Objective To investigate the metabolism characteristics and the potetial biomarker candidates of osteonecro?sis of the femoral head (ONFH) using metabolomic technology. Methods The femoral head specimens from 23 ONFH patients (25 necrotic femoral heads) and 18 normal femoral heads from femoral neck fracture patients were collected for histopathological examination to confirm the diagnosis of all samples. All the metabolites of bone trabecula were extracted for ultra-high perfor?mance liquid chromatography-MS/MS analyzed. The measured variables was pretreat, and PCA (principal component analysis), PLS?DA (partial least squares?discriminant analysis) and OPLS?DA (orthogonal?partial least squares?discriminant analysis) models were employed to confirm the difference between these two groups after UPLC?MS/MS (ultra?high performance liquid chromatogra?phy?mass spectrometry/mass spectrometry) analysis. At last, the differential variables were screened out by PLS?DA and variate analysis (Kruskal?Wallis H test). The changed metabolites were confirmed by MS and MS/MS aligned in HMDB (human metabolo?mic database) and Massbank. The changed metabolites with the most obviously changed peak abundance, D?arginine, L?proline and L?glutamine, were picked out as the potential diagnostic biomarkers. After binary logistic regression analysis, the combined biomarkers candidates were further analyzed by receiver operating characteristic (ROC) curve to evaluate the significance of the combined biomarkers. Results Significant distinction of metabolites expression mode can be seen in PCA, PLS?DA and OPLS?DA models scoring plots between ONFH and control groups. Twelve changed metabolites in ONFH bone trabeculas were con?firmed by multi?variate statistical analysis and variate statistical analysis. Compared with the femoral neck fracture patients, the in?creased metabolites included D?arginine, L?proline, L?glutamine, creatine, uracil, uridine, LysoPC(20∶4(5Z, 8Z, 11Z, 14Z)), Ly?soPC(16∶0), PC(20∶1(11Z)/18∶3(6Z, 9Z, 12Z)) and PE(P?16∶0e/0∶0). The decreased metabolites were reticulataxanthin and β?cryptoxanthin. According to the change fold of peak abundance and variable weight projection in PLS?DA, the most obviously dif? ferential metabolites were picked out as the biomarker candidates of ONFH. The potential biomarkers candidates were identified as D?arginine, L?proline and L?glutamine. The area under the curve of D?arginine, L?proline and L?glutamine ROC were 0.873, 0.712 and 0.862. The area under the curve of ROC was 0.946 after combining D?arginine, L?proline, L?glutamine using binary lo?gistic regression analysis. Conclusion PCA, PLS?DA and OPLS?DA models were used to find out the differential variables in the metabolites of bone trabeculas in ONFH and femoral neck fracture patients. Twelve metabolites were identified by MS/MS, and 3 obviously changed metabolites, D?arginine, L?proline, L?glutamine, were indicated as biomarker candidates. These 3 obviously changed metabolites showed a good diagnostic significance.

Objective To study the distribution rule of sense and motor fibers in the normal trigeminal nerve roots by electrophysiologic monitoring.Methods Twenty patients accepted microvascular decompression because of primary mimetic convulsion, collected in our hospital from January 2011 and December 2013, was studied their normal trigeminal nerve roots through electroneurophysiology;during the operation, the trigeminal nerve roots were divided into 8 areas: Ⅰ area is the external lateral area, Ⅱ area is the external upper area, Ⅲ area is the middle upper area, Ⅳ area is the internal upper area, Ⅴ area is the internal lateral area, Ⅵ area is the internal bottom area, Ⅶ area is the middle bottom area, and Ⅷ area is the external bottom area;and then, one mm diameter bipolar stimulator was adopted to give 0.2 mA electrical stimulation on these areas, respectively;and the compound nerve action potential (CNAP) at the foramina supraorbitale (V1), foramina infraorbitale (V2) and foramina mentale (V3) was recorded as the CNAPV1, CNAPV2 and CNAPV3 respectively;the compound muscle action potential (CMAP) at the masseter muscle (MM) and temporal muscle (TM) was recorded as CMAPMM and CMAPTM respectively;and finally, the distribution rules of the sense and motor fibers inside the trigeminal nerve roots were judged according to the laws of CNAP and CMAP emergence.Results During the operation, all patients could get stable CNAP and CMAP wave patterns at periphery according to the electrical stimulation on trigeminal nerve roots;the CNAPV1, CNAPv2 and CNAPV3, CMAPMM and CMAPTM seldom appeared in the Ⅰ, Ⅱ and Ⅷ areas of trigeminal nerve roots, but more often appeared in the Ⅳ, Ⅴ and Ⅵ areas, enjoying significant differences (P＜0.05);motor fiber located in the internal lateral area of the nerve, and the sense fiber located in the middle and external lateral area.By stimulating the sense taproot fiber, CNAP could be recorded, and by stimulation the motor taproot fiber, CMAP and several CNAP from V1, V2 and V3 could be recorded.Conclusion Electrical stimulation on motor taproot implies that there are anastomosing fibers existting in both motor and sense fibers;according the 8 areas being divided, motor fiber locating in the internal lateral area of the nerve, and sense fibers locating in the middle and external lateral areas can be concluded.

