In order to explore the scientific connotation of "Fangzhengduiying (formula corresponding to pattern types)", "Qiyinliangxuzheng (Qi and Yin deficiency pattern)" of myocardial ischemia rat model and GC-TOF/MS based metabonomic method were used for comparing the effects of Sheng-mai injection, Salvia injection and propranolol in the present study. After data processing and pattern recognition, Sheng-mai injection showed better efficacy than the other two drugs in accordance with not only visual observation from PLS-DA scores plots but also the number of abnormal endogenous compounds restored to the normal level. Further studies showed that Sheng-mai injection could normalize the level of plasma endothelin-1, the index related to cardiovascular diseases and sleep disorders, which verified the results of metabonomics. Finally, the regulated metabolites and related metabolic pathways were analyzed, and it was supposed that the effects of Sheng-mai injection involved in the alternation of energy metabolism, lipid metabolism, amino acids metabolism, and so on. These findings provided scientific evidence to Shengmai "Fang" used for "Qi and Yin deficiency pattern" correspondingly, indicating that metabonomics has great potential in traditional Chinese medical research, which provides a novel approach and way to modernization of traditional Chinese medicine.

Objective To investigate the additional diagnostic value of 99Tcm-MDP SPECT/CT over conventional SPECT scan in patients with spinal bone malignancy. Methods Fifty-two patients (mean age (56±14) years) with suspicious spinal bone diseases underwent both bone SPECT and SPECT/CT imaging right after 99Tcm-MDP whole-body planar bone scintigraphy. All patients were pathologically diagnosed by either spine operation or trans-spinal biopsy. The images were evaluated by two independent reviewers; inter-reviewer agreement was evaluated using a weighted Kappa score. Each focus of abnormality was recorded using a 4-point diagnostic confidence scale: benign (B), possibly benign (PB), possibly malignant (PM) or malignant (M). Results Accord to the pathological results, 36 patients had bone malignancy (19 with metastatic tumors and 17 with non-metastatic tumors) and 16 had benign lesions. Over SPECT images, two reviewers rated 73.1% (38/52) and 67.3% (35/52) the lesions as the equivocal (PB or PM) respectively. Over SPECT/CT images, they rated only 25.0% (13/52) and 13.5% (7/52) lesions as the equivocal. Inter-reviewer agreement was 63.5% (weighted Kappa score=0.62) for SPECT and 78.9% (weighted Kappa score=0.81) for SPECT/CT. Conclusions Compared with99Tcm-MDP SPECT, 99Tcm-MDP SPECT/CT results in a significant reduction of equivocal diagnoses for the spinal bone malignancy.

Objective To investigate the added diagnostic value of SPECT/CT imaging over routine planar whole-body bone scintigraphy (WBBS) for bone metastases from lung cancer. Methods One hundred and forty-six patients with lung cancer, confirmed by pathological examination, underwent routine 99Tcm-MDP (1110 MBq) WBBS, followed by SPECT/CT over the regions with indeterminate findings on WBBS. Both WBBS and bone SPECT/CT images were interpreted by two experienced nuclear medicine physicians in consensus as the positive, negative or uncertain bone metastases. The final diagnosis was comfirmed by pathology or clinical follow-up. x2 test was used to compare the differences between the two imaging methods. Results Finally, 45 patients were diagnosed as positive bone metastases and the other 101 as negative. The diagnostic sensitivity of bone SPECT/CT for bone metastases from lung cancer was 93.3%(42/45), singnificantly higher than that of WBBS (64.4%, 29/45) (x2 = 19.944, P＜0.05). The diagnostic accuracy of bone SPECT/CT was 89.7% ( 131/146), much higher than that of WBBS (44.5%,65/146) ( x2 = 69. 598,P ＜ 0.05). The uncertain and incorrect diagnostic rates of bone SPECT/CT and WBBS were 10.3% ( 15/146, raging from 5.3% to 15.2% with 95% confidence interval (CI) ) and 55.5% (81/146, raging from 47. 4% to 63.5% with95% CI), respectively. Conclusion BoneSPECT/CT provides incremental diagnostic value over routine WBBS for bone metastases from lung cancer.

