuanxiong Rhizoma decoction pieces in Chinese Pharmacopoeia.

Effect of indwelling drainage tube and extubation time on recessive hemorrhage and functional recovery after total hip arthroplasty / 中国骨伤

OBJECTIVE@#To study and compare the effect of indwelling drainage tube and extubation time on occult hemorrhage and functional recovery after total hip arthroplasty(THA).@*METHODS@#From July 2017 to June 2018, 123 patients who underwent THA in our hospital for the first time were selected as the subjects of study. According to whether the drainage tube was retained or not and the time of extubation, they were divided into three groups:in group A, 41 patients (24 males, 17 females, age 53 to 77 years) did not put drainage tube after THA;in group B, 41 patients were removed 24 hours after THA, 26 males and 15 females, aged 55 to 74 years;in group C, 41 patients were removed 48 hours after THA, 25 males and 16 females, aged 52 to 75 years. The VAS score of pain 72 hours after THA, the total and recessive blood loss, the time of starting functional exercise, and the incidence of postoperative limb swelling were recorded. All the patients were followed up for one year after discharge. Harris hip score was used to evaluate the degree of hip function recovery one year after operation.@*RESULTS@#The occult blood loss of group A, B and C were(513.6±25.3), (521.7±33.4), (519.3±29.8) ml, respectively, with no significant difference(>0.05). There was no significant difference in blood loss in operation among the three groups(>0.05). In group B and C, the postoperative apparent blood loss was more than that in group A (0.05). The time of getting out of bed in group A was shorter than that in group B and C (0.05). There was no significant difference in the incidence of complications among three groups (>0.05).@*CONCLUSION@#Whether the drainage tube is left or not and the time of extubation have no significant effect on the latent blood loss and functional recovery after THA, but without drainage tube after THA can reduce the apparent blood loss, patients can get out of bed at 6 hours after THA, which is more conducive to the recovery and nursing of patients.

Screening of Components with Potential Regulation Effect in Lagotis brevituba Based on Target Cell and Molecular Docking / 中国实验方剂学杂志

Objective:To study the components with urate anion transporter 1（URAT1） regulation effect and their combination mechanisms of Lagotis brevituba by integrating techniques of HK-2 cell capture，UPLC-Q-TOF-MS and molecular docking，so as to provide material and theory bases for the development of new hypouricemic medicines based on L. brevituba. Method:The HK-2 cells were applied to capture the components of L. brevituba. UPLC-Q-TOF-MS was used to identify those components. The molecular docking technique was adopted to study the interaction mechanism between the compounds and URAT1. Result:Eight components were successfully screened and identified as hyperoside，plantamajoside，kaempferol-3-O-glucoside，lugrandoside，nepitrin，isolugrandoside，homoplantaginin，luteolin，respectively. Those components could combine with URAT1 mainly through hydrogen bond，van der Waals force and hydrophobic action，which were closely related to structure and compound types. Furthermore，the LibDock score of phenylethanoids was higher than that of flavonoids. Conclusion:The integration of target cell capture，UPLC-Q-TOF-MS and molecular docking techniques could be successfully used to identify captured compounds of L. brevituba with URAT1 regulation effects and illustrate their potential combination mechanisms as well as the structure-activity relationships. The findings may provide material and theory bases for the development of new hypouricemic medicines based on L. brevituba.

Follow-up of NSCLC patients with first-generation EGFR-TKI resistance using third-generation EGFR-TKI / 中国医师杂志

Objective:To analyze the clinical efficacy of epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) in patients with advanced NSCLC.Methods:We collected 23 cases of NSCLC advanced patients, who were treated in the affiliated Cancer Hospital of Guangzhou Medical University from January 2015 to March 2020. And these 23 cases of patients with first-generation EGFR-TKIs resistance were treated with the third-generation EGFR-TKI drugs. We analyzed their clinicopathological characteristics, studied their therapeutic effects, and followed up their progression-free survival (PFS).Results:It is showed that 16 of 23 cases (69.56%) were got local progression and 7 of 23 cases (31.43%) were found with systemic progression. Briefly, the median PFS of the 23 patients was 17.5 months. A total of 7 cases occurred rashes after taking EGFR-TKI, and 3 cases got abnormal liver function. Fortunately, they were all improved after symptomatic treatments. Additionally, no bone marrow suppression (granulocytes, neutropenia, thrombocytopenia, anemia) and digestive tract reactions (nausea, vomiting, diarrhea) were occurred in 23 cases of NSCLC patients. The mental and physical improvement of EGFR-TKI in the third generation of 19 patients was more obvious than that in the first generation of EGFR-TKI. Among them, 15 cases showed more obvious lesion shrinkage after third-generation EGFR-TKI treatment. 4 patients with GGO had cleaner disappearance than that of the first-generation EGFR-TKI.Conclusions:Compared with traditional chemotherapy, the first-generation EGFR-TKI resistance treatment with three-generation EGFR-TKI treatment has better efficacy with reduced toxic and side effects, and significantly improved the life quality of advanced NSCLC patients.

