Differential distribution of characteristic constituents in peel and pulp of Aurantii Fructus Immaturus (Citrus aurantium L.) using MALDI mass spectrometry imaging.

Aurantii Fructus Immaturus (AFI) was structurally divided into two parts named "peel" and "pulp". The exocarp and mesocarp of materials named "peel". The endocarp separated into multiple compartments and the cystic hair attached to it named "pulp". In order to explore the distribution and content of constituents in AFI, an efficient method to explore the distribution of constituents was developed based on matrix assisted laser desorption/ionization fourier transform ion cyclotron resonance mass spectrometry imaging (MALDI-FTICR-MSI). After simple processing, thirty-two constituents with distinct localization in the mass range of 101-1200 Da were identified by MALDI-FTICR-MSI. In addition, the identified four flavnoids (poncirin, sinensetin, 3,5,6,7,8,3',4'-heptemthoxyflavone, and tangeritin) were analyzed for differences between using LC-MS. Quantitative analysis results supported the quantitative results from MALDI-FT-ICR-MSI. The results implied that different parts had different constituents in AFI, and demonstrated MALDI-MSI have high potential in the direct analysis of constituents.

CYP72D19 from Tripterygium wilfordii catalyzes C-2 hydroxylation of abietane-type diterpenoids.

KEY MESSAGE: CYP72D19, the first functional gene of the CYP72D subfamily, catalyzes the C-2 hydroxylation of abietane-type diterpenoids. The abietane-type diterpenoids, e.g., triptolide, tripdiolide, and 2-epitripdiolide, are the main natural products for the anti-tumor, anti-inflammatory, and immunosuppressive activities of Tripterygium wilfordii, while their biosynthetic pathways are not resolved. Here, we cloned and characterized the CYP72D19-catalyzed C-2 hydroxylation of dehydroabietic acid, a compound that has been proven to be a biosynthetic intermediate in triptolide biosynthesis. Through molecular docking and site-directed mutagenesis, L386, L387, and I493 near the active pocket were found to have an important effect on the enzyme activity, which also indicates that steric hindrance of residues plays an important role in function. In addition, CYP72D19 also catalyzed a variety of abietane-type diterpenoids with benzene ring, presumably because the benzene ring of the substrate molecule stabilized the C-ring, allowing the protein and the substrate to form a relatively stable spatial structure. This is the first demonstration of CYP72D subfamily gene function. Our research provides important genetic elements for the structural modification of active ingredients and the heterologous production of other 2-hydroxyl abietane-type natural products.

Patellar instability-induced bone loss in the femoral trochlea is associated with the activation of the JAK1/STAT3 signaling pathway in growing mice.

INTRODUCTION: Patellar instability (PI) at an early age is believed closely correlated with bone loss in the development of the femoral trochlea and can cause trochlear dysplasia. However, the molecular mechanism of PI-induced bone loss has not been established. The Janus kinase (JAK)/signal transducers and activators of transcription (STAT) signaling pathway plays an important role in bone development by regulating the expression of osteoprotegerin (OPG) and receptor activator of nuclear factor kappa B ligand (RANKL). The aim of this study was to explore the association of JAK1/STAT3 signaling to PI-induced subchondral bone loss in the femoral trochlea. METHODS: Four-week-old male C57BL/6 mice were randomly divided into two groups (n = 50/group). Mice in the experimental group underwent surgery to induce PI. Distal femurs were collected 2 and 4 weeks after surgery (n = 25 knees/each time point, each group). Microcomputed tomography and histological observations were performed to investigate the morphology of the femoral trochlea and changes in bone mass. qPCR, western blot, and immunohistochemistry analyses were performed to evaluate the expression of JAK1, STAT3, RANKL, and OPG in subchondral bone. A t test was performed for the statistical analysis; a P value < 0.05 was considered to be statistically significant. RESULTS: In the experimental group, subchondral bone loss in the femoral trochlea was observed two and four weeks after PI; morphological changes, such as a flatter trochlear groove and an increased sulcus angle, were observed in the femoral trochlea; qPCR, western blot, and immunohistochemistry analyses showed higher expression of JAK1, STAT3, and RANKL and lower expression of OPG (P < 0.05). CONCLUSION: PI-induced subchondral bone loss in the femoral trochlea and resulted in trochlear dysplasia in growing mice. This bone loss is associated with activation of the JAK1/STAT3 signaling pathway, which weakens the function of osteoblasts and stimulates both formation and function of osteoclasts.

