Comparison of Conduits Fabricated by Fresh and Predegenerated Skeletal Muscles for Peripheral Nerve Repairing.

ABSTRACT: Reconstruction of peripheral nerve injury remains a challenge for clinical medicine. Previous reports have confirmed that external oblique muscle-fabricated nerve conduit (EMC) could effectively be used to promote peripheral nerve regeneration. In this study, we compared between conduits fabricated from fresh muscle and conduits fabricated from predegenerated muscle for the repair of peripheral nerve defects in a mouse sciatic nerve transection model. We found that the number, diameter, and myelin sheath thickness of the myelinated nerve fibers of the regenerative nerve in the EMC group were larger than those of the predegenerated-EMC (P-EMC) group eight weeks after surgery. The sciatic function index and gastrocnemius wet-weight mass ratio in the EMC group were higher than those in the P-EMC group. The Bcl-2/Bax ratio and the number of Schwann cell nucleus in the proximal nerve stumps in the EMC group were greater than those in the P-EMC group. In conclusion, our results confirmed that the use of fresh skeletal muscle nerve conduit increased the Bcl-2/Bax ratio and promoted the survival of Schwann cells of the proximal nerve stump compared with that of predegenerated skeletal muscle nerve conduits, thus achieving better functional recovery after sciatic nerve defect.

[Overview of research and development of polypeptide drugs and traditional Chinese medicine-peptides].

Peptide is a compound consisting of 2-50 amino acids, which is intermediate between small molecule and protein. It is characterized by a variety of biological activities, easy absorption, strong specific targeting, and few side effects and has become one of the hotspots in biomedical research in recent years. Chinese medicine contains a large number of peptides. The traditional processing methods such as decocting and boiling can effectively boost peptides to exert their due biological activities. At present, however, the research on Chinese medicinal components in laboratory generally employs high-concentration alcohol extraction method, which may cause the peptides to be ignored in many natural Chinese medicines. Substantial studies have revealed that the peptides in Chinese medicine are important material basis responsible for the traditional efficacy. Based on years of research and literature retrieval, this study put forward the concept of "traditional Chinese medicine(TCM)-peptides", referring to the components consisting of two or more amino acids with molecular weight between small molecules and proteins that can express the efficacy of Chinese medicine. Furthermore, this study also summarized the extraction and separation of TCM-peptides, and structure determination methods and routes, predicted the research prospect of modern research methods of TCM-peptides based on "holistic view" and big data. The artificial intelligence prediction was combined with high-throughput screening technology to improve the discovery efficiency and accuracy of TCM-peptides, and holographic images between TCM-peptides and biological targets were established to provide references for the innovative drug design and related health product development of TCM-peptides based on TCM theories.

Metabolic Activation of Elemicin Leads to the Inhibition of Stearoyl-CoA Desaturase 1.

Elemicin is a constituent of natural aromatic phenylpropanoids present in many herbs and spices. However, its potential to cause toxicity remains unclear. To examine the potential toxicity and associated mechanism, elemicin was administered to mice for 3 weeks and serum metabolites were examined. Enlarged livers were observed in elemicin-treated mice, which were accompanied by lower ratios of unsaturated- and saturated-lysophosphatidylcholines in plasma, and inhibition of stearoyl-CoA desaturase 1 (Scd1) mRNA expression in liver. Administration of the unsaturated fatty acid oleic acid reduced the toxicity of 1'-hydroxylelemicin, the primary oxidative metabolite of elemicin, while treatment with the SCD1 inhibitor A939572 potentiated its toxicity. Furthermore, the in vitro use of recombinant human CYPs and chemical inhibition of CYPs in human liver microsomes revealed that CYP1A1 and CYP1A2 were the primary CYPs responsible for elemicin bioactivation. Notably, the CYP1A2 inhibitor α-naphthoflavone could attenuate the susceptibility of mice to elemicin-induced hepatomegaly. This study revealed that metabolic activation of elemicin leads to SCD1 inhibition in liver, suggesting that upregulation of SCD1 may serve as potential intervention strategy for elemicin-induced toxicity.