Catheter balloon dilation combined with acupuncture for cricopharyngeal achalasia after brain stem infarction: a randomized controlled trial / 中国针灸

OBJECTIVE@#To screen the optimal acupuncture regimen for cricopharyngeal achalasia (CPA) after brain stem infarction and compare the therapeutic effect between the combined therapy of catheter balloon dilation and acupuncture and the simple application of catheter balloon dilation.@*METHODS@#The patients suffering from neuropathic dysphagia in CPA after brain stem infarction were selected as the subjects. After confirmed in the diagnosis with video fluoroscopic swallowing study (VFSS), they were randomized into 6 groups, 15 cases in each one, named group A (routine treatment), group B (catheter balloon dilation), group C1 (treated with acupuncture in local area), group C2 (treated with acupuncture based on differentiation), group C3 (treated with acupuncture at the local area and the acupoints based on differentiation) and group D (catheter balloon dilation combined with the optimal acupuncture). Two phases were included in the study. In the first phase of study, the therapeutic effect was compared among the three acupuncture groups, named C1, C2 and C3 group, so as to screen the optimal acupuncture regimen. In the group C1, the main acupoints included Fengchi (GB 20), Wangu (GB 12), Yifeng (TE 17) and three-tongue points (Extra). In the group C2, the main acupoints were Neiguan (PC 6), Tongli (HT 5), Zusanli (ST 36) and Sanyinjiao (SP 6) as well as the supplementary acupoints in accordance with the syndrome differentiation. In the group C1 and group C2, after , the electroacupuncture was used, with continuous dense wave, 5 to 8 Hz in frequency. The needles were retained for 30 min. Acupuncture was given once a day, 5 treatments a week. Before treatment, in 6 weeks of treatment or after removal of gastric tube, the rehabilitation was evaluated. In the group C3, the acupoints, manipulation and treating course were same as the group C1 and group C2. In the 2nd phase of study, theresults of rehabilitation treatment were compared among the group A, group B and group D. The treatment was given once a day, 5 times a week. Before treatment, after gastric tube removal or in 6 weeks of treatment, the evaluation was conducted. The feedingswallowing function grade and VFSS were adopted in the evaluation among the above 6 groups.@*RESULTS@#① In the VFSS comparison at 1st phase of study after treatment, the food transporting ability at oral dysphagia, the results in the group C3 and group C1 were better obviously than the group C2 (both 0.05). The severity of dysphagia in the group C3 was milder than the group C2 and group A (both <0.05). ② In the VFSS comparison at the 2nd phase of study, for the food transporting ability, the results in the group D and the group B were obviously better than the group A (both <0.05). Regarding the function at the pharyngeal dysphagia and aspiration, the results in the group D were better than the group B and group A, those in the group B were better than the group A (all <0.05). The difference in the extubation rate among the group A, group B and group D after treatment was significant statistically (<0.01), of which, the extubation rate in the group D was the highest and the rate in the group A was the lowest. The dysphagia degree in the group D was milder than the group B and group A and that in the group B was milder than the group A (all <0.05).@*CONCLUSION@#In the study of the different acupuncture methods, the acupuncture at the local acupoints and the acupoints selected based on differentiation is the optimal acupuncture regimen for cricopharyngeal achalasia after brain stem infarction. The catheter balloon dilation combined with acupuncture present the synergistc effect on cricopharyngeal achalasia after brain stem infarction, obviously relieve dysphagia and reduce aspiration.

