Acupuncture Therapy ; Adolescent ; Humans ; Male ; Superior Mesenteric Artery Syndrome ; therapy

OBJECTIVETo determine the regulatory effects of the circadian clock gene Per1 on cell cycle-related genes and its influence on the proliferation, apoptosis, cycle, and tumorigenicity in vivo of human oral squamous cell carcinoma SCC15 cells.

METHODSThree groups of the short hairpin RNA (shRNA) of lentivirus recombinant plasmids were designed against the RNA of Per1 and then transfected to the SCC15 cells. The optimum interference group was screened through Western blot and quantitative real-time PCR (qRT-PCR) and assigned as the experimental group. The transfected lentivirus plasmid without an interference effect on any gene was set as the control group (Control-shRNA). Untreated SCC15 cells were set as the blank group. The mRNA expressions of cell cycle-related genes, namely, Per1, p53, Cyclin D1, Cyclin E, Cyclin A2, Cyclin B1, CDK1, CDK2, CDK4, CDK6, p16, p21, Wee1, cdc25, E2F, and Rbl1 in each group were detected through qRT-PCR. The cell proliferation, apoptosis, and cell cycle distribution in each group were evaluated through flow cytometry. The cells of the experimental group and the blank group were subcutaneously inoculated in nude mice to observe tumorigenesis.

RESULTSThree groups of Per1-shRNA lentivirus plasmids were constructed successfully. Among the groups, the Per1-shRNA- I group exhibited the highest interference effect, as indicated by qRT-PCR and Western blot analysis. As such, this group was set as the experimental group. The mRNA expression levels of CyclinD1, CyclinE, CyclinB1, CDK1, and Wee1 gene in the Per1-shRNA-I group were significantly higher than those in the Control-shRNA group and the SCC15 group (P < 0.05). By contrast, the mRNA expression levels of p53, Cyclin A2, p16, p21, and cdc25 in the Per1-shRNA-I group were significantly lower than those in the Control-shRNA group and the SCC15 group (P < 0.05). The mRNA expression levels of each gene between the Control-sLRNA group and the SCC15 group did not significantly differ (P > 0.05). The mRNA expression levels of CDK2, CDK4, CDK6, E2F, and Rb1 did not significantly differed in the three groups (P > 0.05). The proliferation index of the Perl-shRNA-I group was significantly higher than those of the Control-shRNA group and the SCC15 group (P < 0.05). The apoptosis index of the Per1-shRNA-I group was significantly lower than those of the Control-shRNA group and the SCC15 group (P < 0.05). The number of S-phase cells in the Per1-shRNA-I group was significantly lower than those of S-phase cells in the Control-shRNA group and the SCC15 group (P < 0.05). The number of G2/M-phase cells in the Per1-shRNA-I group was significantly higher than those of G2/M-phase cells in the Control-shRNA group and the SCC15 group (P < 0.05). Conversely, the proliferation index, apoptotic index, and cell cycle distribution of the cells in the Control-shRNA group did not significantly differ from those of the SCC15 group (P > 0.05). The tumorigenic ability in vivo was significantly enhanced in the Per1-shRNA-I group (P < 0.05).

CONCLUSIONPer1 is an important tumor suppressor gene. Per1 can regulate a large number of downstream cell cycle-related genes. The alteration of its expression can affect cell cycle progression, proliferation, apoptosis imbalance, and tumorigenic ability in vivo. Further studies on Per1 may elucidate cancer development and provide novel effective molecular targets for cancer treatment.

Animals ; Apoptosis ; Carcinoma, Squamous Cell ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Circadian Clocks ; genetics ; Cyclin D1 ; Humans ; Mice ; Mice, Nude ; Mouth Neoplasms ; Period Circadian Proteins ; genetics ; Plasmids ; RNA, Small Interfering ; Real-Time Polymerase Chain Reaction ; Transfection