OBJECTIVETo develop a new method to rapidly determine and identify Guizhi Fuling capsule by portable acousto-optic tunable filter-near infrared spectroscopy.

METHODThe qualitative model was set up using principal component analysis. The correlation models between the NIR spectra and the reference values of five major constituents were obtained with partial least squares method.

RESULTThe identifying model accurately identified Guizhi Fuling capsule, and quantitative analytical models could precisely predicted the content of ellagic acid, baicalin, benzoylpaenoniflorin, cinnamaldehyde, and paeonol. The correlation coefficients of the calibration models were 0.924 2, 0.938 4, 0.924 2, 0.933 6, 0.934 7, the validation set coefficients of the calibration were 0.924 2, 0.938 4, 0.924 2, 0.933 6, 0.934 7, and the RMSEP were 1.138%, 3.014%, 0.751%, 0.625%, 3.455%, 1.363%, respectively. The results of external validation showed no significant difference between the predictive and the determining values by t-test.

CONCLUSIONThe method is accurate, rapid and non-destructive, and can be used for determining and identifying Guizhi Fuling capsule.

Acetophenones ; analysis ; Acrolein ; analogs & derivatives ; analysis ; Calibration ; Capsules ; chemistry ; Drug Evaluation ; methods ; Drugs, Chinese Herbal ; analysis ; Ellagic Acid ; analysis ; Flavonoids ; analysis ; Least-Squares Analysis ; Models, Chemical ; Principal Component Analysis ; methods ; Spectroscopy, Near-Infrared ; methods

OBJECTIVETo analyze the relationship between hemomyelogram and sererity of acute appendicitis and identify the best routine blood test feature for perforation.

METHODS721 patients were included in this study, all of whom underwent appendectomy for the clinical diagnosis of appendicitis during the years of 2010-2013. The initial preoperative hemomyelogram was evaluated at different stages of appendicitis. The area under the ROC curve was used to assess the clinical feature with greater diagnostic accuracy of perforation. Total lymphocyte counts of 1.83 was used in the prediction of perforative appendicitis. A group of 467 patients was used for validation to confirm the diagnostic value of the cut-off value.

RESULTSThe percentage of lymphocytes had the closest association with the evolutionary phase of acute appendicitis. Total lymphocyte counts < or=1.83 indicated perforation, with high sensitivity and low specificity.

CONCLUSIONThe percentage of lymphocytes and total lymphocyte counts are helpful as a diagnostic paramete for different stages of acute appendicitis.

Acute Disease ; Appendectomy ; Appendicitis ; diagnosis ; Humans ; Lymphocyte Count ; ROC Curve ; Sensitivity and Specificity

Objective To analyze the relationship between hemomyelogram and sererity of acute appendicitis and identify the best routine blood test feature for perforation. Methods 721 patients were included in this study,all of whom underwent appendectomy for the clinical diagnosis of appendicitis during the years of 2010- 2013. The initial preoperative hemomyelogram was evaluated at different stages of appendicitis. The area under the ROC curve was used to assess the clinical feature with greater diagnostic accuracy of perforation. Total lymphocyte counts of 1.83 was used in the prediction of perforative appendicitis. A group of 467 patients was used for validation to confirm the diagnostic value of the cut-off value. Results The percentage of lymphocytes had the closest association with the evolutionary phase of acute appendicitis. Total lymphocyte counts < or=1.83 indicated perforation, with high sensitivity and low specificity. Conclusion The percentage of lymphocytes and total lymphocyte counts are helpful as a diagnostic paramete for different stages of acute appendicitis.