Methodology for Screening Effective Substances of Chinese Herbal Medicine for Reducing Serum Uric Acid Level / 中国实验方剂学杂志

Gout is caused by the nucleation and growth of monosodium rate crystals in tissues and around joints, which is followed by long-standing hyperuricemia and serum urate of above the saturation threshold. It could cause a series of complications, such as cardiovascular, hypertension, and renal complications. Over the past two decades, the incidences of hyperuricemia and gout have been increasing due to the continuous improvement of living standards and the changes in dietary structure. The prime and most important therapy for hyperuricemia and gout is to reduce serum uric acid levels, but the western medicine for reducing uric acid in clinical application has serious toxic and side effects. With the rapid development of modern science and technology, the application and development of different screening methods for effective ingredients with a low toxicity and side effects from Chinese herbal medicines for reducing serum uric acid levels has attracted much attention in the research and development of drugs for the prevention and treatment of hyperuricemia and gout. In this study, the screening methods for extracts, fractions, active monomer components and other effective substances were reviewed and analyzed. According to the findings, the screening methods had a considerable progress both in vivo and in vitro. The results showed that the in vivo methods were mainly applied for studying the urate lowing effect and mechanisms of herbal extracts, while the studies for xanthine oxidase(XOD) inhibitors mainly depended on the in vitro methods. Molecular docking homology modeling and liquid chromatography-mass spectrometry have become a new trend for screening effective substances with XOD inhibitory activities and uric acid excretion activities, while cell model will open up a new way for screening effective substances for uric acid excretion. The review provides certain reference for effective components screening of hyperuricemia and gout.

Research advances on the role of extracellular matrix in pulmonary fibrosis / 基础医学与临床

Pulmonary fibrosis(PF)is characterized by extensive deposition of extracellular matrix(ECM),changes in biomechanical properties of ECM result from the process of PF actively drive disease progression.Several matrix protein candidates correlate with fibrotic pathologies,ECM may offer many novel therapeutic targets.

Effect of puerarin on inflammation-related diseases / 中国药理学通报

Puerarin is a kind of isoflavone extracted from the root of Radix Pueraria .Studies showed that puerarin had a variety of functional activities, such as anti-inflammation, antioxidation, anti-osteoporosis , lowering blood glucose , anti-tumor and so on . The effects of puerarin for inflammation-related diseases are mainly summarized in this research , including cardiovascular disease, osteoporosis, diabetes and cancer .Realizing that anti-inflammation is the common mechanism for puerarin in treatment of these diseases , reference basis is expected to be offered for puerarin to play a broad application prospect in clinical practice .

Clinical value of superior mesenteric vein (No.14v) lymph node dissection in D2 gastrectomy for locally advanced distal gastric cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore the value of superior mesenteric vein (No.14v) lymph node dissection in D2 gastrectomy for locally advanced distal gastric cancer.</p><p><b>METHODS</b>A retrospective cohort study was carried out. A total of 796 patients with locally advanced distal gastric cancer undergoing D2 gastrectomy at the Cancer Center of Guangzhou Medical University between 2002 and 2016 were enrolled.</p><p><b>INCLUSION CRITERIA</b>locally advanced distal gastric adenocarcinoma confirmed by postoperative pathology; adenocarcinoma located at or invaded into lower 1/3 stomach; lymphadenectomy was D2 or D2+; negative resection margin confirmed by pathology; no distal metastasis was found; preoperative neoadjuvant chemotherapy was not administrated. Patients with undefined group of lymph nodes by postoperative pathology and those who were died perioperatively were excluded. Among 796 patients, 293 underwent No.14v dissection (No.14vD+ group) and the other 503 patients did not undergo No.14v dissection (No.14vD- group). The 5-year overall survival was compared between the two groups. Therapeutic index of No.14v lymph nodes was calculated according to the following formula: therapeutic index=metastatic rate of No.14 lymph nodes (%) × 5-year survival rate of patients with No.14 lymph node metastasis(%) × 100. Meanwhile, stratified analyses based on pathological TNM staging were performed.</p><p><b>RESULTS</b>There were no significant differences in age, gender, tumor size, Borrmann type, Lauren classification, histological type, surgical procedure, and number of harvested lymph node between two groups (all P>0.05). However, compared to No.14vD- group, No.14vD+ group had more advanced T staging (χ² =14.771, P=0.005) and TNM staging (χ² =18.339, P=0.003), and higher ratio of receiving adjuvant chemotherapy (χ² =4.205, P=0.040). The median follow-up period was 47 months. The 5-year survival rate in No.14vD+ and No.14vD- groups was 57.4% and 46.8% respectively without statistically significant difference (P=0.313). After adjusting for confounding factors, Cox proportional hazards model showed that No.14v lymphadenectomy was not an independent prognostic factor(HR=0.802, 95%CI: 0.545-1.186, P=0.124). Stratified analyses revealed that in all TNM stages, 5-year survival rates were not significantly different between two groups (all P>0.05). However, No.14v lymphadenectomy showed a tendency of survival benefit when the tumor staging after advancing to III A stage(III A: P=0.103; III B: P=0.085; III C: P=0.060). Five-year survival rates of No.14vD+ and No.14vD- groups in stage III A were 54.9% and 45.2%, in III B stage were 39.8% and 29.5%, in III C stage were 27.5% and 16.2%, respectively. After combining III A, III B and III C, the No.14vD+ group had a higher 5-year survival rate than No.14vD- group (39.2% vs. 27.7%, P=0.006). The No.14v metastasis rate in No14v+ group was 12.6%(37/293), including 0%(0/46), 2.5%(1/40), 4.9%(2/41), 15.7%(8/51), 20.8%(11/53) and 24.2%(15/62) in stages I B, II A, II B, III A, III B and III C respectively. The metastasis rate of No.14v lymph node in stage III patients was 20.5%(34/166). The 5-year survival rate of these 34 stage III patients with No.14v metastasis was 21.1%. The therapeutic index of No.14v lymph node in stage III patients was 4.3, which was comparable with 3.9 of No.9 and 4.9 of No.11p, even higher than 2.6 of No.1.</p><p><b>CONCLUSIONS</b>Although No.14v lymphadenectomy can not improve the overall survival of patients with locally advanced distal gastric cancer, but it may significantly improve survival in those with stage III cancer. The therapeutic index of No.14v lymph node is similar to No.2 station lymph node in patients with stage III distal gastric cancer. Therefore No.14v lymph node should be included in D2 dissection.</p>