A metabolomic perspective of pazopanib-induced acute hepatotoxicity in mice.

To elucidate the metabolism of pazopanib, a metabolomics approach was performed based on ultra-performance liquid chromatography coupled with electrospray ionization quadrupole mass spectrometry. A total of 22 pazopanib metabolites were identified in vitro and in vivo. Among these metabolites, 17 were novel, including several cysteine adducts and aldehyde derivatives. By screening using recombinant CYPs, CYP3A4 and CYP1A2 were found to be the main forms involved in the pazopanib hydroxylation. Formation of a cysteine conjugate (M3), an aldehyde derivative (M15) and two N-oxide metabolites (M18 and M20) from pazopanib could induce the oxidative stress that may be responsible in part for pazopanib-induced hepatotoxicity. Morphological observation of the liver suggested that pazopanib (300 mg/kg) could cause liver injury. The aspartate transaminase and alanine aminotransferase in serum significantly increased after pazopanib (150, 300 mg/kg) treatment; this liver injury could be partially reversed by the broad-spectrum CYP inhibitor 1-aminobenzotriazole (ABT). Metabolomics analysis revealed that pazopanib could significantly change the levels of L-carnitine, proline and lysophosphatidylcholine 18:1 in liver. Additionally, drug metabolism-related gene expression analysis revealed that hepatic Cyp2d22 and Abcb1a (P-gp) mRNAs were significantly lowered by pazopanib treatment. In conclusion, this study provides a global view of pazopanib metabolism and clues to its influence on hepatic function.

PPARα Mediates the Hepatoprotective Effects of Nutmeg.

Nutmeg is a Traditional Chinese Medicine used to treat gastrointestinal diseases. Some reports have indicated that nutmeg has hepatoprotective activity. In this study, a thioacetamide (TAA)-induced acute liver injury model in mice was used to explore the mechanism of the protective effects of nutmeg extract (NME), including its major bioactive component myrislignan. The results indicated that NME could effectively protect TAA-induced liver damage as assessed by recovery of increased serumtransaminases, decrease in hepatic oxidative stress, and lower hepatic inflammation. Metabolomics analysis further revealed that treatment with NME led to the recovery of a series of lipids including lysophosphatidylcholines that were decreased and a lowering of acylcarnitines that were increased in mouse plasma and liver after TAA exposure. Gene expression analysis demonstrated that the hepatoprotective effect of NME was achieved by modulation of the peroxisome proliferator-activated receptor alpha (PPARα) as well as the decrease in oxidative stress. NME could not protect from TAA-induced liver injury in Ppara-null mice, suggesting that its protective effect was dependent on PPARα. Myrislignan, a representative neolignan in nutmeg, showed potent protective activity against TAA-induced liver toxicity. These data demonstrate that nutmeg alleviates TAA-induced liver injury through the modulation of PPARα and that the lignan compounds in nutmeg such as myrislignan partly contributed to this action.

Validation of the moderate severity category of acute pancreatitis defined by determinant-based classification.

