To establish a method for simultaneous determination of dehydrotumulosic acid, polyporenic acid C, 3-epi-dehydrotumulosic acid, dehydropachymic acid and pachymic acid in Poria, a RP-HPLC method detected by UV wavelengths switch had been developed, including 210 nm (48-55 min) for pachymic acid and 241 nm (0-48 min) for dehydrotumulosic acid, polyporenic acid C, 3-epi-dehydrotumulosic acid, dehydropachymic acid, separately. The system consisting of a Kromasil C18 column (250 mm x 4.6 mm, 5 microm) and a mixture of acetonitrile and 0.05% phosphate acid as the mobile phase was adopted; The flow rate was 1.0 mL x min(-1). The linear response range was 30.5-610.0 microg x mL(-1) (r = 0.999 6) for dehydrotumulosic acid, 12.66-253.2 microg x mL(-1) (r = 0.999 5) for polyporenic acid C, 2.99-59.7 microg x mL(-1) (r = 0.999 7) for 3-epi-dehydrotumulosic acid, 6.13-122.5 microg x mL(-1) (r = 0.999 5) for dehydropachymic acid and 11.3-226.0 microg x mL(-1) (r = 0.9995) for pachymic acid. The average recoveries of these compounds were 98.5% (RSD = 1.9%), 99.4% (RSD = 1.7%), 97.9% (RSD = 1.2%), 96.7% (RSD = 2.5%) and 97.9% (RSD = 2.3%), respectively. The method is simple, accurate and reproducible for quality control of Poria.

OBJECTIVE: To compare the effectiveness of sequential treatment for subjective and objective benign paroxysmal positional vertigo(BPPV). METHOD: The efficacy of sequential treatment for nineteen vertical semicircular canal S-BPPV patients(Group A) and forty-five vertical semicircular canal O-BPPV patients(Group B) who were admitted to our hospital between January 2014 and July 2015 was retrospectively analyzed and compared, treatment of five cases with horizontal semicircular canal S-BPPV was reported here as well. RESULT: The number of repositioning maneuver for Group A was 2 - 6 times (average: 4. 21±1. 18) while 2 - 3 times (average: 2. 29 ± 0. 46) for Group B. There was significant difference between the two groups(P<0. 01). The effective rate of initial treatment for Group A was 78. 9%(15/19) while 82. 2%(37/45)for Group B. After one weeks treatment, the effective rate for Group A changed to be 89. 5% (17/19)while 88. 9% (40/45) for Group B. Non-significant difference was found in the two groups neither in the initial nor one weeks treatment. On three months' follow-up, one in Group A and two in Group B relapsed, and they were cured after treated in the initial treatment. The recurrence for Group A was 5. 3%(1/19) while 4. 4%(2/45) for Group B and there was non-significant difference. The five cases with horizontal semicircular canal S-BPPV were completely free of disease after undergoing treatment. CONCLUSION The sequential treatment is effective for both vertical semicircular canal S-BPPV and vertical semicircular canal O-BPPV, but the former needs more repositioning maneuver times. The repositioning maneuver, combined with drug therapy and head-shake-in-supine-position method is recommended for horizontal semicircular canal S-BPPV.
Benign Paroxysmal Positional Vertigo ; therapy ; Humans ; Patient Positioning ; Recurrence ; Retrospective Studies ; Semicircular Canals ; physiopathology ; Treatment Outcome

OBJECTIVETo explore the expression of p-p38 MAPK protein and the number of astrocytes expressing p-p38 MAPK in CA1 hippocampus in rats during the induction of brain ischemic tolerance induced by intermittent hypobaric hypoxia (IH) preconditioning.

METHODSThirty healthy adult male Wistar rats were randomly divided into 6 groups (n = 5 in each group): sham 0 min group, IH + sham 0 min group, sham 7 d group, IH + sham 7 d group, Ischemia (Is) 7 d group, and IH + Is 7 d group. Neuropathological evaluation was performed by thionine staining in CA1 hippocampus in rats. The expression of p-p38 MAPK in CA1 hippocampus was observed by immunohistochemical staining. And the number of astrocytes expressing p-p38 MAPK was observed by immunofluorescent double labeling.