Objective The aim of this study was to investigate the effects of Rad9 mutants with impaired DNA mismatch repair ( MMR) function on the tumorigenesis of colorectal cancer. Methods The colorectal cancer tumor samples were collected from 100 patients. The mutation profiles of human Rad9 ( hRad9) gene in these samples were detected by reverse transcriptase?polymerase chain reaction ( RT?PCR) and sequencing. The plasmid of pFLAG?hRad9 ( L101M ) was constructed following the QuickChange mutagenesis procedure and transfected into mRad9?deleted mouse cells ( mRad9-/- cells) . The expression of hRad9 protein was measured by western blot analysis. The MMR activity in live cells was detected by flow cytometry using the reporter plasmid for MMR function. Results Mutation from Leu to Met at the residue 101 ( L101M) of hRad9 gene was detected in 7 of the 100 samples. The mismatch repair efficiency of mRad9-/-+L101M cells ( mRad9?deleted mouse cells with ectopic expression of L101M hRad9 gene) was (34.0± 5. 6)%, which was significantly lower than that in the mRad9-/-+ hRad9 cells [ mRad9?deleted mouse cells with ectopic expression of hRad9 gene, (48.0±7.5)%, P<0.05]. After N?nitroso?N?methylurea (MNU) treatment, the survival rate of mRad9-/-+L101M cells was (33.7±5.9)%, which was significantly higher than that in the mRad9-/-+ hRad9 cells [(21.3±4.7)%, P<0.05]. Thus, ectopic expression of L101M hRad9 gene resulted in significantly reduced MMR activity and increased resistance to MNU. Furthermore, ectopic expression of hRad9 gene with mutation at the target residues of post?translational modification in mRad9-/- cells also led to a reduced MMR activity. Conclusion Rad9 mutants with impaired DNA mismatch repair function may promote tumorigenesis of colorectal cancer.

Objective To analyze the relationship between hemomyelogram and sererity of acute appendicitis and identify the best routine blood test feature for perforation. Methods 721 patients were included in this study,all of whom underwent appendectomy for the clinical diagnosis of appendicitis during the years of 2010- 2013. The initial preoperative hemomyelogram was evaluated at different stages of appendicitis. The area under the ROC curve was used to assess the clinical feature with greater diagnostic accuracy of perforation. Total lymphocyte counts of 1.83 was used in the prediction of perforative appendicitis. A group of 467 patients was used for validation to confirm the diagnostic value of the cut-off value. Results The percentage of lymphocytes had the closest association with the evolutionary phase of acute appendicitis. Total lymphocyte counts < or=1.83 indicated perforation, with high sensitivity and low specificity. Conclusion The percentage of lymphocytes and total lymphocyte counts are helpful as a diagnostic paramete for different stages of acute appendicitis.

Objective The aim of this study was to investigate the effects of Rad9 mutants with impaired DNA mismatch repair ( MMR) function on the tumorigenesis of colorectal cancer. Methods The colorectal cancer tumor samples were collected from 100 patients. The mutation profiles of human Rad9 ( hRad9) gene in these samples were detected by reverse transcriptase?polymerase chain reaction ( RT?PCR) and sequencing. The plasmid of pFLAG?hRad9 ( L101M ) was constructed following the QuickChange mutagenesis procedure and transfected into mRad9?deleted mouse cells ( mRad9-/- cells) . The expression of hRad9 protein was measured by western blot analysis. The MMR activity in live cells was detected by flow cytometry using the reporter plasmid for MMR function. Results Mutation from Leu to Met at the residue 101 ( L101M) of hRad9 gene was detected in 7 of the 100 samples. The mismatch repair efficiency of mRad9-/-+L101M cells ( mRad9?deleted mouse cells with ectopic expression of L101M hRad9 gene) was (34.0± 5. 6)%, which was significantly lower than that in the mRad9-/-+ hRad9 cells [ mRad9?deleted mouse cells with ectopic expression of hRad9 gene, (48.0±7.5)%, P<0.05]. After N?nitroso?N?methylurea (MNU) treatment, the survival rate of mRad9-/-+L101M cells was (33.7±5.9)%, which was significantly higher than that in the mRad9-/-+ hRad9 cells [(21.3±4.7)%, P<0.05]. Thus, ectopic expression of L101M hRad9 gene resulted in significantly reduced MMR activity and increased resistance to MNU. Furthermore, ectopic expression of hRad9 gene with mutation at the target residues of post?translational modification in mRad9-/- cells also led to a reduced MMR activity. Conclusion Rad9 mutants with impaired DNA mismatch repair function may promote tumorigenesis of colorectal cancer.

