Objective To establish a rapid assay for genotyping of IL-6-597G/A and-572G/C polymorphisms by duplex real-time PCR assay.Methods One pair of primers and two pairs of fluorescent hybridization probes had been used to genotype two mutation sites in the promoter region of IL-6 gene by fluorescent resonance energy transfer and melting curves.Results Duplex real-time PCR method for genotyping of two mutation sites simultaneously had been developed and 123 health people samples were analyzed by this new method.The results showed that three genotypes were found in IL-6 gene-572G/C polymorphisms.They were GG,GC and CC genotypes.IL-6 gene promoter-597G/A polymorphism analysis showed 4 cases displayed GA genotype and other possessed GG genotype.No AA genotype had been found.Conclusions Duplex real-time PCR method is simple and fast.It provided an accurate and economic method which is suitable for clinical gene genotyping.

Professor JIA Chunsheng believes that "Dongyuan's acupuncture therapy" proposed by GAO Wu could not entirely reflect LI Dongyuan's academic thought of acupuncture and moxibustion. Hence, we collected all of the academic works of LI Dongyuan and extracted the texts relevant with acupuncture-moxibustion therapy, titled as Dongyuan's acupuncture-moxibustion therapy. This process contributed to the deeper study on Dongyuan's acupuncture and moxibustion academic thought. By the influence of Dongyuan's acupuncture-moxibustion academic thought, in association with his clinical experience for dozens of years, Professor JIA Chunsheng proposed that acupuncture and moxibustion should pay attention to the regulation and tonification of spleen and kidney. In clinical practice, Zhongwan (CV 12), bilateral Tianshu (ST 25), Qihai (CV 6) and Guanyuan (CV 4) are commonly selected to consolidate the root, cultivate the primary and treat the spleen and kidney simultaneously. The locations of the above selected points are distributed like a star. This "star-like point selection method" points to the importance of the spleen and kidney (the primary qi). For many diseases, especially the disorders of internal injury, the star-like acupuncture therapy is commonly used and the relevant acupoints are supplemented. The good clinical efficacy has been achieved.
Acupuncture Points ; Acupuncture Therapy ; history ; methods ; China ; History, Ancient ; Humans ; Moxibustion ; history ; methods

Objective:To investigate the expression of wild type estrogen receptor(wER)and the ex-on-5 deleted ER(variant ER,vER)in human hepatocellular carcinoma(HCC)samples,and thereafteranalyze the possibility of HCC treatment by endocrine therapy.Methods:The mRNA expressions of wERand vER were analysed from 28 cases of HCC by RT-PCR.The expression of ER at the protein level wasdetected by immunohistochemistry(IHC).Results:IHC results showed that 39.3% of the HCC speci-mens expressed ER.The mRNA of wER was detected in 89.3%(25/28)of the HCC specimens whilethat of vER was detected in 96.4%(27/28).Twenty four out of 28 HCC cases(85.7%)expressedboth wER and vER.One out of 28 patients(3.5%)expressed only wER whereas 3 patients out of 28(10.7%)expressed vER only.Conclusion:Ninety six percent(27/28)of the HCC patients expressedvER,which suggests that the expression of vER is an important event in the development of HCC.

Objective:To analyze the mean levels of skeletal muscle mass and strength in elderly patients with type 2 diabetes mellitus(T2DM), and to investigate the effects of chronic inflammatory factors and oxidative stress on them.Methods:A cross-sectional study was conducted on 120 patients with T2DM aged over 60 years and 126 elderly patients without diabetes(the control group). Skeletal muscle mass, strength and serum levels of chronic inflammatory factors interleukin-6(IL-6), tumor necrosis factor-α(TNF-α)and 8-hydroxy-2′-deoxyguanosine(8-OHdG)were determined, and their effects on skeletal muscle mass and strength in elderly patients with T2DM were analyzed.Results:Compared with the control group, grip strength decreased in elderly patients with T2DM(25.03±7.85)kg vs.(29.52±7.73)kg( P<0.01), and skeletal muscle mass decreased(21.36±5.46)kg vs.(22.01±5.22)kg with no significant difference( P>0.05). Serum levels of 8-OHdG were higher in elderly patients with T2DM than in the control group(3.08±0.26)ng/L vs.(2.59±0.16)ng/L( P<0.01). Correlation and regression analysis results showed that 8-OHdG was an influencing factor for muscle strength in elderly patients with T2DM( R2=0.457)and that height and weight could be influencing factors for skeletal muscle mass in elderly patients with T2DM( R2=0.822). Conclusions:Skeletal muscle mass and strength decline in elderly T2DM patients, probably as a result of increased levels of oxidative stress.These findings may serve as evidence for sarcopenia intervention in elderly T2DM patients.

OBJECTIVETo evaluate the erectile and ejaculatory function of sacral tumor patients after sacral nerve root resection and investigate the relationship of erectile and ejaculatory dysfunction (EED) with the level of sacral nerve injury.