Electrophysiological and pathological changes in the vastus medialis and vastus lateralis muscles after early patellar reduction and nerve growth factor injection in rabbits with patellar dislocation.

BACKGROUND: Patellar dislocation can cause a series of changes in the trochlear groove and patella. However, the influence of patellar dislocation on the medialis (VM) and vastus lateralis (VL) muscles and whether nerve growth factor (NGF) is beneficial to proprioceptive rehabilitation for patellar dislocation are unknown. The purpose of this study was to investigate the effects on VM and VL after the injection of NGF and early reduction in rabbits for patellar dislocation with electrophysiological and pathological analysis. METHODS: Sixty 2-month-old rabbits were randomly divided into four groups (15 rabbits in each group). Rabbits in Group 1, Group 2, and Group 3 underwent patellar dislocation surgery, and rabbits in Group 4 underwent sham surgery. One month later, patellar reduction was performed in Groups 1 and 2. NGF was injected into the rabbits of Group 1. The electrophysiological and pathological changes in VM and VL were analyzed at 1 month and 3 months after patellar reduction. RESULTS: The electrophysiological and pathological indices in Groups 1 and 2 were significantly different from those in Group 3 at 1 and 3 months after patellar reduction. There were significant differences between NGF injection Group 1 and Group 2 without NGF injection. There was no significant difference between Group 1 and Group 4 at 3 months after patellar reduction. CONCLUSIONS: Patellar dislocation can cause abnormal electrophysiological and pathological effects on VM and VL. Patellar reduction should be performed as early as possible, and NGF injection may be beneficial to the rehabilitation of proprioception.

Suppressive effects of RASAL2 on renal cell carcinoma via SOX2/ERK/p38 MAPK pathway.

Metastatic renal cell carcinoma (RCC) is associated with poor prognosis. Ras protein activator like 2 (RASAL2) protein has been previously demonstrated to serves as a tumor suppressor in a variety of malignancies. Therefore, the aim of the present study was to investigate the role of RASAL2 in RCC. Reverse transcription-quantitative PCR, western blot analysis and immunohistochemistry were performed to measure mRNA and protein expression in RCC tissues, whilst immunofluorescence and western blotting were performed to evaluate protein expression in RCC cells. A Cell Counting Kit-8 and 5-bromo-2'-deoxyuridine staining were applied to determine cell viability, and Transwell assays were conducted to measure RCC cell invasion and migration. RASAL2 expression was identified to be downregulated in RCC tissues, which associate negatively with RCC pathological grade. Sox2 expression, in addition to ERK1/2 and p38 MAPK phosphorylation, were demonstrated to be increased in RCC tissues. In RCC cells, RASAL2 overexpression decreased the expression of Sox2 and the activation of ERK1/2 and p38 MAPK. Physiologically, RASAL2 overexpression decreased RCC cell viability, invasion and migration. The expression of metalloproteinase-2/9 and tissue inhibitor of metalloproteinase 1 were also identified to be decreased and increased by RASAL2 overexpression, respectively. By contrast, RASAL2 knockdown exerted opposite effects on RCC cells compared with those observed following RASAL2 overexpression. RASAL2 expression decreased RCC cell viability, migration and invasion, which was demonstrated to be associated with the inactivation of SOX2/ERK1/2/p38 MAPK signaling. These results suggest that RASAL2 may potentially serve as a potential target for the development of novel therapeutic intervention strategies against RCC.

The safety and efficiency of retroperitoneal laparoscopic adrenalectomy via extra and intra perinephric fat approaches: a retrospective clinical study.