Isoproterenol (ISO)-induced myocardial ischemia animal model has been widely applied to the study of myocardial ischemia and evaluation of drug efficacy. Metabolic profiling of endogenous compounds can make a deep insight into biochemical process of the ISO-induced myocardial ischemia rats. Herein, rats were treated with ISO (2 mg x kg(-1)) for 10 days. After the model was established by measuring myocardial histopathology and plasma creatine kinase, GC/TOF-MS was used to determine endogenous metabolites in plasma and cardiac muscle of rats, and pattern recognition was used to process the data. Results showed that the plasma metabolic profiling of ISO-induced myocardial ischemia rats was significantly different from that of the control, and it had the tendency to the normal state after the discontinue of ISO injection. Besides, the cardiac muscle of rats treated with ISO for 10 days and the normal cardiac muscle could also be separated clearly. The potential biomarkers in plasma and cardiac muscle of model rats had homogeneity and their own specialty. Biochemical metabolic pathway analysis indicated that this myocardial ischemia model was involved in the alternation of energy metabolism, saccharometabolism, lipid metabolism, nucleoside metabolism and amino acid metabolism, and in relationship with oxidative stress. These findings revealed that metabonomics may be a promising tool to evaluate myocardial ischemia rat model induced by ISO and could further extend the study of pharmacodynamic action of drugs at the molecular level.
Animals ; Creatine Kinase ; blood ; Energy Metabolism ; Isoproterenol ; Lipid Metabolism ; Male ; Metabolome ; Metabolomics ; methods ; Myocardial Ischemia ; blood ; chemically induced ; metabolism ; Myocardium ; metabolism ; Oxidative Stress ; Random Allocation ; Rats ; Rats, Sprague-Dawley

In order to explore the scientific connotation of "Fangzhengduiying (formula corresponding to pattern types)", "Qiyinliangxuzheng (Qi and Yin deficiency pattern)" of myocardial ischemia rat model and GC-TOF/MS based metabonomic method were used for comparing the effects of Sheng-mai injection, Salvia injection and propranolol in the present study. After data processing and pattern recognition, Sheng-mai injection showed better efficacy than the other two drugs in accordance with not only visual observation from PLS-DA scores plots but also the number of abnormal endogenous compounds restored to the normal level. Further studies showed that Sheng-mai injection could normalize the level of plasma endothelin-1, the index related to cardiovascular diseases and sleep disorders, which verified the results of metabonomics. Finally, the regulated metabolites and related metabolic pathways were analyzed, and it was supposed that the effects of Sheng-mai injection involved in the alternation of energy metabolism, lipid metabolism, amino acids metabolism, and so on. These findings provided scientific evidence to Shengmai "Fang" used for "Qi and Yin deficiency pattern" correspondingly, indicating that metabonomics has great potential in traditional Chinese medical research, which provides a novel approach and way to modernization of traditional Chinese medicine.
Animals ; Anti-Arrhythmia Agents ; pharmacology ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Endothelin-1 ; blood ; Gas Chromatography-Mass Spectrometry ; methods ; Injections ; Male ; Medicine, Chinese Traditional ; Metabolomics ; methods ; Myocardial Ischemia ; blood ; metabolism ; pathology ; Panax ; chemistry ; Plants, Medicinal ; chemistry ; Propranolol ; pharmacology ; Qi ; Rats ; Rats, Sprague-Dawley ; Salvia ; chemistry ; Schisandraceae ; chemistry ; Yin Deficiency ; metabolism