Objective To explore the effect of electroacupuncture at Hegu (LI 4) on the expressions of P2X2 and P2X3 receptors in experimental pulpalgia rats.Method Forty-two male SD rats were randomized into a normal group (group N), a control group (group C), a pulpalgia model group (group M), an antagonist group (group A), an electroacupuncture group (group E), and an antagonist+electroacupuncture group (group AE), 7 rats in each group. Group N didn’t receive any interventions; group C received injection of normal saline into pulp cavity of the same dose as the injection in group M, and the cavity was then blocked by dental fillings 5-6 min later; in group M, maxillary first and second molar teeth were drilled (drill bit of 1 mm in diameter) to expose pulp and lipopolysaccharide (LPS) solution 5μg/μL was injected into the holes (1~3μL for each hole), and the holes were then covered by dental fillings 5-6 min later; group A received the same modeling method as that in group M, but A-317491 was injected together with LPS (0.5 mg/kg); group E received electroacupuncture at bilateral Hegu (LI 4) with needles retained for 30 min, once a day, totally for 3 times; group AE received the same electroacupuncture intervention after receiving the same treatments as that in group A. The rats’ behaviors and weight were observed for 30 min after intervention each day. The rats were sacrificed on the 4th day, and the mRNA expressions of P2X2 and P2X3 receptors andβ-actin in trigeminal ganglion were detected by using RT-PCR. The mRNA expressions were then compared among the groups.Result The behavioral changes in group M, E, and AE were more significant than that in group C and N (P<0.01); the behavioral changes in group A, E, and AE were less significant than that in group M (P<0.01). The weight in group C was significantly lower than that in group N (P<0.01); the weights in group M, A, E and AE were significantly lower than that in group C and N (P<0.01); the weights in group E and AE were significantly higher than that in group M and A (P<0.01,P<0.05); the weight in group A was slightly higher than that in group M (P<0.05); the weights in group AE was significantly higher than that in group E (P<0.01). The mRNA expressions of P2X2 receptor in group M, A, and AE were significantly higher than that in group N and C (P<0.01); the mRNA expressions of P2X2 receptor in group A, E, and AE were lower than that in group M (P<0.05); the mRNA expression of P2X2 receptor in group A was lower than that in group E (P<0.05); the mRNA expression of P2X2 receptor in group E was higher than that in group AE (P<0.05). The mRNA expressions of P2X3 receptor in group M, A, and E were significantly higher than that in group C (P<0.05) and group N (P<0.01); the mRNA expressions of P2X3 receptor in group A, E, and AE were significantly lower than that in group M (P<0.01); the mRNA expression of P2X3 receptor in group AE was markedly lower than that in group E (P<0.01).Conclusion The expressions of P2X2 and P2X3 receptors in trigeminal ganglion were increased in LPS-induced pulpalgia rats. Electroacupuncture at Hegu (LI 4) and injection of A-317491 both can down-regulate the mRNA expressions of P2X2 and P2X3 receptors, which is plausibly the action mechanism of electroacupuncture at Hegu (LI 4) in analgesia.

Objective To investigate the effect of penehyclidine hydrochloride (PHCD) on tramadol dependence and c-fos,△ FosB and M5 receptor expression in relevant brain regions in rats.Methods Thirty male adult SD rats weighing 180-220 g were randomly assigned to 3 groups (n =10 each):control group (group C),tramadol dependence group (group T) and PHCD group (group P).Tramadol dependence was induced by subcutaneous 10 mg/kg once a day for 7 consecutive days in groups T and P.PHCD 1.5 mg/kg was injected intraperitoneally on day 8 in group P,while in groups C and T the equal volume of normal saline was injected intraperitoneally instead of PHCD.The rats underwent conditioned place perference test at 30 min after intraperitoneal injection.The time spent in drug-paired side (gray area) was recorded.The rats were sacrificed after the conditioned place perference testand the brain was removed.The relevant brain regions (ventral tegmental area,prefrontal cortex,nucleus accumbens )were separated for determination of c-fos,△ FosB expression by Western blot and M5 receptor mRNA expression by RT-PCR.Results Compared with group C,the time spent in the drug-paired side (gray area) was significantly prolonged,and c-fos,△FosB and M5 receptor mRNA expressions were up-regulated in group T,△FosB and Ms receptor mRNA expressions were down-regulated in group P ( P ＜ 0.05 or 0.01 ).There was no significant difference in time spent in the drug-paired side (gray area) and c-fos expression between groups C and P( P ＞ 0.05).Compared with group T,the time spent in the drug-paired side (gray area) was significantly shortened,and c-fos,△ FosB and M5 receptor mRNA expressions were down-regulated in group P (P ＜0.01).Conclusion PHCD can significantly inhibit tramadol dependence by down-regulating c-fos,△FosB and M5 receptor expression in relevant brain regions.