BACKGROUND: This retrospective clinical study is to evaluate the safety and efficiency of two different approaches in retroperitoneal laparoscopic adrenalectomy and provide experience and basis for the treatment of adrenal tumors through retroperitoneal approach. METHODS: From July 2015 to February 2018, 112 patients with adrenal lesions underwent retroperitoneal laparoscopic adrenalectomy (RLA) using a 3-port method. Among them, 56 patients underwent RLA via the extra perinephric fat approach (EPFA), 56 patients underwent RLA via the intra perinephric fat approach (IPFA). Clinical data, including preoperative, operative and postoperative management were recorded. RESULTS: All surgeries were successfully completed, and there was no single patient who died during these surgeries. There was no statistically significant difference between the two groups in blood loss, postoperative complications, vena cava injury, renal cortex injury, peripheral organ injury, and post operation hospital stay. Peritoneum injury occurred more frequently in the EPFA group when compared with the IPEA group (p = 0.042). The average surgery time of the IPEA group is significantly shorter when compared with that of the EPEA group (p < 0.001). Due to serious saponification of the perinephric fat and heavy adhesion to renal fascia, three cases in IPFA group were converted to the EPFA surgery. CONCLUSION: RLA is a safe and effective procedure both via extra perinephric fat and intra perinephric fat approaches. IPEA is superior to EPEA in terms of peritoneal injury and duration. The choice may mainly depend on the experience of the surgeon, the characteristics of the adrenal tumor and the nature of the perinephric fat.

FGF5 promotes osteosarcoma cells proliferation via activating MAPK signaling pathway.

Objective: This study aimed to investigate the role of fibroblast growth factor-5 (FGF5) in osteosarcoma (OS) and explore the potential mechanisms. Methods: OS gene expression data was downloaded from the Gene Expression Omnibus (GEO; GSE12865) and analyzed by R software. OS tissues and cell lines were collected. The expression level of FGF5 in tumor tissues and cell lines was detected using qRT-PCR. Knockout of FGF5 was performed using CRISPR/Cas9 system. The effects of FGF5 knockout on OS cell proliferation and tumor growth were determined through cell counting kit-8 assay and xenograft nude mice, respectively. Additionally, recombinant FGF5 (rFGF5) was added into OS cell and the effects of rFGF5 on the proliferation and apoptosis of OS cell lines were assayed. Furthermore, the protein expression levels of mitogen-activated protein kinase (MAPK) signaling pathway were detected through Western blot. Results: FGF5 was significantly upregulated in OS tissues and cells, and closely associated with poor differentiation, larger tumor size, lymph node metastasis, and advanced TNM stage. FGF5 knockout could inhibit proliferation of OS cells and tumor growth in nude mouse model. Addition of exogenous rFGF5 promoted OS cell proliferation while inhibited OS cell apoptosis. The expression levels of MAPK signaling pathway proteins in FGF5 knockout group were significantly lower than that in control when there was no rFGF5. Additionally, their expression level in rFGF5 addition group was higher than that in without rFGF5 group. Conclusion: We demonstrated for the first time that FGF5 was overexpressed in OS cell lines and clinical tissue samples and promotes OS cell proliferation by activating MAPK signaling pathway, which indicated that FGF5 was a potential therapeutic target for OS.

Non-needle acupoint stimulation for prevention of nausea and vomiting after breast surgery: A meta-analysis.

BACKGROUND: Breast disease has been a global serious health problem, among women. Surgery is the main treatment for the patients suffering from breast disease. Postoperative nausea and vomiting are still disturbing. Acupoint stimulation, an effective treatment of traditional Chinese medicine, has been used to reduce postoperative nausea and vomiting. Recently, non-needle acupoint stimulation becomes a new intervention. Though several clinical trials have been done, there is still no final conclusion on the efficacy. This Meta-Analysis aims at evaluating the efficacy of non-needle acupoint stimulation for prevention of nausea and vomiting after breast surgery. METHODS: Systematic searches were conducted in PubMed, Embase, Cochrane, and Wanfang Med Online databases for studies. The review period covered from the inception of databases to December 31, 2017. The outcome measures of interest were frequency of nausea, frequency of vomiting, frequency of PONV, verbal rating scale of nausea, and use of rescue antiemetic. Data extraction and risks of bias evaluation were accomplished by 2 independent reviewers using the Cochrane Collaboration Review Manager software (RevMan 5.3.5). RESULTS: Fourteen randomized controlled trials with a total of 1009 female participants in the non-needle acupoint stimulation group and control group met the inclusion criteria. Although the therapeutically effect on vomiting within postoperative 2 hours was not obvious, non-needle acupoint stimulation still had an important role in reducing nausea and vomiting within postoperative 48 hours. According to Jadad scale, there was moderate quality evidence for the pooled analysis results in this study. In addition, stimulating acupoint by wristband acupressure was more likely to cause adverse reactions. CONCLUSION: Non-needle acupoint stimulation can be used for female patients undergoing breast surgery to reduce postoperative nausea and vomiting. Into consideration, we recommend transcutaneous acupoint electrical stimulation on PC6 from 30 minutes before induction of anesthesia to the end of surgery for application. This non-pharmaceutical approach may be promising to promote the recovery of patients after breast surgery.

