Objective To study in vitro the effects of low-intensity pulsed ultrasound (LIPUS) on the proliferation and differentiation of cultured myoblasts,and to explore the cellular and molecular mechanisms behind any therapeutic effect of LIPUS.Methods Myoblasts were isolated from the skeletal muscles of mice and cultured in vitro.Treatment and control groups of proliferating and differentiating myoblasts were defined.The treatment groups were exposed to LIPUS at 1.5 MHz and a spatial and temporal average intensity of 30 mW/cm2,for 20 min daily,the proliferation group for 6 consecutive days and the differentiation group for 4 consecutive days.The cell proliferation kinetics of the proliferation group were analyzed using flow cytometry.The expression of myogenic regulation factor MyoD and heme oxygenase-1 (HO-1) in the proliferation group,and of myosin heavy chain (MHC) in the differentiation group were examined by immunofluorescent staining.Myoblast fusion indexes were analyzed.Results In the LIPUS treatment groups the proliferating myoblasts had a higher ratio of active cells in the G2 and S phases (19.30％ ±5.14％,37.00％ ±8.72％),compared with the controls (10.33％ ± 1.53％,25.00％ ±4.36％),and the proliferation index increased significantly.The expression of HO-1 was up-regulated,while MyoD staining was unchanged.During the induction of differentiation,the myoblasts of the treatment group fused into smaller myotubes and the myoblast fusion index (18.73％ ± 6.81％) was significantly lower than that of the control group (37.52％ ± 11.23％),while MHC expression did not change markedly.Conclusion LIPUS can promote myoblast proliferation while inhibiting their differentiation,but it does not affect the cells' myogenic properties.HO-1 may be involved in the regulation process.

Objective To study cinical features of patients with idopathic submitral left ventricular aneurysm(ISLVA) and evaluate their global and segmental systolic function as well as diastolic function through echocardiography.Methods Clinical features of eight patients with ISLVA were analysed retrospectively.Standard 2-dimentional and real-time 3-dimentional echocardiography were performed in all the eight cases and other twenty subjects with normal left ventricular(LV) function (defined as control group).Results Two patients were diagnosed as ISLVA due to ventricular arrythmia and the other six case sowing to congestive heart failure.Coronary angiography was normal in all patients.Apical systolic murmur was audible in 6 cases.Four patients received mitral valve surgical repair,one underwent aneurysm resection and radiofrequency ablation,implantable cardioverter defibrillator was implanted in one case,one was only treated by medication.One case died.Patients with ISLVA demonstrated significanly larger left atrium(LA) and LV diameter (both end-diastolic and end systolic),thinner LV posterior wall,and lower LV ejection fraction (LVEF) than controls (P ＜0.05).Indexes of the LV 17 segments time-volume curves including the time to minimal systolic volume(Tmsv) 16-SD,Tmsv 12 SD,Tmsv 6-SD,Tmsv 16-Dif,Tmsv 12-Dif,Tmsv 6-Dif,Tmsv 16-SD％,Tmsv 12-SD％,Tmsv 6-SD％,Tmsv 16-Dif％,Tmsv 12-Dif％,Tmsv 6-Dif％ were significantly higher in patients with ISLVA than those in controls(all P ＜0.05).All patients with ISLVA showed mitral regurgitation and decreased LV diastolic function in varying degrees,five patients accompanied by elevated LA pressure.Conclusions Clinical features of patients with ISLVA are nonspecific.Echocardiography can evaluate systematically their functional and structural abnormalities.

Breast Neoplasms ; genetics ; pathology ; Chromosome Aberrations ; Chromosome Deletion ; Chromosomes, Human, Pair 1 ; Chromosomes, Human, Pair 17 ; Chromosomes, Human, Pair 9 ; Female ; Genome, Human ; Humans ; Nucleic Acid Hybridization ; methods ; Phyllodes Tumor ; genetics ; pathology

Objective To explore the expression and distribution of transient receptor potential cation channel 6(TRPC6)in normal human,mice,rats' renal tissue and the mouse podocyte clone 5(MPC5)for further investigating the relationship between TRPC6 and the protei-nuria-related podocyte molecules.Methods 1.The distributions of TRPC6 in normal human,mice,rats' renal tissue and MPC5 were observed by using the immunochemistry staining.2.The mRNA expression of TRPC6 in mouse renal cortex and differentiated MPC5 was detected by using reverse transcriptase-polymerase chain reaction(RT-PCR).3.The protein expression of TRPC6 in human,mice and differentiated MPC5 was detected by using Western blotting.Results 1.In human kidney,TRPC6 showed a weak staining in glomeruli and a strong staining in renal tubules and vessels.In mice and rats' kidney,TRPC6 showed a strong staining in glomeruli and was mainly distributed along the capillary loops of glomerulus and in mesangium.The positive staining of TRPC6 was observed in MPC5,which was distributed evenly on the cell membrane in differentiated podocytes.2.The specific PCR band of TRPC6 was detected in mouse renal cortex and differentiated MPC5.3.The specific protein band of TRPC6 was detected in normal human,mice renal cortex and differentiated MPC5 with the size of 106.Conclusions The expression of TRPC6 is verified in normal human,mice and rats' kidneys,and in differentiated MPC5.These results will benefit for further exploring the relationship between TRPC6 and the proteinuria-related podocyte molecules.