BACKGROUND: Recent international multidisciplinary consultation proposed the use of local (sterile or infected pancreatic necrosis) and/or systemic determinants (organ failure) in the stratification of acute pancreatitis. The present study was to validate the moderate severity category by international multidisciplinary consultation definitions. METHODS: Ninety-two consecutive patients with severe acute pancreatitis (according to the 1992 Atlanta classification) were classified into (i) moderate acute pancreatitis group with the presence of sterile (peri-) pancreatic necrosis and/or transient organ failure; and (ii) severe/critical acute pancreatitis group with the presence of sterile or infected pancreatic necrosis and/or persistent organ failure. Demographic and clinical outcomes were compared between the two groups. RESULTS: Compared with the severe/critical group (n=59), the moderate group (n=33) had lower clinical and computerized tomographic scores (both P<0.05). They also had a lower incidence of pancreatic necrosis (45.5% vs 71.2%, P=0.015), infection (9.1% vs 37.3%, P=0.004), ICU admission (0% vs 27.1%, P=0.001), and shorter hospital stay (15+/-5 vs 27+/-12 days; P<0.001). A subgroup analysis showed that the moderate group also had significantly lower ICU admission rates, shorter hospital stay and lower rate of infection compared with the severe group (n=51). No patients died in the moderate group but 7 patients died in the severe/critical group (4 for severe group). CONCLUSIONS: Our data suggest that the definition of moderate acute pancreatitis, as suggested by the international multidisciplinary consultation as sterile (peri-) pancreatic necrosis and/or transient organ failure, is an accurate category of acute pancreatitis.

Fatal multiple acyl-CoA dehydrogenase deficiency caused by ETFDH gene mutation: A case report.

BACKGROUND: Multiple acyl-CoA dehydrogenase deficiency (MADD) is a disease of rare autosomal recessive disorder. There are three types of MADD. Type I is a neonatal-onset form with congenital anomalies. Type II is a neonatal-onset form without congenital anomalies. Type III is considered to a milder form and usually responds to riboflavin. However, late-onset form could also be fatal and not responsive to treatments. CASE SUMMARY: We report a severe case of a young man with onset type III MADD induced by drugs and strenuous exercise characterized by rhabdomyolysis and liver dysfunction. Urine analysis indicated 12 out of 70 kinds of organic acids like glutaric acid-2 were detected. Serum analysis in genetic metabolic diseases revealed 24 out of 43 tested items were abnormal, revealing the elevation of several acylcarnitines and the reduction of carnitine in the patient. By next generation sequencing technology for gene sequencing related to fatty acid oxidation and carnitine cycle defects, a rare ETFDH gene variant was identified: NM_004453:4:C.1448C>T(p.Pro483 Leu). The patient was diagnosed with late-onset GAII. He was not responsive to riboflavin and progressively worsened into multiple organ failure that finally led to death. CONCLUSION: Type III MADD can also be fatal and not responsive to treatments.

Harmonizing international quality standards for Andrographis paniculata: A comparative analysis of content determination methods across pharmacopeias.

The quality standards for Andrographis paniculata, a widely used medicinal herb, exhibited significant variations across different pharmacopeias. In this study, we compared the HPLC content determination methods and total lactone content of A. paniculata samples from different regions, as specified in the Chinese (CP), United States (USP), European (EP), Thai (TP), and Indian pharmacopeias (IP), as well as the Hong Kong Chinese Materia Medica Standards (HK). We aimed to assess the differences and similarities among these pharmacopeias and harmonized international quality standards for A. paniculata. The analysis revealed variations in sample preparation, liquid chromatographic conditions, fingerprint profiles, and total lactone content among the different pharmacopeias. Specifically, the CP and HK methods exhibited superior sample preparation and chromatographic separation. Further comparing the content of 20 A. paniculata samples with the CP, USP, EP and HK methods showed consistent determinations for the same components, indicating similar detection capabilities. The discrepancies in total lactone content primarily stemmed from differences in the number and types of detected compounds. Moreover, the acceptance criteria exhibited a stringency in the order CP > HK > EP > USP. In conclusion, this comparison analysis of content determination in CP, USP, HK, EP, TP and IP provided a scientific foundation for the international standardization and trade regulations of A. paniculata. It also served as a valuable reference for the development of international quality standards for other medicinal herbs, facilitating the harmonization of global pharmaceutical standards.

Human leukocyte antigen-DR expression on peripheral monocytes may be an early marker for secondary infection in severe acute pancreatitis.