A review of radiofrequency ablation: Large target tissue necrosis and mathematical modelling.

Radiofrequency ablation (RFA) is an effective clinical method for tumour ablation with minimum intrusiveness. However, the use of RFA is mostly restricted to small tumours, especially those <3cm in diameter. This paper discusses the state-of-the-art of RFA, drawn from experimental and clinical results, for large tumours (i.e. ⩾3cm in diameter). In particular, the paper analyses clinical results related to target tissue necrosis (TTN) and mathematical modelling of the RFA procedure to understand the mechanism whereby the TTN is limited to under 3cm with RFA. This paper also discusses a strategy of controlling of the temperature of target tissue in the RFA procedure with the state-of-art device, which has the potential to increase the size of TTN. This paper ends with a discussion of some future ideas to solve the so-called 3-cm problem with RFA.

Comparing the Diagnostic Potential of 68Ga-Alfatide II and 18F-FDG in Differentiating Between Non-Small Cell Lung Cancer and Tuberculosis.

UNLABELLED: The objectives of this study were to compare the diagnostic potential of (68)Ga-Alfatide II with(18)F-FDG in differentiating between non-small cell lung cancer patients (NSCLC) and lung tuberculosis (TB) patients. METHODS: Twenty-one NSCLC patients and 13 TB patients were recruited. PET/CT images using either (68)Ga-Alfatide II or (18)F-FDG were acquired in 2 consecutive days. SUV quantitative comparison, receiver-operating curve analysis, and comprehensive visual analysis were performed. The expression of the angiogenesis marker αvß3 in NSCLC and TB primary lesions was analyzed by immunohistochemistry. RESULTS: The (68)Ga-Alfatide II SUVmax and SUVmean were significantly different in NSCLC and TB (P = 0.0001 and 0.0007, respectively). The area under the receiver-operating curve value of (68)Ga-Alfatide II SUVmax was significantly higher than that of (18)F-FDG (P = 0.038). The visual differentiation diagnostic specificity of (68)Ga-Alfatide II was 1.57-fold (84.62% vs. 53.85%) higher than that of (18)F-FDG. In the detection of NSCLC lymph nodes, (68)Ga-Alfatide II was superior in specificity (100% vs. 66.7%), whereas the sensitivity was greater with (18)F-FDG (87.5% vs. 75%). In TB lymph node detection, the false-positive rate of (68)Ga-Alfatide II was one-third (15.4%/46.2%) the value of (18)F-FDG. Additionally, (68)Ga-Alfatide II detected more metastases in the brain but less in the liver and the bone. The αvß3 biomarker was specifically expressed in the cells and the neovasculature of NSCLC lesions. CONCLUSION: (68)Ga-Alfatide II is qualified for detecting NSCLC primary lesions and is superior to (18)F-FDG in distinguishing NSCLC from TB in primary lesions and suggestive lymph nodes. (68)Ga-Alfatide II is more likely to be capable of detecting brain metastasis, and (18)F-FDG is more likely to be capable of detecting liver and early-stage bone metastases.

Enteral nutrition is superior to total parenteral nutrition for pancreatic cancer patients who underwent pancreaticoduodenectomy.

