Moving cup of cupping therapy is studied in the present paper systematically. According to the experiences of clinical practice, duration, speed and intensity of moving cup has been quantified. Fifteen basic manipulations have been evolved, and 10 protocols for clinical application recommended. "Moving cup according to 3 factors of duration, speed and intensity" has been advanced on the basis of acupuncture therapy. And it has been concluded that the effects of warming needling, simple cupping, scraping, massage and medication can be achieved by moving cup with different techniques and clinical protocols.
Humans ; Medicine, Chinese Traditional ; adverse effects ; methods ; Therapeutics ; adverse effects ; methods

OBJECTIVE The present study was aimed to investigate the role of Wnt/β-catenin sig-naling in spinal VGLUT2 regulation and neuropathic pain. METHODS To elucidate the association be-tween VGLUT2 and neuropathic pain,we determined the expression and distribution characteristics of VGLUT2 in mice subjected to spared nerve injury(SNI),and then observed the effects of two VGLUT2 targeting shRNAs on mechanical allodynia and glutamate release.The effects of Wnt/β-catenin signal-ing on VGLUT2 expression and pain behavior were investigated by using Wnt agonist,Wnt1,and Wnt/β-catenin pathway inhibitor XAV939 in SNI mice.RESULTS SNI surgery induced significant up-regula-tion of VGLUT2 on postoperative days 7,14,and 21.Double immunofluorescence labeling of VGLUT2 with NeuN,MAP2,Iba-1,or GFAP showed that VGLUT2 was mainly expressed in neurons in the dor-sal horn of the spinal cord after SNI(NeuN,MAP2).Intrathecal administration of VGLUT2 shRNAs be-fore or after SNI surgery significantly decreased mechanical allodynia and glutamate release. Mean-while,Wnt1/β-catenin signaling increased significantly after SNI surgery.Over-expression of β-catenin in PC12 cells increased VGLUT2 protein level,intrathecal administration of Wnt agonist or Wnt1 signifi-cantly increased VGLUT2 protein expression in spinal cord, while Wnt/β-catenin pathway inhibitor XAV939 decreased VGLUT2 expression in PC12 cells and spinal cord.Additionally,intrathecal admin-istration of XAV939 7 days after SNI significantly attenuated mechanical allodynia in mice, which was in accordance with down-regulation of VGLUT2 protein levels.VGLUT2 shRNAs significantly attenuat-ed Wnt agonist or Wnt1 induced mechanical allodynia. CONCLUSION Wnt1/β-catenin signaling path-way up-regu-lates the spinal VGLUT2 expression,and this regulation is involved in neuropathic pain behavior.

Lysine decarboxylase is a key enzyme involved in the upstream biosynthesis of lycopodium alkaloids (LAs) such as huperzine A, contributing to the decarboxylation of lysine to 1,5-pentanediamine (cadaverine). Three lysine decarboxylase genes (HsLDC-L1, HsLDC-L2, HsLDC-L3) were successfully cloned from Huperzia serrata using transcriptomic sequence data mining strategy combined with reverse transcription PCR. The physicochemical properties, secondary and tertiary structures, amino acid identities, and evolutionary relationship of the three LDCs were analyzed by online bioinformatics analysis platforms and DNAMAN, MEGA 7.0 software, revealing that all of these proteins had the conserved PLP binding domain and active site residues were completely conserved in LDCs. Phylogenetic analysis showed that these LDCs were located in the same branch as other known LDCs from LA-producing plants. Accordingly, the ORFs of these three HsLDCs were inserted into different expression plasmids for further expression in E. coli. However, only HsLDC-L1 was successfully expressed in E. coli BL21 (DE3) by inserting into a pCold TF vector. The recombinant protein was purified by Ni²⁺ affinity chromatography purification. HsLDC-L1 contains 469 amino acid residues, with a calculated molecular weight of 50.50 kDa. HsLDC-L1 expectedly catalyzed the decarboxylation of lysine to produce cadaverine. In addition, HsLDC-L1 can also catalyze the generation of putrescine from ornithine. However, it cannot catalyze the decarboxylation of tyrosine, phenylalanine, tryptophan and histidine. The results not only provide insight into the biosynthesis of LAs including huperzine A, but also provide a critical genetic element for the overproduction of Δ¹-piperideine and pelletierine, the essential biosynthetic precursors of LAs, using synthetic biology strategies.

OBJECTIVETo investigate risk factors associated with peritoneal metastasis in colorectal cancer.

METHODSClinicopathological data of 873 patients with colorectal cancer treated at the Nanfang Hospital of Southern Medical University from January 2003 to December 2008 were retrospectively analyzed. The associations between peritoneal metastasis and clinicopathological factors were evaluated by univariate analysis including independent-samples t test and χ(2) test, and multivariable logistic regression analysis.

