Six 89bp primers were designed on the base of the cDNA sequence encoding the human leptin reported on the NCBI. The synthetic gene with 464bp encoding rhLep was obtained by SOE ( splicing by overlap extension) PCR. The expression vector pET22b(+)/rhLep was constructed and transformed into E. coli BL21 (DE3). The rhLep protein was expressed as inclusion bodies with the yield of more than 50% of total bacterial proteins after IPTG induction. The rhLep protein, which has a molecular weight about 16kD, was purified by Ni2+ affinity chromatography column and identified by SDS-PAGE. The MTT Assay shows that rhLep promotes EC304 cells growth at the low concentration of 10ng/mL to 30 ng/mL, and rhLep appears cytotoxic to EC304 cells with the high dose of 50ng/mL to 225ng/mL. The viability of EC304 cells decreases to 1.2% with the concentration of 225ng/mL of rhLep. The massive apoptosis of rhLep on EC304 cells is observed by AO-staining under fluorescent microscope. All these results would lay the foundation for the further study of its biological functions in vitro and in vivo.
Apoptosis ; drug effects ; Escherichia coli ; genetics ; Humans ; Leptin ; genetics ; pharmacology ; Recombinant Proteins ; biosynthesis ; isolation & purification ; pharmacology

Ten acupuncture-related articles were published in The Journal of the American Medical Association (JAMA) between 1998 and 2017. Five studies showed positive results in terms of the effectiveness of acupuncture/Chinese medicine (CM); five studies showed negative results. This article summarizes the acupuncturerelated clinical trials published over the last 20 years in JAMA, and addresses what seems to be a fundamental ambivalence in Western medical journals regarding the scientific validity of acupuncture/CM. As yet there has been no consensus on the role of acupuncture in healthcare in Western countries. This is hardly surprising, considering the conflicting evidence found in published studies. Skepticism regarding acupuncture/CM is largely grounded in the fact that an accurate model for assessing the true clinical effects of acupuncture has yet to be created. This article discusses some of the pitfalls which result from applying Western-based scientific principles to CM, and suggests that in many cases, "negative" studies have been misinterpreted. The clinical experience of acupuncture practitioners is often in direct conflict with many of the negative conclusions published in journals. We are in need of an accurate model for sham and placebo treatments, and must analyze all published studies for design flaws and faulty conclusions.

The opioid epidemic has become a signifificant public health crisis in the United States of America. This crisis has elicited a response at high levels of governmental and health care organizations including the American College of Physicians, the Food and Drug Administration, the Joint Commission, the National Academies of Science, Engineering and Medicine, the National Association of Attorneys General, and the White House. In various ways, these organizations have recognized that acupuncture can play an important role in dealing with the opioid epidemic. This paper presents and analyzes the scientifific evidence supporting the effificacy of acupuncture in regard to opioid addiction.
Acupuncture Therapy ; Analgesics, Opioid ; adverse effects ; Clinical Trials as Topic ; Epidemics ; Humans ; Opioid-Related Disorders ; epidemiology ; therapy

The internationalization of Chinese acupuncture-moxibustion is inevitably accompanied by the localization of acupuncture-moxibustion. The localization of acupuncture-moxibustion will inevitably promote the diversified development of acupuncture-moxibustion technique and theory, which fully demonstrates the openness of modern acupuncture-moxibustion. In this study, the characteristics of localization of Chinese acupuncture-moxibustion in the United States are explored and studied from the following aspects: legal adjustment, technical adjustment, educational adjustment, service adjustment and theoretical adjustment.
Acupuncture Therapy ; Moxibustion ; United States

Although the foundations and evolution of Chinese medicine and Western medicine are very different, an increasing amount of research has revealed that those Eastern medicine principles practiced over thousands of years are confirmed by new technologies applied to the basic science of the human body. Recent scientific discoveries present enticing opportunities to reconcile Chinese medicine theories with Western biomedicine. Is there a trend toward the convergence of Eastern and Western medicine? Four studies which exemplify the potential for convergence are described in this article. The studies present findings in regard to mesentery, interstitium, a gut-lung axis, and lung-centered hematopoiesis, and were published recently in leading journals such as Science, Nature, and Lancet.
Hematopoiesis ; Humans ; Medicine, Chinese Traditional ; Meridians ; Organ Specificity

OBJECTIVETo observe the effect of matrix metalloproteinase-1 (MMP-1) from human host on degradation of dentin organic matrix of root dentin.