OBJECTIVE： To screen active fractions of Xiaozhong zhitong lotion that are able to reduce swelling， promote ulcer healing and analgesia， and to provide reference for it’s secondary development of ointment preparation. METHODS： Water elution fraction and 20％， 40％， 60％， 95％ ethanol elution parts were separated by D101 macroporous resin from Xiaozhong zhitong lotion. 120 SD rats were randomly divided into normal group （n＝8） and modeling group （n＝112）. Rats in the normal group were not treated. Hemorrhoids model was established in the model group by injecting 75％ glacial acetic acid into the perianal skin to induce perianal ulcer. 96 model rats were randomly divided into model group [blank ointment matrix 0.51 g/（kg·d）]， positive group [Mayinglong ointment， 0.51 g/（kg·d）]， high-dose and low-dose groups of Xiaozhong zhitong lotion and it’s water elution fraction and 20％， 40％， 60％ ethanol elution parts [8.34， 2.78 g/（kg·d） by crude drug， all make into containing drug ointment]， with 8 rats in each group. The periphery of the anus was smeared with relevant medicine， twice a day， for consecutive 7 d. The local symptoms around the anus of rats 3 and 7 days after administration and the pathological morphology of the local mucosa around the anus of rats 7 days after administration were observed and scored respectively. The effects of each elution part for reducing swelling and promoting ulcer healing were investigated. 120 ICR mice were randomly divided into model group [blank ointment matrix 1.03 g/（kg·d）]， positive group [Mayinglong ointment 1.03 g/（kg·d）]， high-dose and low-dose groups of Xiaozhong zhitong lotion and it’s each elution part [16.65， 5.55 g/（kg·d） by crude drug， all make into containing drug ointment]， with 10 mice in each group. Transdermal administration， twice a day， for consecutive 7 d. After 30 min of last administration， the latency time and 15 min writhing times of mice were detected by designing acetic acid writhing test； pain threshold of mice was determined by hot-plate pain test so as to investigate systemic and local analgesic effects of each elution part. RESULTS： In the detumescence and ulcer healing test， compared with normal group， the score of local symptoms around anus at 3rd and 7th day of administration as well as pathological score of hemorrhoids local mucosa were increased significantly in model group （P＜0.01）. Compared with model group， above scores of positive group， 40％ ethanol elution high-dose and low-dose groups were decreased significantly （P＜0.05 or P＜0.01）. The scores of local symptoms around anus in 20％ ethanol elution part high-dose group at 3rd and 7th day after medication as well as 60％ ethanol elution part group at 7th day after medication were decreased significantly （P＜0.05）. In analgesia test， compared with model group， writhing latency time was shortened significantly and 15 min writhing times was decreased significantly in positive group， 40％ ethanol elution part high-dose and low-dose groups as weel as 60％ ethanol elution part high-dose group （P＜0.05 or P＜0.01）； writhing latency time of 20％ ethanol elution part high-dose group was shortened significantly （P＜0.05）. Pain threshold of mice was increased significantly in positive group， 40％ ethanol elution part high-dose and low-dose groups after medication （P＜0.05 or P＜0.01）. CONCLUSIONS： The 40％ ethanol elution part from Xiaozhong zhitong lotion is the effective part that can reduce swelling， promote ulcer healing and analgesia.