Objective to investigate the efficacy of moxibustion treatment to rheumatoid arthritis(RA) patients and the impact to blood levels of rheumatoid factor(RF) ,C reactive protein(CRP) ,and interleukin-6(IL-6) .Methods RA patients were divided in-to treatment group and comfort group .The two groups were treated by moxibustion and sham-moxibustion for 2 courses ,respec-tievely .Results Overall clinical efficacy rate was higher in treatment group than in comfort group(P<0 .01) .The arthralgia ,ar-throcele ,tenderness and its degrees ,and morning stiffness time were significant improved in both groups(P<0 .05) ,and the indexes were better in treatment group than comfort group(P<0 .05) .The blood levels of RF ,IL-6 and CRP were reduced after treatment in both groups(P<0 .05) ,and which of treatment group reduced more than those of comfort group(P<0 .05) .Conclusion Moxi-bustion treatment can significantly improve clinical outcomes of RA and markedly reduce the blood levels of RF ,IL-6 and CRP of RA patients .

Objective To summary the experience of AngioJet mechanical thrombectomy for the treatment of acute deep venous thrombosis (DVT) of lower extremity.Methods Clinical data of 28 patients suffering from acute DVT of lower extremities treated by AngioJet mechanical thrombectomy were analyzed retrospectively from October 2013 to February 2015.Venous recanalization was graded by a thrombus score based on pre-and post-treatment venography.Follow-up was performed by Doppler ultrasound and clinical evaluation.Results Twelve patients were planted temporary vena cava filters.The average length of hospitalization was (3.9 ± 1.1) days.The average operation time was (1.2 ± 0.4) hours.The dosage of urokinase was (320 ± 120) × 104 U and heparin was (46 ± 10)mg during surgery.The average reduction of hemoglobin was (7.5 ± 2.6) g/L.Two patients (Grade Ⅰ thrombolysis) were converted to catheter-directed thrombolysis (CDT).There were 10 (35.7％) cases achieving grade Ⅱ and 16 (57.1％) cases achieving grade Ⅲ,respectively.Technique success rate were 92.9％.Minor bleeding events occurred in 2 patients.Nine (32.1％) patients had stents implanted.One year follow-up found patent deep venous in 26 (92.9％) patients and stent patency rate was 88.9％ (1/9).Two (7.1％) cases developed mild postthrombotic syndrome (PTS).Conclusion AngioJet is a safe and effective tool to treat acute DVT accelerating thrombolytic therapy,and reducing bleeding complications.

Ammonium perchlorate (AP), mainly used as solid propellants, was reported to interfere with homeostasis via competitive inhibition of iodide uptake. However, detailed mechanisms remain to be elucidated. In this study, AP was administered at 0, 130, 260 and 520 mg/kg every day to 24 male SD rats for 13 weeks. The concentrations of iodine in urine, serum thyroid hormones levels, total iodine, relative iodine and total protein, and malondialdehyde (MDA), superoxide dismutase (SOD) and catalase (CAT) activity in thyroid tissues were measured, respectively. Our results showed that high-dose perchlorate induced a significant increase in urinary iodine and serum thyroid stimulating hormone (TSH), with a decrease of total iodine and relative iodine content. Meanwhile, free thyroxine (FT4) was decreased and CAT activity was remarkably increased. Particularly, the CAT activity was increased in a dose-dependent manner. These results suggested that CAT might be enhanced to promote the synthesis of iodine, resulting in elevated urinary iodine level. Furthermore, these findings suggested that iodine in the urine and CAT activity in the thyroid might be used as biomarkers for exposure to AP, associated with thyroid hormone indicators such as TSH, FT4.