OBJECTIVE: To determine the effects of total parenteral nutrition (TPN) and enteral nutrition (EN) on biochemical and clinical outcomes in pancreatic cancer patients who underwent pancreaticoduodenectomy. METHODS: From the year 2006 to 2008, 60 patients who underwent pancreaticoduodenectomy in Tianjin Third Central Hospital were enrolled in this study. They were randomly divided into the EN group and the TPN group. The biochemical and clinical parameters were recorded and analyzed between the two groups. RESULTS: There was no significant difference in the nutritional status, liver and kidney function, and blood glucose levels between the TPN and EN groups on the preoperative day, the 1st and 3 rd postoperative days. However, on the 7th postoperative day, there was significant difference between the two groups in 24 h urinary nitrogen, serum levels of, total protein (TP), transferrin (TF), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and γ-glutamyl transpeptadase (GGT), blood urea nitrogen (BUN) and creatinine (Cr). On the 14th postoperative day, there was a significant difference between the two groups in terms of urinary levels of 24 h nitrogen, TP, TF, retinol binding protein, ALT, AST, ALP, GGT, total bilirubin, direct bilirubin, BUN, Cr, and glucose. The incidence of delayed gastric emptying in the EN and TPN groups was 0% and 20%, respectively. Moreover, the incidence of pancreatic fistulas and hemorrhages in the EN group were 3.6% and 3.6%, versus 26.7% and 30% in the TPN group, respectively. CONCLUSIONS: EN is better than TPN for pancreatic cancer patients who received pancreaticoduodenectomy.

Evaluation of the performance of the EIAgen HCV test for detection of hepatitis C virus infection.

The EIAgen HCV test (Adaltis Inc., Montreal, Canada) is an enzyme immunoassay (EIA) for the detection of anti-hepatitis C virus (HCV) antibodies. This study compared the performance of this test side-by-side with the current Ortho HCV 3.0 Anti-HCV assay (Ortho-Clinical Diagnostics Inc., Johnson & Johnson Company, Raritan, NY, USA). Among 2559 specimens examined, 178 were true positives, 2376 were true negatives and 5 were indeterminate. The sensitivity of the EIAgen HCV test was 100%, versus 98.3% for the Ortho HCV test, while their respective specificities were 98.1% and 98.2%. The EIAgen HCV test gave a positive predictive value of 79.8% and a negative predictive value of 100%. Overall, the concordance of this test with the Ortho HCV test was 98.2%. Specimens from potentially interfering substances, such as sera from pregnant women, sera from patients with acute non-C hepatitis, autoimmune diseases, lipidemia, or from patients undergoing hemolysis, showed no interference with either EIA. An EIAgen HCV test signal-to-cut-off ratio of >5.9 would be highly predictive of a true-positive finding in these specimens. The EIAgen HCV test is well suited for screening blood and blood products in antibodies to HCV.

Bile tract adenomyoma: a case report.

This paper described a rare case of adenomyoma of common bile duct. The case is a 51-year-old man who was hospitalized for yellow color skin and sclera and itching for 2 mo without abdominal pain. Nothing special was found in physical examination except yellowish skin and sclera. The clinical presentation and Computerized Tomography (CT), Magnetic resonance cholangiopancreatography (MRCP), and ultrasonography suspected a tumor of the distal bile duct. The patient was treated successfully by pancreaticoduodenectomy. Histologically, the lesion consisted of adenoid and myofibrous tissue and moderate atypia. The immunophenotype of the epithelial component was cytokeratin 7+/cytokeratin 20-. The patient has been well without any evidence of recurrence for 12 mo since his operation.

A multi-Chinese blood center study testing serologic-negative donor samples for hepatitis C virus and human immunodeficiency virus with nucleic acid testing.

BACKGROUND: A multi-blood center study was conducted to evaluate a human immunodeficiency virus type 1 (HIV-1) and hepatitis C virus (HCV) multiplex nucleic acid testing (NAT) donor screening test and to determine the residual risk for HIV-1 and HCV infection. STUDY DESIGN AND METHODS: A commercially available HIV-1 and HCV assay (Procleix, Chiron Corp.) was used for simultaneous detection of HIV-1 RNA and HCV RNA on 89,647 unlinked donor samples. NAT was performed with pools of 16 samples that had passed all routine screening tests. Single-donor NAT was performed for samples that had been disqualified by any reactive screening test result(s). Anti-HCV (Ortho third-generation HCV enzyme immunoassay [EIA]), alanine aminotransferase, and HCV NAT (Roche COBAS Amplicor HCV test) confirmatory tests were used for HCV EIA-nonreactive, HCV NAT-reactive samples. RESULTS: Three HCV NAT yield cases and no HIV-1 yield cases were detected. The yield rate for HCV NAT was 3.4 per 10(5) (95 percent confidence interval [CI], 0.7-9.8). The estimated incidence rate for HCV is 24.2 per 100,000 person-years (95% CI, 3.4-88.0). If minipool NAT is added to routine donor screening, the residual risk for HCV is estimated to be reduced to 1 in 20.4x10(4) (95% CI, 1 in 5.2x10(4)-1 in 165.5x10(4)). CONCLUSION: The residual risk for transfusion-transmitted HCV infection is still relatively high in China. Incorporating NAT technology into blood donor screening would be estimated to reduce the residual risk of HCV infections eightfold over current EIA screening.