The resolution of single molecule localization imaging techniques largely depends on the precision of localization algorithms. However, the commonly used Gaussian function is not appropriate for anisotropic dipoles because it is not the true point spread function. We derived the theoretical point spread function of tilted dipoles with restricted mobility and developed an algorithm based on an artificial neural network for estimating the localization, orientation and mobility of individual dipoles. Compared with fitting-based methods, our algorithm demonstrated ultrafast speed and higher accuracy, reduced sensitivity to defocusing, strong robustness and adaptability, making it an optimal choice for both two-dimensional and three-dimensional super-resolution imaging analysis.
Alcohol Oxidoreductases ; analysis ; genetics ; metabolism ; Algorithms ; Animals ; COS Cells ; Cercopithecus aethiops ; Cytochrome P-450 Enzyme System ; analysis ; genetics ; metabolism ; HeLa Cells ; Humans ; Imaging, Three-Dimensional ; Microscopy, Fluorescence ; Normal Distribution ; Plasmids ; metabolism

Objective: To establish human cancer cell strains with stable Cas9 expression, and to validate the gene editing activity of Cas9 for simple gene editing in future study. Methods: Fifteen cancer cell lines of different tissue origins were infected with pLv-EF1α-Cas9-Flag-Neo or pLv-EF1α-Cas9-Flag-Puro by lentivirus and clone selection was employed to screen Cas9 stably expressed cancer cell lines. Afterward designed guide RNA vectors targeting TSC22 gene were transiently transfected into 3 of cell lines, and subsequently the gene editing activity of Cas9 was evaluated by genomic PCR, sequencing and Western blot. Results: Sixty-nine human cancer cell strains with stable Cas9 expression from different cancers were established, and by transient transfection with designed guide RNA, long fragment deletion was detected in TSC22 gene. Conclusions Sixty-nine human cancer cell strains are successfully established with stable expression of Cas9 protein and gene editing activity. These cell strains may be employed in large-scale drug screening, screening of new drug targets and gene function investigation.

The global metabolite profiles of endogenous compounds of Wistar rats from 12 to 20 weeks old were investigated to take deep insight into and get better understanding of the pathogenesis of development and aging. Plasma from Wistar rats at 12, 14, 16, 18, and 20 weeks old were analyzed using GC/TOFMS. Multivariate data analysis was then used to process the metabonomic data which indicated excellent separation between different weeks and showed that the metabolic profiles of the samples changed with age, enabling age-related metabolic trajectories to be visualized. Decreased concentrations of citric acid, cis-aconitic acid, 9-(z)-hexadecenoic acid along with increased levels of hexanedioic acid, alpha-tocopherol, 3-indole propionic acid, etc contributed to the separation. Several major metabolic pathways were identified to be involved in metabolic regulation. This suggests that GC/TOFMS-based metabonomics is a powerful alternative approach to identifying potential biomarkers and investigating the physiological developments of aging and it is important to employ suitable age-match control group in metabonomic study of physiological monitoring, drug safety assessment, and disease diagnosis, etc.
Aconitic Acid ; blood ; Adipates ; blood ; Aging ; blood ; physiology ; Animals ; Biomarkers ; blood ; Chromatography, Gas ; methods ; Citric Acid ; blood ; Indoles ; blood ; Male ; Metabolome ; Metabolomics ; Multivariate Analysis ; Palmitic Acids ; blood ; Propionates ; blood ; Rats ; Rats, Wistar ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods ; alpha-Tocopherol ; blood

ObjectiveTo investigate the effects of cynomorium songaricum (CS) decoction on the testis weight, serum testosterone level, and sperm parameters of rats with oligoasthenospermia (OAS), explore its action mechanism of improving the proliferation of undifferentiated spermatogonial cells, and provide some experimental and theoretical evidence for the development of new Chinese drugs for OAS.

METHODSThirty 8-week-old male SD rats were randomly divided into five groups of equal number: blank control, model control, high-dose CS, medium-dose CS, and low-dose CS. OAS models were established by intraperitoneal injection of cyclophosphamide and, a month later, treated intragastrically with normal saline or CS at 2, 1, and 0.5 g per kg of the body weight per day, all for 4 weeks. Then, the testes of the animals were harvested to obtain the testicular weight, sperm concentration and motility, and the level of serum testosterone (T), detect the expressions of the transcription factor 1 (Oct4), Thy-1 cell surface antigen (Thy1), promyelocytic leukemia zinc finger (PLZF), KIT proto-oncogene receptor tyrosine kinase (C-kit) and glial cell-derived neurotrophic factor (GDNF) in the testis tissue of the rats in the low-dose CS group by real-time PCR.