BACKGROUND/AIMS: To investigate whether the human leukocyte antigen-DR (HLA-DR) expression on peripheral monocytes can be utilized as a precursor to a secondary infection of severe acute pancreatitis (SAP). METHODOLOGY: Patients diagnosed with SAP who were admitted into West China Hospital within 48 h after symptom onset from July 1, 2010 to December 31, 2010 (n = 40) were included. HLD-DR expression on peripheral monocytes on the 1st, 3rd, 5th and 7th day of hospitalization was detected with flow cytometry analysis to determine whether a prediction could be made in regards to development of a secondary infection. RESULTS: There were 11 patients with secondary infection complications, 4 of which died during hospitalization. On the 1st, 3rd, 5th and 7th day, HLA-DR expression on monocytes in the infected patients was lower than those in the noninfected patients (P < 0.05). There was no statistical significance in the serum CRP and APACHE II between the groups on the first day (P > 0.05). Upon initial admission HLA-DR expression showed a negative correlation with longer-term admission APACHE II (r = -0.790, P = 0.000) and serum CRP (r = -0.642, P = 0.000). The area under the ROC curve (AUC) was 0.837 (95%CI: 0.685-0.989, P = 0.001) for admission HLA-DR, 0.809 (95% CI: 0.667-0.951; P = 0.003) for APACHE II score and 0.781 for serum CRP (95% CI: 0.627-0.934; P = 0.007) to predict secondary infection. The cut-off value of prediction of secondary infection was 35.8% in HLA-DR expression with a sensitivity of 81.8% and a specificity of 82.8%, 10.5 in APACHE II on admission with a sensitivity of 90.9% and a specificity of 48.3%, 155 mg/L in serum CRP on admission with a sensitivity of 90.9% and a specificity of 44.8%. CONCLUSIONS: The HLA-DR expression on monocytes may be an ideal marker for an early prediction of secondary infection in SAP.

Urinary trypsinogen-2 for diagnosing acute pancreatitis: a meta-analysis.

BACKGROUND: Currently, serum amylase and lipase are the most popular laboratory markers for early diagnosis of acute pancreatitis with reasonable sensitivity and specificity. Urinary trypsinogen-2 (UT-2) has been increasingly used but its clinical value for the diagnosis of acute pancreatitis and post-endoscopic retrograde cholangiopancreatography (ERCP) pancreatitis has not yet been systematically assessed. DATA SOURCES: A comprehensive search was carried out using PubMed (MEDLINE), Embase, and Web of Science for clinical trials, which studied the usefulness of UT-2 as a diagnostic marker for acute pancreatitis. Sensitivity, specificity and the diagnostic odds ratios (DORs) with 95% confidence interval (CI) were calculated for each study and were compared with serum amylase and lipase. Summary receiver-operating curves were conducted and the area under the curve (AUC) was evaluated. RESULTS: A total of 18 studies were included. The pooled sensitivity and specificity of UT-2 for the diagnosis of acute pancreatitis were 80% and 92%, respectively (AUC=0.96, DOR=65.63, 95% CI: 31.65-139.09). The diagnostic value of UT-2 was comparable to serum amylase but was weaker than serum lipase. The pooled sensitivity and specificity for the diagnosis of post-ERCP pancreatitis were 86% and 94%, respectively (AUC=0.92, DOR=77.68, 95% CI: 24.99-241.48). CONCLUSIONS: UT-2 as a rapid test could be potentially used for the diagnosis of post-ERCP pancreatitis and to an extent, acute pancreatitis. Further studies are warranted to confirm these results.

[Health economic evaluation of clinical pathway of traditional Chinese medicine in the treatment of mild acute pancreatitis].