Significance of the signal-to-cutoff ratios of anti-hepatitis C virus enzyme immunoassays in screening of Chinese blood donors.

BACKGROUND: The correlation between signal-to-cutoff (S/CO) ratios of a second-generation hepatitis C virus (HCV) enzyme immunoassay (EIA; Abbott) and a third-generation HCV enzyme-linked immunosorbent assay (ELISA; Ortho) and confirmed HCV infection has been reported. The utility of the values for the Chinese anti-HCV EIA kits, however, has not been studied in evaluating test results in Chinese blood donors. STUDY DESIGN AND METHODS: A total of 156 donor samples repeat reactive for anti-HCV at routine screening from five representative regions of China were retested for anti-HCV by the Ortho third-generation HCV ELISA and six Chinese EIA kits and for HCV RNA by a human immunodeficiency virus-1 and HCV assay (Procleix, Chiron Corp.). The HCV RNA-nonreactive samples were further tested for anti-HCV by a third-generation recombinant immunoblot assay RIBA (Chiron Corp.). The positive result by either nucleic acid amplification test or RIBA was interpreted as confirmed HCV infection. RESULTS: The confirmed HCV prevalence rate in donors in five representative regions obtained in this study was 0.20 percent (77/37,900) in 2004. All seven anti-HCV EIA kits had a significant correlation between S/CO ratios and confirmed HCV infection. The threshold S/CO ratios, which predicted more than 95 percent of confirmed HCV infections for the Ortho, SABC, BGI-GBI, InTec, GWK, KHB, and WANTAI kits, were 3.8, 6.0, 7.0, 8.6, 10.0, 10.0, and 14.0, respectively. CONCLUSIONS: Anti-HCV EIA kits commonly used in Chinese donors screening demonstrate good correlation between S/CO ratios and the confirmed infection. For the Ortho third-generation HCV ELISA, the S/CO ratio of 3.8 determined by the US Centers for Disease Control and Prevention is applicable to Chinese blood donors. The Chinese domestic EIA kits evaluated show a diverse range of threshold S/CO ratios.

[Correlation between signal/cutoff ratios of anti-HCV enzyme immunoassay (EIA) and their true positivity in blood donors].

OBJECTIVES: To evaluate the correlation between signal/cutoff (S/CO) ratios of anti-HCV EIA and their true positivity for determining the predictive value of S/CO ratios. METHODS: One hundred and fifty-nine samples of blood from donors positive for anti-HCV at the initial screening were collected from Beijing, Guangzhou, Hangzhou, Kunming and Urumchi. All the samples were retested by Ortho and 6 Chinese domestic anti-HCV EIA kits in duplicate, and detected for HCV RNA (NAT) using Chiron Procleix HIV/HCV system (transcription mediated amplification, TMA). The HCV RNA negative samples were further tested for anti-HCV by Chiron RIBA 3.0. Either NAT or RIBA positive samples were interpreted as the true positive. RESULTS: All 7 anti-HCV EIA kits had a significant correlation between S/CO ratios and true positivity. The S/CO ratio of Ortho > or = 3.8 predicted the true positivity in 96.1% of the samples tested. The S/CO ratios of BGI-GBI, GWK, SABC, KHB, InTec, and Wantai were > or = 7.0, > or = 10.0, > or = 6.0, > or = 10.0, > or = 8.6, > or = 14.0 and predicted 96.1%, 96.1%, 97.3%, 96.0%, 96.1%, 96.0% of the true positivity, respectively. CONCLUSIONS: The S/CO ratios of anti-HCV EIA kits are associated with the true positivity. S/CO ratios of Ortho, BGI-GBI, GWK, SABC, KHB, InTec and Wantai predicting > or = 95% true positivity are > or = 3.8, > or = 7.0, > or = 10.0, > or = 6.0, > or = 1 0.0, > or = 8.6 and > or = 14.0, respectively.