OBJECTIVETo explore the differences of electroacupuncture (EA) on onset time and symptom improvement for treatment of different types of functional constipation.

METHODSThirty-eight cases of constipation were selected, including 9 cases of constipation-predominant irritable bowel syndrome (IBS-C), 10 cases of slow transit constipation (STC), 10 cases of pelvic floor dyssynergia (PFD) and 9 cases of inadequate defecatory propulsion (IDP). The electroacupuncture was applied at Tianshu (ST 25), Fujie (SP 14), Shenshu (BL 23), Dachangshu (BL 25) and so on in abdominal and lumbosacral area, 5 times per week, 10 times as a treatment course. The onset time, score of clinical symptoms of constipation and improvement of every symptom in each group were compared.

RESULTS(1) The onset time was (1.78 +/- 0.83) days in IBS-C type, (3.11 +/- 1.90) days in IDP type, (4.10 +/- 1.85) days in STC type and (4.30 +/- 2.00) days in PFD type, indicating statistical differences between IBS-C type and STC type, IBS-C type and PFD type (both P < 0.05). (2) Compared before the treatment, the total scores of symptoms on the onset day in each group were all improved (P < 0.05, P < 0.01), and score of IBS-C type was superior to the rest 3 types (P < 0.05, P < 0.01). (3) EA improved desire to defecate or frequency of defecation in each type (P < 0.05, P < 0.01), in which both were improved in STC type and PFD type, and the improvement of defecation frequency was more significant in STC type (P < 0.01). EA relieved unsmooth defecation or pendant-expansion feeling in each type (all P < 0.05), in which both were improved in IBS-C type (both P < 0.05). EA relieved abdominal distension and pain in IBS-C type, STC type and PFD type (all P < 0.05), while its effects were not obvious on defecation difficulty, defecation time and defecation texture (all P > 0.05).

CONCLUSIONThe electroacupuncture for treatment of 4 types of constipation is characterized by rapid onset; the improved symptoms are not identical in the electroacupuncture treatment plan for each type of defecation; the main improvement of symptoms are lied on desire to defecate and frequency of defecation, unsmooth defecation or pendant-expansion feeling and abdominal distension and pain. Meanwhile the improvements of defecation texture, defecation difficulty and defecation time were not signi-ficant.

Adult ; Aged ; Constipation ; etiology ; physiopathology ; therapy ; Defecation ; Electroacupuncture ; Female ; Humans ; Irritable Bowel Syndrome ; complications ; Male ; Middle Aged ; Pelvic Floor Disorders ; complications ; Treatment Outcome ; Young Adult

The Practice Parameters for the Management of Perianal Abscess and Fistula-in-ano from USA(2011) and German guidelines for anal abscess (2012) are based on the evidence and specialists consensus from colorectal field. Standardization of the anal abscess management may simplify the anal fistula treatment. This review is to concepts from other countries and guide the treatment in China.
Abscess ; Anus Diseases ; China ; Drainage ; Humans ; Practice Guidelines as Topic ; Rectal Fistula

This study is to clone the gene of staphylococcal enterotoxins O, obtain recombinant protein (rSEO) and investigate its activity on mice lymphocyte. Staphylococcus aureus O gene is cloned into GST gene fusion vector pGEX-4T-1. The resultant plasmid pGEX-4T-SEO was used to transform E. coLi BL21, where the GST-SEO fusion protein was expressed efficiently. Then SEO was purified by Glutathione Sepharose 4B affinity column and digested with thrombin. The bioactivity of SEO was analyzed by MTT assay on mice lymphocyte and tumor cells. The nucleotide sequence was confirmed to code for the protein correctly, and soluble SEO was expressed efficiently in E. coli BL21 with pGEX-4T-SEO. The protein purified by affinity chromatography resulted to be one single band by SDS-PAGE detection. The MTT assay of the purified rSEO demonstrated that its abilities of stimulating T cells and inhibiting the proliferation of K562, K562-ADM and B16 cells were equivalent to that of SEC in vitro. The expression plasmid pGEX-4T-SEO was constructed and the recombinant superantigen was expressed successfully, which may provide a foundation for the further research of the anticancer activity of SEO.
Animals ; Cell Proliferation ; Cloning, Molecular ; Enterotoxins ; genetics ; immunology ; metabolism ; Escherichia coli ; genetics ; metabolism ; Female ; Genetic Vectors ; Humans ; K562 Cells ; Lymphocytes ; cytology ; Male ; Melanoma, Experimental ; pathology ; Mice ; Mice, Inbred ICR ; Plasmids ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism ; Spleen ; cytology ; Staphylococcus aureus ; genetics ; Superantigens ; genetics ; immunology ; metabolism

OBJECTIVETo assess the therapeutic effects and acting specialty of acupuncture and moxibustion for treatment of slow-transmission constipation (STC).