RESULTSThe results showed that IH preconditioning induced brain ischemic tolerance successfully. At the same time, IH preconditioning obviously up-regulated the expression of p-p38 MAPK protein in CA1 hippocampus, and also increased the number of astrocytes expressing p-p38 MAPK.

CONCLUSIONIt might be concluded that IH preconditioning induced brain ischemic tolerance by up-regulating the expression of p-p38 MAPK protein in pyramidal neurones and astrocytes.

Animals ; Astrocytes ; enzymology ; pathology ; Brain Ischemia ; enzymology ; pathology ; Disease Models, Animal ; Hippocampus ; enzymology ; Hypoxia ; Ischemic Preconditioning ; methods ; Male ; Phosphorylation ; Pressure ; Rats ; Rats, Wistar ; p38 Mitogen-Activated Protein Kinases ; metabolism

Objective To conclude and compare the technical characteristics and clinical efficacy of retroperitoneal and transperitoneal approach for robotic partial nephrectomy.Methods 418 patients received robotic partial nephrectomy conducted by one surgeon in the urology department of PLA General Hospital from 2013, December to 2015, October, including 157 cases in retroperitoneal approach and 261 cases in transperitoneal approach.There were no significant difference between the transperitoeal and retroperitoneal approach in patient age(51 years, 51 years, P=0.593), BMI(25.5 kg/m2,25.6 kg/m2, P=0.331), gender(male/female:111/46,186/75,P=0.184), location(left/right,80/77,123/138,P=0.575), tumor size(3.1cm,3.5cm,P =0.356), comorbidities, proportion of solitary kidney(11.5%, 9.6%,P=0.253) or bilateral tumors(1.3%,3.4%,P=0.179), RENAL score(6(5-8),6(5-8),P=0.083), ASA score(2 vs.2)(P=0.310) and preoperative serum creatinine(74.7 μmol/L,76.7 μmol/L, P=0.736).Patients in the retroperitoeal approach group were more likely to have abdomen surgery history (21.7%,14.2%,P=0.049).there was significant difference between two approaches in tumor location in the kidney ( anterior/posterior/others: 9/140/8, 212/36/13, P <0.001 ) Results The operation time (105min, 115min, P =0.041 ) and warm ischemia time ( 15min, 20min, P <0.001 ) were shorter and estimated blood loss(50ml,75ml,P<0.001) was less in the retroperitoneal group.No significant difference was observed in postoperative 24h blood loss(45ml,50ml,P =0.093), intraoperative transfusion rate (2.5%,6.5%,P=0.072) and collective system injury(24.8%,27.6%,P =0.539) between the two groups.Peritoneum break occurred in 25 cases in retroperitoneal group, which were repaired by Hem-o-lock.One intestinal injury occurred in transperitoneal group and was repaired intraoperatively.Three and nine cases in the retroperitoneal and transperitoneal group were converted to radical nephrectomy.One case in the transperitoneal group was found positive margin and received laparoscopic radical nephrectomy.Two liver injury and five spleen injury occurred in the transperitoneal group.For postoperative food intake(1 day, 2 days, P <0.001 ) , the length of indwelling drainage ( 3 days, 4 days, P <0.001 ) and the length of postoperative stay ( 5 days, 6 days, P =0.001 ) , the retroperitoneal group had one day shorter than the transperitoneal group.With the median following-up time of both group, 11 months (IQR:6-16) in RPRPN group and 12 months ( IQR: 7-19 ) , no subject with a tumor recovery or metastasis.Conclusions Retroperitoneal robotic partial nephrectomy is feasible and effective, which is similar to transperitoneal approach.The retroperitoneal approach is superior to transperitoneal approach for posterior, lateral and superior renal tumors.