RESULTSUnivariate analysis showed that age(P=0.032), tumor size(P=0.001), tumor differentiation(P=0.008), depth of bowel wall invasion(P=0.000), lymph node metastasis(P=0.000), serum carcinoembryonic antigen (CEA) level(P=0.003), and serum carbohydrate antigen 19-9(CA19-9) level(P=0.009) were associated with peritoneal metastasis. On multivariable logistic regression analysis, depth of bowel wall invasion, lymph node metastasis, serum CEA level and CA19-9 level remained as significant risk factors(all P<0.05).

CONCLUSIONDepth of bowel wall invasion, lymph node metastasis, serum CEA level, and CA19-9 level are risk factors for peritoneal metastasis in colorectal cancer.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Chi-Square Distribution ; Colorectal Neoplasms ; diagnosis ; pathology ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Neoplasm Metastasis ; Peritoneum ; pathology ; Prognosis ; Retrospective Studies ; Risk Factors ; Young Adult

OBJECTIVETo construct a clinical database for laparoscopic colorectal surgery in Chinese population using computerized technique.

METHODSA clinical database system was constructed and used in multi-/single-center studies on laparoscopic colorectal surgery.

RESULTSThe data of more than 1200 cases in the Nanfang Hospital during the past year were collected retrospectively. The database was used as the platform for "Southern China Laparoscopic Colorectal Surgery Study Group (SCLCSG)" and was used in the first stage of the clinical research of "Multicenter retrospective study of laparoscopic and open procedure for colorectal cancer" among 11 hospitals in Southern China. In order to test the system, the database was also used in "comparative study on oncologic results of laparoscopic versus open radical resection for rectal carcinoma".

CONCLUSIONSThe evaluation system is reliable and efficient. This system has established a clinical database for laparoscopic colorectal cancer surgery and can be widely applied for the clinical research for colorectal cancer.

Colectomy ; Colorectal Neoplasms ; surgery ; Colorectal Surgery ; Data Mining ; Databases, Factual ; Humans ; Laparoscopy ; Treatment Outcome

OBJECTIVETo observe effects of Fengbaisan (, FBS) on the expression of matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in lung tissue of rats with chronic obstructive pulmonary disease (COPD) and to investigate the preventive and therapeutic mechanisms of FBS.

METHODSThe COPD rat model was established by cigarette smoke exposure and lipopolysaccharide (LPS) intra-tracheal dripping. The histopathological changes of lung tissue was observed via hematoxylin/eosin staining. The expression of MMP-9 and TIMP-1 in lung tissue was measured by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry.

RESULTSThe typical histopathological changes of COPD were displayed in the model group, Ambroxol Hydrochloride group and FBS group, and the pathological lesions in the FBS group were less than those in the model group. The expression of MMP-9 and TIMP-1 in the model group increased significantly compared with those in the normal group (P<0.05). After treatment for successive 28 days, the expression of MMP-9 and TIMP-1 in the FBS group decreased remarkably as compared with the model group (P<0.05).

CONCLUSIONSFBS can regulate MMP-9/TIMP-1 imbalance to prevent airway and lung parenchyma remodeling process via reducing the expression of MMP-9 and TIMP-1 in the lung tissue of COPD rats, and this may be a possible therapeutic mechanism of FBS on COPD.

Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Gene Expression Regulation, Enzymologic ; drug effects ; Immunohistochemistry ; Lung ; drug effects ; enzymology ; pathology ; Male ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; enzymology ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Tissue Inhibitor of Metalloproteinase-1 ; genetics ; metabolism

The treatment of fracture has been greatly improved, but the incidence of nonunion is still high and which is a challenge that orthopedic clinicians. The treatment of nonunion has been the concern to the scholars. New technologys of surgical and non-surgical therapies continue to emerge, and achieve good clinical efficacy. In particular the development of non-surgical therapy has brought hope for non-invasive treatment of nonunion. This paper attempts to make a review of the status and progress of non-surgical treatment of bone nonunion which are more commonly used in clinical.
Bone Marrow Transplantation ; Bone Morphogenetic Proteins ; therapeutic use ; Fractures, Ununited ; therapy ; Genetic Therapy ; High-Energy Shock Waves ; therapeutic use ; Humans ; Transplantation, Autologous ; Ultrasonic Therapy

Based on the deletion information of high virulent PRRSV genome, 3 oligonucleotide primer were designed and synthesized. Specific and sensitive reverse transcription-PCR (RT-PCR) assays were de-veloped for the detection of high virulent PRRSV. The sensitivity and specificity of RT-PCR assays were evaluated, the results showing that the detection limit of the assay was found to be 0.265 pg of tissue total RNA, and the protocol have no cross-reaction with classical swine fever virus, porcine circovirus type 2,pseudorabies virus, streptococcus, haemophilus parasuis and Escherichia coli. Then 36 cell cultures, two PRRSV live vaccine strains and 184 clinical specimens from 52 farms were tested. Five PRRSV field iso-lates were the high virulent PRRSV; two PRRSV live vaccine strains from normal PRRSV, and 123 speci-mens from 42 farmer were positive (only 1 specimen was normal PRRSV). This RT-PCR method proved to be accurate differential diagnosis of the high virulent PRRSV and normal PRRSV with the characteristics of rapidity, sensitivity and specificity, and has a strong clinical relevance.