Research on XOD captured components in Lagotis brevituba based on UPLC-Q-TOF-MS and molecular docking technology / 中国中药杂志

Potential xanthine oxidase (XOD) inhibitors in Lagotis brevituba were captured by using affinity and ultrafiltration. The structures of the captured components were identified by ultra-performance liquid chromatography coupled with Q-TOF mass spectrometry (UPLC-Q-TOF-MS). The binding intensity and binding mechanism between the captured components and XOD were analyzed by using molecular docking software Autodock 4.2. A total of 17 compounds were identified, including 9 flavonoids, 5 phenolic acids and 3 triterpenes. Molecular docking results showed that all the captured components could be spontaneously bound with XOD mainly via hydrogen bond, Van der Waals' force and hydrophobic interaction. From the perspective of binding energy and scoring function, the collected fractions all had potential prospects for XOD inhibitors, and the flavonoid luteolin-3',7 glucuronide had the best effect. The results also showed that affinity and ultrafiltration, ultra-performance liquid chromatography-mass spectrometry (UPLC-MS) and molecular docking technology can provide a powerful tool for the analysis of XOD inhibitor components in natural products.

Determination of Seven Urinary Metabolites of Organophosphate Esters Using Liquid Chromatography-Tandem Mass Spectrometry / 分析化学

A simple method was developed for simultaneous determination of seven urinary metabolites of organophosphate esters by liquid chromatography-tandem mass spectrometry (LC-MS / MS). Based on different physical and chemical properties of these OPs metabolites, the solid phase extraction cartridges and the washing and eluting solvents were optimized in details. Furthermore, the mobile phase and mass spectrometric parameters were also investigated. The results showed that Oasis WAX cartridge was the best SPE column in this study, and 2 mL of NH3 ·H2 O (5% ) in methanol and 2 mL of methanol were chosen as the eluting solvents. The recoveries of six analytes were ranged from 60. 5% to 104. 0% , whereas DEP ranged from 17. 8% to 36. 2% . Seven analytes could be baseline separated from each other under the optimized chromatographic conditions. The limits of detection and quantification of seven analytes ranged from 0. 005 to 0. 2 μg / L and 0. 02 to 0. 5 μg / L, respectively. The standard deviations of response repeatability for intra-day and inter-day period were lower than 15. 4% . This method was finally applied to determination of metabolites of OPs from 10 urines from general population in Guangzhou city. The concentrations of total OPs metabolites in urine samples ranged from 0. 5 to 6. 7 μg / L.