RESULTSThe testis weights in the blank control, model control, high-dose CS, medium-dose CS, and low-dose CS groups were (1.52±0.06), (1.55±0.06), (1.43±0.30), (1.35±0.40) and (1.34±0.04) g, respectively, not significantly different in the blank and model controls from those in the CS groups (P>0.05). The visual field sperm count per 10 HP was significantly increased in the high-, medium-, and low-dose CS groups (202±20, 196±5 and 216±25) as compared with the blank and model controls (200±15 and 134±30) (P<0.05). The mRNA expressions of the Oct4, Thy1, PLZF and GDNF genes were remarkably higher in the low-dose CS group than in the controls (P<0.05), but that of the C-kit gene showed no significant difference from the latter (P>0.05). The visual field sperm motility per 10 HP was markedly increased in the blank control (［52.1±5.5］%), model control (［38.1±2.5］%), high-dose CS (［59.1±9.5］%), medium-dose CS (［58.7±9.5］%), and low-dose CS (［49.6±1.0］%) groups, and so was the level of serum testosterone (［190±87.5］, ［82.5±25.8］, ［229±75.6］, ［331±86.7］ and ［185±82.4］ mmol/L), both remarkably higher in the CS groups than in the model controls (P<0.05) but with no statistically significant difference between the CS groups and the blank controls (P>0.05).

CONCLUSIONSCS can significantly improve sperm concentration, sperm motility and serum T level in OAS rats, probably by inducing the expression of GDNF in the rat Sertoli cells, promoting the proliferation of undifferentiated spermatogonial cells, and enhancing spermatogenesis.

OBJECTIVE: To explore clinical effect of arthroscopy-assisted rotator cuff tendon transfer in treating irreparable rotator cuff tears (IRCT). METHODS: From May 2015 to May 2018, 23 patients with unrepairable rotator cuff tears were treated with arthroscopy-assisted rotator cuff tendon transfer, and 21 patients were followed up finally, including 8 males and 13 females, aged from 48 to 82 years old with an average of(64.3±9.1) years old;the courses of disease ranged from 6 to 36 months with an average of (14.0±6.4) months. American Rotator and Elbow Surgeons Score(ASES) and Constant-Murley score were used to evaluate clinical efficacy before surgery and at the latest follow-up. RESULTS: All 21 patients were followed up for 36 to 54 months with an average of (39.4±4.4) months. Axillary incision of 1 patient was redness, swelling and exudation after surgery, which healed after 3 weeks of dressing change, and exudate culture was negative. At the latest follow-up, MRI showed partial tearing of the metastatic tendon in 2 patients, but pain and movement of the affected shoulder were still better than before surgery. ASES increased from preoperative (41.0±9.6) scores to the latest follow-up (75.6±14.0) scores, and had statistical difference (t=10.50, P<0.01). Constant-Murley score increased from (49.8±7.1) scores before operation to (67.5±11.6) scores at the latest follow-up (t=11.27, P<0.01). CONCLUSION Arthroscopic assisted latissimus dorsalis tendon transposition restores physiological and anatomical structure of glenohumeral joint by reconstructing balance of horizontal and vertical couples of shoulder joint, thus achieving the stability of the shoulder joint, relieving shoulder pain and improving shoulder joint function.
Male ; Female ; Humans ; Middle Aged ; Aged ; Aged, 80 and over ; Rotator Cuff Injuries/surgery* ; Superficial Back Muscles ; Rotator Cuff ; Treatment Outcome ; Shoulder Joint/surgery* ; Tendon Transfer ; Arthroscopy ; Range of Motion, Articular/physiology*