OBJECTIVETo establish a NOD/SCID mouse model with human immune reconstitution and observe its immune response to human triple-negative breast cancer xenograft.

METHODSTwenty-four NOD/SCID mice without immune leakage were subjected to cyclophosphamide (CTX) treatment 3 days prior to immune reconstitution with human peripheral blood mononuclear cell (PBMC) injection and subcutaneous transplantation of human triple-negative breast cancer MDA-MB-231 cells, CTX treatment and PBMC injection without tumor cell transplantation, MDA-MB-231 cell transplantation only, or no treatments. The tumor growth and immune responses of the mice were observed at regular intervals.

RESULTSCompared with the tumor-bearing mice, the tumor-bearing mice with immune reconstitution showed prolonged incubation period of tumor formation, slower tumor growth rate and increased survival rate. Human IgG and CD3(+) T cells were detected in the peripheral blood of the mice 1 week after human PBMC injection. The percentage of CD3(+) T cells in the spleen cells was 55.3% at 9 weeks in tumor-bearing mice with immune reconstitution and 52.7% in tumor-bearing mice without immune reconstitution. The spleen index of the tumor-bearing mice with immune reconstitution was much higher than that in mice with only immune reconstitution and the control mice (9.64 vs 3.82∓0.31 and 1.51∓0.14 mg/g).

CONCLUSIONA stable NOD/SCID mouse model with immune reconstitution has been established successfully, which shows immune responses to triple-negative breast cancer xenografts and allows studies of immunological therapy study of triple-negative breast cancer.

Animals ; Disease Models, Animal ; Humans ; Leukocytes, Mononuclear ; Mice ; Mice, Inbred NOD ; Mice, SCID ; Neoplasm Transplantation ; Spleen ; immunology ; Triple Negative Breast Neoplasms ; immunology

OBJECTIVETo observe the function of wnt/β-catenin signal pathway on the process that epimedium-derived flavonoids (EFs) regulate the balance between osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats, and to provide an experimental evidence for the mechanism of EFs on treating postmenopausal osteoporosis.

METHODSBone marrow stromal cells from ovariectomized rats were separated and cultivated in the condition of osteoinductive medium or liquid medium for 15 days. Low- (1 μg/mL), medium- (10 μg/mL) and high- (100 μg/mL) dose EFs were administrated correspondingly. Alkaline phosphatase (ALP) staining, ALP activity determination, oil red O staining and realtime polymerese chain reaction (RT-PCR) were used to determine the effect of EFs on osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats. Moreover, in order to explore the mechanism of EFs on osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats, Dickkopf-related protein 1 (DKK1) was used in the medium group. Enzymelinked immunosorbent assay (ELISA) and RT-PCR were used to determine mRNA levels of β-catenin, low density lipoprotein receptor-related protein 5 (LRP5) and T cell factor (TCF) protein, known as wnt/β-catenin signal pathway related factors.

RESULTSEFs increased mRNA expression levels of ALP and early osteoblast differentiation factors, such as runt-related transcription factor 2 (Runx2), osteocalcin and collagen I, and decreased mRNA expression levels of fat generation factors, such as peroxisome proliferator activated receptor gamma 2 (PPARγ-2) and CCAAT enhancer-binding protein-α (C/EBPα) in a dose-dependent manner. While osteoblast differentiation factors were down-regulated, fat generation factors were up-regulated when DKK1 was applied. Also EFs up-regulated mRNA expression levels of β-catenin, LRP5 and TCF protein which could be blocked by DKK1.

CONCLUSIONEFs regulate the balance between osteogenic differentiation and adipogenic differentiation in bone marrow stromal cells of ovariectomized rats by activating wnt/β-catenin signal pathway, which may be an important molecular mechanism of EFs on treating postmenopausal osteoporosis.

Adipose Tissue ; cytology ; drug effects ; metabolism ; Animals ; Base Sequence ; Bone and Bones ; cytology ; drug effects ; metabolism ; Cell Differentiation ; drug effects ; DNA Primers ; Enzyme-Linked Immunosorbent Assay ; Epimedium ; chemistry ; Female ; Flavonoids ; pharmacology ; Flow Cytometry ; Mesenchymal Stromal Cells ; cytology ; drug effects ; metabolism ; Rats ; Real-Time Polymerase Chain Reaction ; Signal Transduction ; Wnt Proteins ; metabolism ; beta Catenin ; metabolism