Fast identification of constituents of Lagotis brevituba by using UPLC-Q-TOF-MS/MS method / 中国中药杂志

The chemical constituents of Lagotis brevituba were rapidly determined and analyzed by using ultra performance liquid chromatography tandem quadrupole time of flight mass spectrometry (UPLC-Q-TOF-MS/MS) method, providing material basis for the clinical application of L. brevituba. The separation was performed on UPLC YMC-Triart C₁₈ (2.1 mm×100 mm, 1.9 μm) column, with acetonitrile-water containing 0.2% formic acid as mobile phase for gradient elution. The flow rate was 0.4 mL•min-1 gradient elution and column temperature was 40 ℃, the injection volume was 2 μL. ESI ion source was used to ensure the data collected in a negative ion mode. The chemical components of L. brevituba were identified through retention time, exact relative molecular mass, cleavage fragments of MS/MS and reported data. The results showed that a total of 22 compounds were identified, including 11 flavones, 6 phenylethanoid glycosides, 1 iridoid glucosides, and 4 organic acid. The UPLC-Q-TOF-MS/MS method could fast identify the chemical components of L. brevituba, providing valuable information about L. brevituba for its clinical application.

Prophylactic administration of all-trans retinoic acid alleviates inflammation in rats with collagen-induced arthritis / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effects of prophylactic administration of all-trans retinoic acid (ATRA) in relieving inflammation in a rat model of collagen-induced arthritis (CIA).</p><p><b>METHODS</b>Female Wistar rats (6 to 8 weeks old) were randomly divided into normal control group, solvent control group, and prophylactic ATRA treatment (0.05, 0.5, and 5 mg/kg) groups. All the rats except for those in normal control group were subjected to subcutaneous injection of type II collagen and incomplete Freund adjuvant in the tails to induce CIA, followed by injection on the following day with saline, corn oil or different doses of ATRA 3 times a week. The arthritis index (AI) scores, histological scores, serum levels of TNF-α, IL-17A, and IL-10, and expressions of proteases related with cartilage damage were evaluated.</p><p><b>RESULTS</b>On the 15th day after the primary immunization, the AI scores increased significantly in all but the normal control groups; the scores increased progressively in all the 3 ATRA groups but remained lower than that in the solvent control group, which was stable over time. The rats in the 3 ATRA groups showed obvious pathologies in the knee and ankle joints, but the semi-quantitative scores of pathology damage showed no significance among them. Compared with those in solvent control group, the serum IL-17A and TNF-α levels decreased, serum IL-10 level increased, and the expressions of ADAMT-4 and MMP-3 proteins decreased significantly in the knees in the 3 ATRA groups.</p><p><b>CONCLUSION</b>ATRA can reduce the production of TNF-α and IL-17A and increase the production of IL-10 to alleviate the inflammation in rats with CIA. ATRA may delay the progression of RA by correcting the imbalance of Th1/Th2 and Th17/Treg.</p>

Study on genetic polymorphism of Platycodon grandiflorum based on barcoding of ITS2 / 中国中药杂志

<p><b>OBJECTIVE</b>ITS2 of DNA barcoding was used to study genetic polymorphism of Platycodon grandiflorum.</p><p><b>METHOD</b>Total genomic DNA was isolated from P. grandiflorum. PCR was used to amplified the region of internal transcribed spacer 2 (ITS2), and PCR products were sequenced. The sequences of ITS2 were analyzed and compared by Clustal. The intraspecies genetic distance was calculated based on Kimura 2-parameter model by using MEGA 5.05. The ITS2 sequence of Codonopsis pilosula was used as the outreach value for plants of the genus, and the phylogenic tree used constructed by Neighbor-Joining (NJ) method.</p><p><b>RESULT</b>The K2-P's genetic distance of all samples were ranged from 0 to 0.930. The K2-P's genetic distance of samples at the same area were ranged from 0 to 0.178. The K2-P's genetic distance of samples at different areas were ranged from 0.735 to 0.930. The analytical result showed that the degree of genetic variation were heavy in intraspecies of P. grandiflorum and significantly correlated with geographical location.</p><p><b>CONCLUSION</b>The DNA barcoding of ITS2 can applied to study the intraspecific genetic diversity, it provides a reference for further development of DNA barcoding technology applications.</p>