METHODSA clinical pathway was created and Patient-Reported Outcomes (PROs) method was taken for efficacy evaluation dominantly. Acupuncture and moxibustion were applied to 30 STC patients on two groups of acupoints in alternation, with acupoints of Tianshu (ST25), Daheng (SP15), Fujie (SP14), Qihai (RN6), Guanyuan (RN4), Zusanli (ST36), and Shangjuxu (ST37) as group 1; Dachangshu (BL25), Shenshu (BL23), Baliao (BL31, BL32, BL33, BL34), and Sishencong (Ex-HN1) as group 2. Deep needling was applied on acupoints of abdominal and back region and moxibustion was put on Sishencong, 20 times as one course. The therapeutic effect was assessed based on the Patient Assessment of Constipation Quality of Life Questionnaire (PAC-QOL) method by comparing the Bristol scoring on quality of stool, awareness and frequency of defecation, degree of abdominal fullness sensation, and patients' quality of life (QOL) at different time points, i.e., pre-treatment, after 1, 2 and 3 weeks of treatment.

RESULTSCompared with the condition of pre-treatment, after 1, 2 and 3 weeks of treatment, cases using irritant laxative or glycerin enema reduced, with quality of stool normalized, scores for awareness and frequency of defecation increased, and abdominal fullness sensation lessened (all P < 0.01); both the total score and scores on the 4 domains of QOL (malaise, psychosocial complaint, anxiety and interest related to constipation, and satisfaction) were obviously reduced. No adverse event occurred during the treatment course. Therapeutic effectiveness assessment at the end of treatment showed that 13 patients were cured, 9 improved and 8 treated in vain; the corresponding cases assessed at 1 month after treatment were 10, 7, 13; and those at 3 months after were 7, 8, 15, respectively.

CONCLUSIONEstablishing a clinical pathway is helpful to patients in their continuous diagnosis and treatment. Using PROs method to evaluate the therapeutic effect could clearly reveal the superiority of acupuncture and moxibustion in improving symptoms and QOL in patients of STC.

Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Colon ; physiopathology ; Constipation ; physiopathology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Moxibustion ; Treatment Outcome ; Young Adult

OBJECTIVETo prepare and identify monoclonal antibodies against staphylococcal enterotoxin I (SEI).

METHODSSpleen cells obtained from mice immunized with the SEI protein were fused with the myeloma cells (SP2/0). Hybridomas were screened by enzyme-linked immunosorbent assay (ELISA) and the stable monoclonal hybridomas were isolated by limiting dilution at least three times. The characters of purified monoclonal antibodies were identified by indirect ELISA and Western blotting.

RESULTThe monoclonal antibodies secreted by two hybridomas 8F7 and D8 belonged to IgG(2b) and IgG(1) subtypes. Both had high titer and specificity with no cross reaction to SEG, SEE and SEC.

CONCLUSIONThe monoclonal antibodies against SEI has been successfully prepared and identified in this study.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Enterotoxins ; immunology ; Hybridomas ; secretion ; Immunoglobulin G ; biosynthesis ; immunology ; Mice ; Mice, Inbred BALB C ; Spleen ; cytology ; Staphylococcus aureus ; immunology

The filtrate of Staphylococcus aureus culture has been used in an ampule form named as staphylococcal enterotoxin C injection for cancer therapy in clinic for ten years in China and proved to be effective. The active constituent of three kinds of injections is claimed to be staphylococcal enterotoxin C2 (SEC2), and the content of SEC2 is used as quality control. However, the correct content of SEC2 was not known and the relative amount of SEC2 was very low because of the complicated components of the filtrate. In this research, we established a proper ELISA system for the detection of SEC2 in staphylococcal enterotoxin C injection, which will improve the quality control of the injection. We produced and identified polyclonal and monoclonal antibodies of SEC2 and established BA-ELISA method based on the method of sandwich ELISA. It was found that the BA-ELISA method had good specificity, sensitivity and reproducibility, and being able to detect SEC2 at concentration from 2 to 20 ng x mL(-1), with an average CV value of 5.08%. The SEC2 content in staphylococcal enterotoxin C injection was calculated. There is some difference between the actual and labeled contents in the injections.
Animals ; Antibodies, Bacterial ; biosynthesis ; immunology ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antineoplastic Agents ; administration & dosage ; analysis ; immunology ; Enterotoxins ; administration & dosage ; analysis ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Hybridomas ; secretion ; Injections ; Mice ; Mice, Inbred BALB C ; Quality Control ; Rabbits ; Staphylococcus aureus ; chemistry