OBJECTIVE: To evaluate the health economic value clinical pathway (CP) of traditional Chinese medicine in the treatment of mild acute pancreatitis (MAP). METHODS: Ninty one patients with MAP were enrolled prospectively in TCM clinical pathway group from June 2012 to February 2013, while the data of 80 MAP patients who were treated without TCM clinical pathway from June 2011 to May 2012, were analyzed retrospectively as control group. The health economic evaluation data used for the two groups comparison included: average length of stay, hospitalization expenses (total hospitalization expenses, total treatment cost, TCM treatment cost, herbal fees, medicine fees, and nursing care cost), as well as the usage of antibiotics/somatostatin, the release time of abdominal pain, the time of re-feeding, and patient satisfaction. RESULTS: There were no significant statistical differences in demographics, etiology, Ranson and Balthazar CT scores between the two groups (P > 0.05). Compared with non-CP group, the usage of antibiotics and somatostatin, the release time of abdominal pain, the time of re-feeding and patient satisfaction were all improved significantly in CP group (P < 0.05). The average length of stay in CP group was shorter than that of non-CP group (P < 0.05). Total hospitalization expenses [yen (11,089.89 +/- 4,318.29) vs. yen (8,960.34 +/- 4,328.91)], medicine fees [yen (6,563.80 +/- 2,743.87) vs. yen (3,988.28 +/- 2,128.10)] and nursing care cost [yen (110.51 +/- 37.24) vs. yen (93.32 +/- 35.20)] were all reduced in CP group, while TCM treatment cost [yen (609.59 +/- 624.42) vs. (968.29 +/- 769.68)] and herbal fees [yen (162.72 +/- 135.13) vs. yen (303.49 +/- 149.90)] were increased (P < 0.05). There was no significant statistical difference in total treatment cost between the two groups (P > 0.05). CONCLUSION: TCM clinical pathway of MAP can not only ensure the therapeutic effects, but also shorten the average length of stay, reduce medical cost and increase patient satisfaction.

[The value of different scoring systems for computed tomography in predicting pancreatic pseudocyst in severe acute pancreatitis].

OBJECTIVE: To evaluate the value of several Computed Tomograph (CT) scoring systems in predicting the development of acute pancreatic pseudocyst (PPC) in severe acute pancreatitis (SAP) during early One hundred and sixty-two patients with SAP were retrospectively observed and subjected to stage. METHODS clinical, laboratory, and radiology investigation from October 2007 to December 2010. Three different CT scoring systems including CT severity index (CTSI), Modified CT severity index (MCTSI) and Extrapancreatic Inflammation on CT score (EPIC), were used for the determine of PPC, while the predictive values of these three Forty-eight patients CT scoring systems in the presence of PPC were analyzed by the ROC curve. RESULTS: (29.6%) were observed the formation of PPC. The scores of CTSI, MCTSI, EPIC and the occurrence rate of ascites in PPC group were significantly higher than those in non-PPC group with One-way ANOVA analysis. Among the three CT scoring systems, EPIC score showed a larger area under ROC curve (AUC = 0.914) than CTSI (AUC = 0.674) and MCTSI (AUC = 0.72) did. CONCLUSION: EPIC scoring system has better prediction of PPC in SAP patients than CTSI and MCTSI.

Chemical composition and anti-inflammatory activity of flower essential oil from Forsythia koreana Nakai.

Forsythia koreana Nakai is an ornamental plant widely cultivated in East Asia. The essential oil of F. koreana flowers (FEO) was extracted by hydrodistillation process and the volatile components were determined with gas chromatography coupled with mass spectrometry. The anti-inflammatory activity of FEO was investigated by using TPA-induced mouse ear inflammation model. The major components of FEO were identified as n-tetracosane (29.85%), n-heneicosane (17.45%), myristic acid (8.46%) and palmitaldehyde (6.22%). The TPA-induced mouse ear edema, water content, dermis thickness, epidermis thickness and nitric oxide production were decreased by FEO. Our findings suppose that the flower essential oil of F. koreana exerted anti-inflammatory activity, and may be used in the development of anti-inflammatory products in future.

Serum matrix metalloproteinase-9 is an early marker of pancreatic necrosis in patients with severe acute pancreatitis.