Acupuncture has remarkable effects of pain relieving and functional restoration on injuries of soft tissue and joint due to military training. As more and more attention has been attached to the impact of psychological states and biorhythm disorder on the fighting ability of military staff, acupuncture has found its place in treating chronic fatigue, combat stress reaction, traumatic brain injury and post-traumatic stress disorder as well as regulating circadian rhythms. The therapeutic effect of acupuncture in military training-related physical damage and psychological trauma has already been proved by numerous clinical practices and researches. It is held that using acupuncture as an alternative could not only save medical resources, but also enhance the fighting ability of the army. However, the current clinical studies is facing the problem of limited sample size. Therefore, randomized controlled trials in large scale and multiple centers should be further carried out toward military staff, so as to provide more speaking evidences to the prevention and treatment of physical and psychological diseases.
Acupuncture Therapy ; Adult ; Female ; Humans ; Male ; Middle Aged ; Military Personnel ; psychology ; Pain Management ; Stress Disorders, Post-Traumatic ; therapy ; Wounds and Injuries ; therapy ; Young Adult

A stable and efficient L-Malic acid accumulation mutant strain Rhizopus oryzae ME-M15 was discovered occasionally in the mutation breading for fumaric acid producers. Rhizopus oryzae ME-M15 gave a L-Malic acid output of 16.3 g/L on average after fermentation for 96 hours, more than 3 times than that of the parent strain ME-F10. In addition, other metabolites such as ethanol and fumaric acid were re-markably decreased in accordance with the depressed activity of the cytosolic isoenzyme of fumarase and alcohol dehydrogenase in strain ME-M15, while the activity of the pyruvate carboxylase had no significant difference.

The bioconversion of lignocellulosic materials for organic acid had great meaning to both economic and social benefit,owning to its renewable character,abundance and low price.In order to find out the key to the efficient utilization of lignocellulose,kinds of fermentation strategies to product organic acid(such as loctic acid,funaric acid)from lignocellulcsic were discussed,also the xylose metabolic pathway of Rhizopus sp.was extensively reviewed,and it find out that the low conversion of xylose was the bottle-block.

Purpose Sustained attention dysfunction is a common symptom of patient with attention-deficit/hyperactivity disorder (ADHD).To reveal the neural mechanism of the abnormality of sustained attention of patients with ADHD,the cerebral blood flow (CBF) abnormalities in patients were studied by perfusion imaging.Materials and Methods Sixteen children with ADHD and twenty normal controls treated at the 401st Hospital of PLA from June 2013 to September 2015 underwent 3D arterial spin labeling (3D-ASL)scanning with GE 3.0T MRI scanner.The participants were performed four continuous sections of sustained attention to response task (SART) for 20 minutes in the scanner.Using SPM 8 toolkit,the local CBF values of both groups were compared in a voxel-wise manner,and their correlations with response time and target accuracy of SART were analyzed.Results When performing the SART,the patients with ADHD showed significantly inhibited trend of increasing CBF in the anterior cortex like dorsal cortex,medial prefrontal cortex,and motor area;however,they presented enhanced trend of increasing CBF in the posterior cortex such as posterior cingulate cortex and parietooccipital sulcus (P＜0.01);the change of CBF in the dorsal prefrontal cortex and that in the precentral and postcentral gyrus had significant correlation with response time of SART task and targeting ratio (dorsal prefrontal cortex:r=0.745,P＜0.001;r=0.591,P＜0.001;r=-0.521,P＜0.001.Precentral and postcentral gyrus:r=0.579,P＜0.001).Conclusion Patients with ADHD show different CBF redistribution between anterior and posterior cerebral cortex in performing SART,and the abnormal CBF shows significant correlations with behavioral metrics,which reflects the mechanism of sustained attention dysfunction of patients with ADHD.