METHODSThis retrospective study included 47 male patients aged 16 to 63 (32.6 +/- 6.8) years treated by sacral tumor resection between January 2008 and August 2013. According to the levels of the sacral nerve roots spared in surgery, the patients were divided into four groups: bilateral S1-S3 (n=16), unilateral S1-S3 (n=21), unilateral S1-S2 (n=6), and unilateral S1 (n=4). The patients were followed up for 12 to 41 (27.2 +/- 10.9) months by questionnaire investigation, clinic review, and telephone calls about their erectile and ejaculatory function at 3, 6 and 12 months after surgery and in August 2013.

RESULTSIn the bilateral S1-S3 group, the incidence rates of EED were 31.25% (5/16), 25% (4/16), and 12.5% (2/16) at 3, 6, and 12 months respectively after surgery, with recovery of erectile and ejaculatory function in August 2013. The incidence rates of EED in the unilateral S1-S3 group were 85.71% (18/21), 71.43% (15/21), 52.38% (11/21), and 42.86% (9/21) at 3, 6 and 12 months and in August 2013, respectively; those in the unilateral S1-S2 group were 100% (6/6), 83.33% (5/6), 83.33% (5/6), and 66.67% (4/6) at the four time points; and those in the unilateral S1 group were all 100% (4/4). No statistically significant differences were found in the incidence rate of EED among the patients of different ages or tumor types (P > 0.05).

CONCLUSIONThe incidence of postoperative EED in male patients treated by sacral tumor resection is closely related to the mode of operation. Sparing the S3 nerve root at least unilaterally in sacral tumor resection is essential for protecting the erectile and ejaculatory function of the patient.

Adolescent ; Adult ; Ejaculation ; physiology ; Erectile Dysfunction ; epidemiology ; etiology ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Organ Sparing Treatments ; Peripheral Nervous System Neoplasms ; surgery ; Postoperative Complications ; epidemiology ; Postoperative Period ; Retrospective Studies ; Sacrum ; Spinal Nerve Roots ; injuries ; surgery ; Surveys and Questionnaires ; Young Adult

Abstract: Schistosomiasis is a serious major parasitic disease that threatens human life and health. A better understanding of the mechanism of host-schistosome interactions is the key to designing new prevention and control strategies. MicroRNAs (miRNAs) are endogenous small non-coding RNA molecules, which lead to the degradation of the target messenger RNA (mRNA) or inhibition of its translation in a sequence-specific manner. Both schistosome and its host produce miRNAs, which can be secreted by extracellular vesicles (EVs). There is accumulating evidence that miRNAs from schistosome can be taken up by host cells, and finely manipulate the phenotype of host cells for their survival or pathogenesis in a cross-species manner, even inhibiting the growth and metastases of hepatoma cells. It is still unknown whether host free miRNAs can be taken up by schistosome, but this phenomenon is highly probable. miRNA-mediated cross-species regulation has emerged as a novel mechanism for host-schistosome interactions, and this review summarizes the advances in this regard.

This study was aimed to explore the correlation between IL-6 gene promoter polymorphisms and coronary heart disease (CHD) by investigating the polymorphisms (-572G/C, -597G/A) in IL-6 gene promoter area, body mass index (BMI), inflammatory factors and other biochemical parameters in Han nationality of North China. The genotypes of IL-6 gene promoter-572G/C, -597G/A were detected by fluorescent probe hybridization with fluorescent resonance energy transfer and melting curve techniques in 194 CHD patients and 123 healthy people as control. The effects of genotype on plasma lipids, apoproteins, high sensitive C-reactive protein (hsCRP) and BMI were also studied. Logistic regression was performed to observe the risk factors of CHD. The results indicated that genotype of IL-6 gene promoter -597G/A polymorphism in 7 cases were GA and were GG in others, whereas no AA genotype had been found and no associations between polymorphism of IL-6 gene -597G/A, BMI and inflammatory factors were found. No differences had been found between the frequencies of IL-6 gene -572G/C genotypes and alleles in CHD and control group. However, significant difference was found between the G allele carrier (GG+GC) and non-G allele carrier (CC) of CHD and control group (p=0.0425). In the control group, median levels of systolic blood pressure of G allele carrier were significant higher than non-G allele carrier (p=0.02). Among all the subjects, median levels of BMI, hsCRP and systolic blood pressure in the group of G allele carrier were significantly higher than that in the group of non-G allele carrier, p values were 0.026, 0.022, 0.005 respectively. Multivariate logistic regression analysis showed that age, triglyceride, sex, high blood pressure, apoprotein C2, cholesterol and lipoprotein-a) were the risk factors for CHD, and apoprotein A1 was a protective factor. The G allele of IL-6 gene -572G/C has been not found to be a risk factor for CHD. It is concluded IL-6 gene -597G/A polymorphism is not correlated with the susceptibility to CHD; IL-6 gene -572G/C polymorphism may be correlated with the susceptibility to CHD in Han nationality of North China, the mechanisms may be related with the changes of BMI, hsCRP and blood pressure levels resulted from the polymorphism of IL-6 -572G/C.
Adult ; Aged ; Body Mass Index ; C-Reactive Protein ; metabolism ; Coronary Disease ; etiology ; genetics ; Disease Susceptibility ; Female ; Genotype ; Humans ; Interleukin-6 ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; Promoter Regions, Genetic ; genetics ; Risk Factors

OBJECTIVETo explore the effect of rosiglitazone on the activity of signal transducer and activator of transcription 1 in rats with severe acute pancreatitis.