BACKGROUND/AIMS: To study the ability of matrix metalloproteinase-9 (MMP-9) to predict pancreatic necrosis (PN) in patients with severe acute pancreatitis (SAP). METHODOLOGY: From July 1, 2010 to December 31, 2010 patients diagnosed with SAP were included (n=35). Serum MMP-9, CRP and IL-6 were analyzed on days 1, 3, 5 and 7 of hospitalization to determine if they could predict the development of pancreatic necrosis. RESULTS: Of the 35 patients included, 12 (34.3%) had evidence of PN. Admission MMP-9 concentrations were significantly higher in patients with PN compared to subjects without PN (13.1±4.0 vs. 7.5±3.8, p<0.05). Receiver operating characteristic curves for PN revealed an area under the curve of 0.832 for admission MMP-9 (95% confidence interval 0.696-0.967, p=0.001). Elevated concentrations of MMP-9 on admission for pancreatic necrosis =9.35mg/L yielded a positive predictive value of 90.9% with a sensitivity of 91.7% and a specificity of 69.6%. Binary logistic regression indicated that MMP-9 was significantly associated with pancreatic necrosis (Odds ratios 25.1, 95% confidence interval 2.7-234.2; p=0.005). CONCLUSIONS: An elevation in serum MMP-9 within the first 24 hours of disease is strongly associated with the development of pancreatic necrosis. This finding may have important clinical implications and requires further investigation.

The study on causes of death in fulminant pancreatitis at early stage and late stage.

OBJECTIVE: To investigate clinical characteristics of fulminant pancreatitis (FP) died at early and late stage, analyze the difference in death causes of FP at these two stage. METHODS: Ninety-two (92) patients with FP were admitted in our center from January 2000 to June 2010, and 55 patients of them died of FP. These dead FP patients were divided into two groups according to the death time: within 7 d (early death group) or after 7 d (late death group). The 24 h Acute Physiology and Chronic Heath Evaluation II (APACHE II) score, the occurrence of complications were compared between these two groups. RESULTS: The mortality of FP was 59.8% (55/92), in which 20.6% (19 cases) died within 3 d and 29.3% (27 cases) died after 14 d. Compared with the late death group, the early death group showed higher 24 h APACHE II score and serum triglyceride level (P < 0.05), and also had higher occurring time of renal failure, shock, hepatic failure, encephalopathy, gastrointestinal hemorrhage and infection (P < 0.05). However, the incidences of encephalopathy, gastrointestinal hemorrhage and pancreatic necrosis infection in the late death group were higher than those in the early group (P < 0.05). In addition, the major pathogenesis of infection was Gram-negative bacterium. CONCLUSION: The most important and common cause of death for the patients with FP is multiple organ dysfunction syndrome, which usually was the consequence of systemic inflammation response syndrome in the early stage, and the severe infection in the later stage, respectively.

[Effect of chai qin cheng qi decoction on serum CCK-8 and calcium overload of pancreatic acinar in mice with acute pancreatitis].