METHODSThe freshly extracted caries-free impacted teeth were selected. Teeth were cut transversely under the enamel-cementum junction into dentin sections with a thickness of about 5 mm. Then all sections with removal of cementum, pulp and predentin were randomly divided into four groups. In the first group, dentin sections were demineralized with acid solution for 21 days, and then incubated with MMP-1 solution for 7 days; the second group were only treated with acid solution for 21 days; the third group were only attacked by MMP-1 solution for 7 days; and the fourth group were untreated as a control. Then all sections were dehydrated in ascending strength of alcohol, critically dried, coated with platinum, and then observed under scanning electron microscope(SEM).

RESULTSThe dentin sections of root surface attacked by acid and MMP-1 showed that demineralization of dentin mineral and degradation of dentin matrix fibrae synchronously happened. The dentin matrix fibrae wasn't degradated in the groups treated with acid or MMP-1.

CONCLUSIONThe proteinases from human host may play an important role in the development of root surface caries. MMP-1 may distinctly degradate the organic matrix of demineralized dentin.

Dental Cementum ; Dental Enamel ; Dentin ; enzymology ; Humans ; Matrix Metalloproteinase 1 ; physiology ; Microscopy, Electrochemical, Scanning ; Root Caries ; enzymology ; Tooth Root ; enzymology

Objective: To explore the improvement influence of three combined exercise programs on sleep quality among college students with sleep disorders, so as to provide reference for the construction of exercise intervention programs for sleep disorder. Methods: A total of 35 college students with sleep disorders were randomly divided into AR group ( n =10), AM group ( n =8), RM group ( n =9) and CG group ( n =8). The three exercise groups were provided with exercise intervention for 8 weeks, 3 times/week , 60 min/time, and the CG group maintained previous lifestyle.Before and after the intervention,evaluation of subjective sleep quality and reduction rate of sleep disorder by PSQI, and Actigraph GT3X+ was used to monitor the changes in the objective sleep quality. Results: After the intervention,the total score of PSQI ( t =3.49, 2.31, 2.73), sleep quality score ( t =2.71, 3.00 , 5.29),sleep duration of AR group ( t =2.74), daytime dysfunction of AM group ( t =2.64) and sleep duration of RM group ( t = 2.29) significantly decreased ( P ＜0.05); The scores of sleep duration in AM group were significantly lower than those in AR group, and the scores of PSQI,sleep quality and daytime dysfunction in AM group were significantly lower than those in CG group (P<0.05). After intervention,TST ( t =-4.41, -8.37, -6.79) and SE ( t =-4.40, -5.86, -4.91) of AR group, AM group and RM group significantly increased( P <0.05), SOL ( t =4.18, 9.93), WASO ( t =2.91, 3.46) and NA ( t =4.80, 3.37) of AM group and RM group significantly decreased ( P <0.05). The changes of TST,SE and WASO in RM group were significantly higher than those in CG group, and SOL was significantly lower than those in CG group ( P <0.05). Conclusion The three kinds of combined exercise programs can improve the subjective and objective sleep quality of college students with sleep disorders, and reduce the incidence of sleep disorders; different combined exercise programs have different influence on the improvement of sleep quality of college students with sleep disorders, aerobic combined meditative movement exercise program has a prominent influence on the improvement of subjective sleep quality, and resistance combined meditative movement exercise program has a prominent influence on the improvement of objective sleep quality.