To establish a method for simultaneous determination of dehydrotumulosic acid, polyporenic acid C, 3-epi-dehydrotumulosic acid, dehydropachymic acid and pachymic acid in Poria, a RP-HPLC method detected by UV wavelengths switch had been developed, including 210 nm (48-55 min) for pachymic acid and 241 nm (0-48 min) for dehydrotumulosic acid, polyporenic acid C, 3-epi-dehydrotumulosic acid, dehydropachymic acid, separately. The system consisting of a Kromasil C18 column (250 mm x 4.6 mm, 5 microm) and a mixture of acetonitrile and 0.05% phosphate acid as the mobile phase was adopted; The flow rate was 1.0 mL x min(-1). The linear response range was 30.5-610.0 microg x mL(-1) (r = 0.999 6) for dehydrotumulosic acid, 12.66-253.2 microg x mL(-1) (r = 0.999 5) for polyporenic acid C, 2.99-59.7 microg x mL(-1) (r = 0.999 7) for 3-epi-dehydrotumulosic acid, 6.13-122.5 microg x mL(-1) (r = 0.999 5) for dehydropachymic acid and 11.3-226.0 microg x mL(-1) (r = 0.9995) for pachymic acid. The average recoveries of these compounds were 98.5% (RSD = 1.9%), 99.4% (RSD = 1.7%), 97.9% (RSD = 1.2%), 96.7% (RSD = 2.5%) and 97.9% (RSD = 2.3%), respectively. The method is simple, accurate and reproducible for quality control of Poria.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Lanosterol ; analogs & derivatives ; analysis ; Plants, Medicinal ; chemistry ; Poria ; chemistry ; Quality Control ; Reproducibility of Results ; Spectrophotometry, Ultraviolet ; methods ; Triterpenes ; analysis

To purify the recombinant human BMP-6 protein and to establish its in vitro bioassay method. The cDNA encoding the mature peptide of hBMP-6 protein was amplified by reverse transcription-polymerase chain reaction (RT-PCR), using human placental mRNA as template, and subcloned into the high-expression vector pET-15b under the control of T7 lac promoter. The resulting construct, pET-BMP6, was then transformed into an Escherichia coli strain BL21 (DE3) for the production of recombinant hBMP-6 protein (rhBMP-6). After 4 hours of induction by isopropyl-beta-D-thiogalactoside (IPTG), rhBMP-6 (approximately 15kD) was expressed and formed inclusion bodies, contributing up to 10% of the total bacterial protein. The inclusion bodies were isolated and redissolved in 8mol/L urea, and the denatured rhBMP-6 was purified to 95% purity by CM-Cellulose 32 ion exchange chromatography (IEC). The osteoinductivity of rhBMP-6 was measured by the expression of some of the osteoblast differentiation marker genes in rhBMP-6-treated C3H10T1/2 cells as reflected by determinations of alkaline phosphatase (ALPase) activity and semi-quantitative RT-PCR. At the end of the purification process, about 80% of rhBMP-6 formed disulphide-linked homodimers after refolding during renaturation. The apparent size of the protein was 30kD on non-reducing SDS-PAGE, similar to that of the native form of hBMP-6. The enzyme assays showed that the ALPase activity was increased in a dose-dependent manner with the treatment of rhBMP-6. After the addition of 300ng/mL of rhBMP-6, the ALPase activity of C3H10T1/2 cells increased significantly. The activity of rhBMP-6 used was comparable to about 70% of that of the standard hBMP-6 derived from eukaryotic cells. RNA extraction data also showed rhBMP-6 stimulated expression of osteoblast marker genes, including type I collagen, osterix, and osteocalcin in a time-dependent manner. After 5 days of treatment, their level of expression was increased to 3 times that of controls. Bone morphogenetic protein (BMP)-6, a member of the 60A subgroup of the bone morphogenetic protein (BMPs) family, plays a pivotal role in bone formation. Previous evidence showed that BMP-6 is selectively up-regulated by estrogen, suggesting its potential role in the treatment of osteoporotic fractures, especially for menopausal osteoporosis. Our present study demonstrates that the recombinant hBMP-6 produced in Escherichia coli is able to induce pre-osteoblastic cells to differentiate into osteoblasts in vitro, and analysis of mRNA expression of type I collagen, osterix, and osteocalcin can be also a method for measuring the osteoinductivity of BMP. This provides the basis for further studies on ectopic bone formation in the body and for the development of auxiliary drugs for the treatment of osteoporotic fractures.
Alkaline Phosphatase ; metabolism ; Animals ; Biological Assay ; methods ; Bone Morphogenetic Protein 6 ; genetics ; isolation & purification ; metabolism ; pharmacology ; Cell Line ; Chromatography, Ion Exchange ; Electrophoresis, Polyacrylamide Gel ; Enzyme Activation ; drug effects ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; drug effects ; genetics ; Humans ; Isopropyl Thiogalactoside ; pharmacology ; Mice ; Plasmids ; Reverse Transcriptase Polymerase Chain Reaction