OBJECTIVE: To investigate the effect of Chai Qin Cheng Qi decoction (CQCQD) on serum cholecystokinin-8 (CCK-8) and calcium overload of pancreatic acinar cells in acute pancreatitis (AP) mice. METHODS: Twenty four mice were randomly divided into control group, AP group, CQCQD group and siRNA group, each comprising 6 mice. AP mouse model was induced by intraperitoneal injection of 8% L-arginine in a dose of 4 g/kg. The AP mice in the CQCQD group were fed with 0.4 mL/100 g of Chai Qin Cheng Qi solution once every two hours. The AP mice in the siRNA group were injected intraperitoneally with CCK-siRNA in a dose of 0.88 mg/kg. The changes of serum CCK-8 and calcium concentrations in the pancreatic acinar cells and pancreatic pathology were observed 6 hours after the interventions. RESULTS: The serum CCK-8 [(3764.3 +/- 369.2) ng/mL], calcium fluorescence intensity (34.8 +/- 27.1) of pancreatic acinar cells and pancreas pathology scores (6.2 +/- 1.1) of the AP mice were significantly greater (P < 0.05) than those in the control group [(1253.5 +/- 39.5) ng/mL, 5.2 +/- 2.3, 2.8 +/- 0.4], CQCQD group [(1230.5 +/- 46.1) ng/mL, 9.6 +/- 1.6, 3.8 +/- 0.8, 4.1 +/- 0.5] and siRNA group[(1702.3 +/- 598.3) ng/mL, 7.6 +/- 2.0]. Serum CCK-8 was positively correlated with intracellular calcium concentrations (r = 0.793, P = 0.021) in pancreatic acinar cells and pancreas pathology scores (r = 0.847, P = 0.000). CONCLUSION: Acute pancreatitis in mice induced by L-arginine is associated with calcium overload in pancreatic acinar cells induced by increased serum CCK-8. CQCQD can reduce serum levels of CCK-8, alleviate calcium overload in pancreatic acinar cells, and reduce pancreas pathological changes in AP mice.

Sulfur Regulates the Trade-Off Between Growth and Andrographolide Accumulation via Nitrogen Metabolism in Andrographis paniculata.

Nitrogen (N) and sulfur (S) are essential mineral nutrients for plant growth and metabolism. Here, we investigated their interaction in plant growth and andrographolide accumulation in medicinal plant Andrographis paniculata grown at different N (4 and 8 mmol·L-1) and S concentration levels (0.1 and 2.4 mmol L-1). We found that increasing the S application rate enhanced the accumulation of andrographolide compounds (AGCs) in A. paniculata. Simultaneously, salicylic acid (SA) and gibberellic acid 4 (GA4) concentrations were increased but trehalose/trehalose 6-phosphate (Tre/Tre6P) concentrations were decreased by high S, suggesting that they were involved in the S-mediated accumulation of AGCs. However, S affected plant growth differentially at different N levels. Metabolite analysis revealed that high S induced increases in the tricarboxylic acid (TCA) cycle and photorespiration under low N conditions, which promoted N assimilation and S metabolism, and simultaneously increased carbohydrate consumption and inhibited plant growth. In contrast, high S reduced N and S concentrations in plants and promoted plant growth under high N conditions. Taken together, the results indicated that increasing the S application rate is an effective strategy to improve AGC accumulation in A. paniculata. Nevertheless, the interaction of N and S affected the trade-off between plant growth and AGC accumulation, in which N metabolism plays a key role.

Treatment of functional constipation with the Yun-chang capsule: a double-blind, randomized, placebo-controlled, dose-escalation trial.

BACKGROUND AND AIM: Functional constipation is a common functional bowel disorder for which there is no reliable medical treatment. This study was designed to determine the therapeutic efficacy and safety of the Yun-chang capsule, a Chinese herbal formula, in the treatment of patients with functional constipation. METHODS: In our multi-center, prospective, double-blind, randomized, placebo-controlled, dose-escalation trial, patients with functional constipation received 70 mg of Yun-chang capsule plus 35 mg placebo (group A), 105 mg of Yun-chang capsule (group B), or 105 mg placebo (group C), three times daily for 2 weeks. The primary end-points were the changes in main symptom score and cumulative symptom score 2 weeks after the treatment. The secondary end-points were adverse events. RESULTS: A total of 140 patients were recruited and 132 met the inclusion criteria; 44 patients constituted each of the three treatment groups. Compared with patients in group C, patients in groups A and B had significant improvement in the main symptom score, cumulative symptom score, the change from baseline of the main symptom score, and the change from baseline of the cumulative symptom score at week 1 and week 2. The scores showed slight superiority of group B over group A at week 1 and week 2, although these differences were not statistically significant. There were no differences in adverse events. CONCLUSIONS: The Yun-chang capsule is efficacious and safe for the treatment of patients with functional constipation. Larger and longer-term trials are required to fully assess the benefits and safety of this treatment for functional constipation.