ObjectiveTo explore the normal range of the volume of internal capsules in Chinese adults of the Han nationality and its relationship with age,body habitus,and craniocerebral volume.Methods One thousand healthy volunteers (age range =18 to 80 years) were divided into 5 groups according to their age;Group A ( 18 to 30 years old),group B (31 to 40 years),group C (41 to 50 years),group D (51 to 60 years),and group E (61 to 80 years).Each group consisted of 100 males and 100 females.MR imaging was performed in all of the volunteers using T1 weighted three-dimensional nagnetization prepared rapid acquisition gradient echo sequence. After three dimension data reconstruction,the volumes of bilateral internal capsules were manually measured. The volumes of bilateral internal capsules were compared by paired sample t test.The internal capsule volumes were compared between male and female by independent sample t test,and the differences among 5 age groups were compared by one-way ANOVA.The relationship between the volumes of internal capsule and age,body habitus or cerebral volume were analyzed using bivariate correlation.ResultsThe left and right internal capsule volumes were (2809 ± 393) and (2677 ± 343 ) mm3 respectively.The left internal capsule volumes were significantly larger than that of right (t =12.078,P ＜ 0.05 ).The left and right side of internal capsule volumes in male were (2863 ± 396) and (2744 ±358) mm3 respectively,and (2754 ±385) and (2609 ±314) mm3 in female.The left and right internal capsule volumes were larger in males than in female (t =1.982,2.851 ;P ＜ 0.05 ).The left internal capsule volume of the 5 age groups were ( 3273 ± 361 ),( 2943 ± 299 ),( 2777 ± 255 ),( 2607 ± 199 ),(2444 ±213) mm3,and the right were (2993 ± 361 ),(2814 ± 270),(2682 ± 239),(2543 ± 219),(2351 ±210) mm3.There were significant differences among 5 age groups between left and right internal capsule volume ( F =55.244,34.493 ; P ＜ 0.05 ).There was significant negative correlation between the volume of left and right internal capsule and age ( r =- 0.718,- 0.637 ; P ＜ 0.05 ).Conclusions1.5 T MR scanner can be used to accurately measure the internal capsule volumes.There is a significant negative correlation between age and internal capsule volumes.

Objective To explore the risk factors of inguinal metastasis in squamous cell carcinoma of the penis, screening lymph node metastasis high-risk patients. Methods The clinical and pathological data of 81 consecutive patients with squamous cell carcinoma of the penis were analyzed retrospectively. Age at presentation ranged from 27 to 81 years with a median of 49 years. Course of disease within one year of patients with 46 cases (56.8%), 1 year above 35 eases (43.2 %). Seventyfive patients underwent bilateral inguinal lymph node dissection, and 6 patients had unilateral inguinal lymph node dissection. Clinical stage of the primary tumor was assigned according to the 2002 TNM staging system. Variables included patients' age, redundant prepuce and/or phimosis, tumor site,size, number, macroscopic growth pattern, histological grade, inguinal physical examination and the size of inguinal lymph nodes. Results Of the 81 patients, 42 (51.9%) were staged as pN+ and 39 (48. 1%) as pN0. Metastases occurred in 32.0% (16/50) of G1, 78.3% (18/23) of G2 and 100. 0%(8/8) of G3 cases, with significant differences among them (P= 0. 015). According to the inguinal lymph node physical examination results, 63 were staged as clinically node-positive (cN+) and 18 as clinically node-negative (cN0). Metastases occurred in 63. 5% (40/63) of cases of cN+, as compared with 11.1% (2/18) of cases of cN0(P=0. 012). At a median follow up of 40 months (ranged 2-127 months), the 5-year disease free survival rates for positive and negative inguinal lymph nodes metastasis were 71.4% and 92.3%, respectively (P=0. 005) , and the 5-year cancer specific survival rates were 79.0% and 91.4%, respectively (P=0.001). Conclusions Inguinal physical examination and histological grade were the strongest predictors of inguinal metastasis. The patients with inguinal lymph nodes metastasis have lower 5-year disease free survival rates and cancer specific survival rates,and should receive positive treatment measures.

Objective:To evaluate the clinical research papers published in recent years about tuberculosis , to understand the ethical review in biomedical journals in our country .Method:We analyzed clinical papers published in two journals that are famous in the field of tuberculosis since 2010 .Results:There were 33 articles included in our study .The number of informed consent is higher than that of ethical review .The descriptions of informed con-sents were not standardized in 12%articles.9%of articles did not mention the name of the ethics committee .Con-clusion:The attention degree to ethics of some editors and researchers should be improved .Pay more attention to medical ethical education , improve cognitive level of ethical issues , ensure the interests of the participants and im-prove the quality of research .