METHODSFifty-four male Wistar rats were randomly allocated into three groups (n = 18). SO group: sham-operated animals served as control, operation was executed and sodium chloride but not sodium taurocholate was injected. SAP group: SAP was induced by the retrograde injection of 5% sodium taurocholate into the biliopancreatic duct. ROSI group: same as SAP group, but rosiglitazone (6 mg/kg) was administered intravenously 30 min before operation. Rats in each group were sacrificed at 3,6 and 12 h after operation. The levels of serum amylase and histologic scores of pancreatic tissue were measured. The expression of TNF-alpha mRNA in pancreatic tissue were determined by reverse transcription-polymerase chain reaction (RT-PCR). The expression of phosphorylated STAT1 in pancreatic tissue was assayed by immunohistochemistry.

RESULTSCompared to SO group, the levels of serum amylase and phosphorylated STAT1, TNF-alpha mRNA and histologic scores of pancreatic tissue were significantly elevated at the same time points after SAP (P < 0.01). The levels of these detection in ROSI group were lower than those of the SAP group at the same time points (P < 0.05), but higher than SO group (P < 0.05).

CONCLUSIONSSTAT1 was activated in severe acute pancreatitis. Rosiglitazone has a protective effects in rats with severe acute pancreatitis. The mechanism of its protective effects maybe that it inhibits the activation of JAK/STAT pathway, which can down-regulate the expression of TNF-alpha mRNA and block the the inflammatory cascade partially.

Acute Disease ; Animals ; Disease Models, Animal ; Male ; Pancreas ; metabolism ; pathology ; Pancreatitis ; drug therapy ; metabolism ; pathology ; Random Allocation ; Rats ; Rats, Wistar ; STAT1 Transcription Factor ; metabolism ; Thiazolidinediones ; pharmacology

AIMTo study the effects of stress on the cognitive function and physical fitness and its biological mechanisms, provide scientific basis for seeking protective measures to reduce stress-induced damage.

METHODSThe model of restraint stress was adopted in our experiment. Step-down test and exhaustive swimming test were used to measure the learning and memory function and physical fitness in mice, respectively. The contents of GC in plasma, hippocampus LTP, ECG, myocardial ultra-structure, cell apoptosis rate and the level of Hsp70 expression of myocardium in rats were detected.

RESULTSCompared with control group, the impairment of learning and memory function and decline of exercise tolerance were observed in restraint stress group. The elevation in plasma GC levels, ECG abnonrmality, and cell apoptosis rate were also observed under restraint stress. Furthermore, myocardial structure was damaged, and myocardial Hsp70 expression and hippocampus LTP were suppressed in restraint stress group than those in the control.

CONCLUSIONStress may cause the neuro-endocrine dysfunction and homeostasis disorder, and then, lead to the cognitive function and physical fitness damage consequently.

Animals ; Cognition ; physiology ; HSP70 Heat-Shock Proteins ; metabolism ; Hippocampus ; physiology ; Long-Term Potentiation ; physiology ; Male ; Mice ; Myocardium ; metabolism ; Physical Fitness ; physiology ; Random Allocation ; Rats ; Rats, Wistar ; Restraint, Physical ; Stress, Physiological ; physiology

AIMTo construct the RNAi eukaryotic vector of inhibitory member of the prohibitin (PHB-1) gene and observe the interfering effect in HEK293 cell line after the vector transfection.

METHODSThe specific Mi RNA sequence was designed according to the PHB-1 sequence in GenBank, complementary single-strand DNA oligonucleotides were designed and synthesized, and annealed the single-stranded oligonucleotides to generate a double strands oligonucleotides , cloned the oligonucleotides into pcDNATM6.2-GW/EmGFP-MiR-PHB to obtain an entry clone and then sequence analysis was performed. The recombinant plasmid was transfected into HEK293 cell by liposome. PHB-1 expression was detected by Western blotting.

RESULTSThe DNA sequence of interest clone to the vector was constructed to generate an entry clone and an expression clone successfully, which were proved by sequence determination. Western blotting analysis demonstrated that PHB-1 MiR RNA expression construction could suppress the expression of PHB-1.

CONCLUSIONA RNAi eukaryotic vector containing prohibitin gene was successfully constructed.

Genetic Vectors ; genetics ; HEK293 Cells ; Humans ; MicroRNAs ; biosynthesis ; genetics ; RNA Interference ; RNA, Small Interfering ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Repressor Proteins ; biosynthesis ; genetics ; Transfection