[Establishment of target genomic DNA capturing system for next generation sequencing].

The motivation of this research is to establish a system of target genomic DNA capture and enrichment, which could be used in deep sequencing of target regions with next-generation sequencing. To design the 120 bp capture probes (baits) and prepare the SureSelect reagents, 2,414,977 bp human genomic sequence of 11,824 exons in 1250 genes were submitted to the Agilent eArray platform and manufactured by Agilent. Two human genomic DNA samples were used and conducted the successive experiments for sequencing library construction: shearing fragmentation by sonication, blunt-ending and phosphorylation, adaptor ligation, 150-200 bp fragments size selection, followed by hybridization with the baits, hybrid selection with magnetic beads, and PCR amplification. Prior to SOLiD sequencing reaction, the libraries were amplified with emulsion PCR and enriched with the P2 enrichment beads. The library samples were loaded to sequencing Chip for Work Flow Analysis (WFA) or sequencing running with default parameters. The results displayed that 46 509 baits were designed and synthesized for 11,147 gene regions, and SureSelect capture probe regent was prepared. Real-time PCR showed the target enrichment efficiency up to 2(9) times with the SureSelect system. WFA revealed that the libraries were suitable for SOLiD Sequencing. The sequencing data revealed that 70% of the unique mapped sequence tags matched the target regions, and the average coverage of the target regions were above 200-fold. All these demonstrated the feasibility of the established system of target genome sequence capture for next generation DNA sequencing.

Skeletal muscle-derived cells repair peripheral nerve defects in mice.

Skeletal muscle-derived cells have strong secretory function, while skeletal muscle-derived stem cells, which are included in muscle-derived cells, can differentiate into Schwann cell-like cells and other cell types. However, the effect of muscle-derived cells on peripheral nerve defects has not been reported. In this study, 5-mm-long nerve defects were created in the right sciatic nerves of mice to construct a peripheral nerve defect model. Adult female C57BL/6 mice were randomly divided into four groups. For the muscle-derived cell group, muscle-derived cells were injected into the catheter after the cut nerve ends were bridged with a polyurethane catheter. For external oblique muscle-fabricated nerve conduit and polyurethane groups, an external oblique muscle-fabricated nerve conduit or polyurethane catheter was used to bridge the cut nerve ends, respectively. For the sham group, the sciatic nerves on the right side were separated but not excised. At 8 and 12 weeks post-surgery, distributions of axons and myelin sheaths were observed, and the nerve diameter was calculated using immunofluorescence staining. The number, diameter, and thickness of myelinated nerve fibers were detected by toluidine blue staining and transmission electron microscopy. Muscle fiber area ratios were calculated by Masson's trichrome staining of gastrocnemius muscle sections. Sciatic functional index was recorded using walking footprint analysis at 4, 8, and 12 weeks after operation. The results showed that, at 8 and 12 weeks after surgery, myelin sheaths and axons of regenerating nerves were evenly distributed in the muscle-derived cell group. The number, diameter, and myelin sheath thickness of myelinated nerve fibers, as well as gastrocnemius muscle wet weight and muscle area ratio, were significantly higher in the muscle-derived cell group compared with the polyurethane group. At 4, 8, and 12 weeks post-surgery, sciatic functional index was notably increased in the muscle-derived cell group compared with the polyurethane group. These criteria of the muscle-derived cell group were not significantly different from the external oblique muscle-fabricated nerve conduit group. Collectively, these data suggest that muscle-derived cells effectively accelerated peripheral nerve regeneration. This study was approved by the Animal Ethics Committee of Plastic Surgery Hospital, Chinese Academy of Medical Sciences (approval No. 040) on September 28, 2016.