Objective:To explore the value of 70 keV virtual monoenergetic images(VMI) on dual-layer spectral detector CT in improving abdominal arterial phase images quality.Methods:The arterial phase images of patients who underwent abdominal enhanced scan on the Philips IQon dual-layer spectral detector CT in Shengjing Hospital of China Medical University from January to May 2019 were retrospectively analyzed. Fifty patients (12 males, 38 females) were included with age of 26-74 （53±12） years. Objective and subjective evaluation was performed in 120 kVp polychromatic conventional images (group CI) which derived from iterative reconstruction algorithm and group 70 keV VMI which derived from spectral reconstruction algorithm. The attenuation, noise, signal to noise ratio and contrast to noise ratio of abdominal aorta, celiac trunk, superior mesenteric artery, left kidney artery, right kidney artery, liver, spleen, pancreas, left and right kidney were compared between group 70 keV VMI and CI by using paired t test. Image quality of group 70 keV VMI and CI was evaluated by two radiologists independently with a 5-point scale and compared by Wilcoxon rank test. The inter-agreement of subjective scoring between the two radiologists was evaluated by Kappa test. Results:Except for pancreas, the attenuation of abdominal arteries and solid organs in group 70 keV VMI were higher than that of group CI and the difference was statistically significant ( Pall<0.05). The attenuation of pancreas was lower than that of group CI ( t=-3.097， P=0.003). The noise of abdominal arteries and solid organs in group 70 keV VMI showed lower values compared to group CI and the difference was statistically significant ( Pall<0.001). The signal to noise ratio as well as contrast to noise ratio of abdominal arteries and solid organs in group 70 keV VMI was higher than that of group CI and the difference was statistically significant ( Pall<0.001). Subjective image quality scores in group 70 keV VMI were higher than that of group CI and scores of group 70 keV VMI and CI were 5 (4,5) and 4 (4,5), respectively. The difference was statistically significant ( Z=-4.131, P<0.001). There was a good consistency of subjective image quality scores between two radiologists, which Kappa values of group 70 keV VMI and CI were 0.79 and 0.69, respectively. Conclusions:Compared to CI, 70 keV VMI derived from dual-layer spectral detector CT can optimize abdominal arterial phase image quality by decreasing the noise, improving the signal to noise ratio and contrast to noise ratio.

Objective:To investigate the value of programmed death-1（PD-1） expression on the T lymphocytes for the prognosis of septic patients.Methods:From September 2017 to May 2019, septic patients were included in Department of Intensive Care Unit at 6 hospitals. The PD-1 expression on T cells were measured by flow cytometry. Logistic regression was conducted to analyze independent risk factors related to death within 28 days，and receiver operating characteristic curve(ROC) was conducted to evaluate the prognostic value of PD-1 expression on T cells in septic patients.Results:A total of 64 septic patients were enrolled to this study，including 32 survivors and 32 deaths. The PD-1 expression on T cells in the death group was significantly higher than that in the surviving group ( P<0.05). Correlation analysis showed that the percentages of PD-1 +/CD3 +T cells and PD-1 +/CD8 +T cells were positively correlated with procalciton in ( r=0.313, P =0.015； r=0.375, P=0.003), logistic regression analysis showed that the percentages of PD-1 +/CD3 +，PD-1 +/CD4 +，PD-1 +/CD8 +T cells were independent risk factors for the death of sepsis patients. The percentage of PD-1 +/CD3 +T cell was 3.63%, with AUC 0.842, sensitivity to predict the mortality 96.43% and specificity 59.38%, ( P<0.000 1). The percentage of PD-1 +/CD4 +T cell was 4.65%, with AUC 0.847, sensitivity 96.43%, specificity 62.50%，( P<0.000 1). The percentage of PD-1 +/CD8 +T cell was 3.91%, with AUC 0.771, sensitivity 64.29%, specificity 81.25%，( P=0.000 3). Conclusions:The T cell PD-1 expression is an independent risk factor to predict the 28-day mortality in septic patients. Combining the proportions of PD-1 +/CD3 +, PD-1 +/CD4 +and PD-1 +/CD8 +T cells may further enhance the predictive value for death.

AIM To establish an HPLC-MS/MS method for the simultaneous content determination of pulegone,prim-O-glucosylcimifugin,5-O-methylvisammioside,columbianadin,saikosaponin a,saikosaponin d,ferulic acid,naringin,liquiritin and glycyrrhizic acid in Jingfang Granules.METHODS The analysis of methanol extract of this drug was performed on a 30℃thermostatic ZORBAX Eclipse Plus C18 column(2.1 mm×150 mm,1.8 μm),with the mobile phase comprising of methanol-0.1%formic acid flowing at 0.3 mL/min in a gradient elution manner,and electron spray ionization source was adopted in positive and negative ion scanning with multiple reaction monitoring mode.RESULTS Ten constituents showed good linear relationships within their own ranges(r>0.996 5),whose average recoveries were 96.7%-98.8%with the RSDs of 0.9%-1.9%.CONCLUSION This rapid,simple,sensitive and specific method can be used for the quality control of Jingfang Granules.