American medical insurance is mainly composed of public medical insurance borne by the government and private medical insurance. The main cause of death in this country are ischemic heart disease, Alzheimer’s disease, and lung cancer. The motality rate mainly caused by drug use disorders, chronic kidney disease and Alzheimer’s disease. The development of Chinese herbal medicine is relatively slow and difficult. The education system has covered Traditional Chinese Medicine (TCM). However, there still exist certain challenges of the development of TCM acupuncture faces the challenge of localization; Chinese herbal medicine still lacks standardization and TCM education needs to be standardized. Based on the current situation, it is suggested to focus on the development of acupuncture and moxibustion, to promote the registration and declaration of Chinese herbal medicine products for treating difficult diseases, and to strengthen the international exchange of TCM education, so as to promote the development and spread of TCM in the United States.

Traditional T vector cloning method requires onerous procedures for identifying recombinant, and directional cloning was impossible. In order to overcome these problems, we have devised a directional T vector pETG based on pET-23a(+). For gene cloning, 7 bp partial LacO sequence was introduced into DNA fragment to reconstitute a full length LacO with Bfu I digested T vector. After transformation, blue colonies were selected on LB plate supplemented with X-gal. Restriction enzyme digestion and PCR identification showed that all blue colonies contained the directionally inserted recombinants and the recombinant efficiency was nearly 100%. We have successfully cloned 103 genes from human liver cDNA; in the study complicated procedures for screening of recombinant were not required. Eight pETG clones were picked for protein expression, and all the clones successfully produced corresponding proteins. We demonstrated that the directional T vector was successfully constructed, and it was very suitable for gene cloning and expression.
Cloning, Molecular ; methods ; DNA, Complementary ; biosynthesis ; genetics ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; genetics ; Genetic Vectors ; genetics ; Humans ; Liver ; chemistry ; Polymerase Chain Reaction ; methods ; Recombinant Proteins ; biosynthesis ; genetics

Objective:To establish a predictive model of moderate to severe pain in patients with hepatocellular carcinoma (HCC) undergoing transcatheter arterial chemoembolization (TACE).Methods:264 patients with HCC who underwent TACE operation in Southern Medical University from January 2017 to April 2018 were selected as the modeling set. The pain was assessed by numeric rating scales. The patients were divided into pain group ( n=96) and non-pain group ( n=168) according to whether moderate to severe pain occurred within 24 hours after the operation. Binary Logistic regression analysis were performed for variables that were statistically significant in the univariate analyses. The predictive nomogram was constructed and the internal validation was performed. In addition, 87 patients with HCC who underwent TACE operation from January 2020 to June 2020 were selected as the validation set for external validation. Results:In the modeling set, 96 patients (36.36%) had moderate to severe pain within 24 hours after TACE operation in 264 patients with HCC, and the dosage of morphine intramuscularly injected within 24 hours was 1015 mg, with an average of 10.57 mg per patient. Multivariate Logistic regression analysis showed that preoperative pain, the distance between the tumor and capsule ≤2 cm, high prothrombin activity, dosage of lipiodol>10 ml, and several thromboembolic tumors were independent risk factors for moderate to severe pain after TACE ( P<0.05). Age>50 was the protective factor of moderate to severe pain after TACE ( P<0.05). The area under ROC curve was 0.799 (95% CI: 0.745-0.853) in the modeling set. The area under Roc curve for internal validation and external validation were 0.780 and 0.788, respectively. The calibration curves showed satisfactory agreements between the model predicted probability and the actually observed probability. Conclusion:The predictive model of moderate to severe pain after TACE was established in this study has good differentiation and accuracy, it has certain guiding significance for predicting the high-risk group of moderate to severe pain after TACE operation and formulating the targeted prevention strategy.

Background: Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disorder mainly caused by homozygous deletions that include exon 7 of the survival motor neuron 1 (SMN1) gene. A nearby paralog gene, SMN2, obstructs the specific detection of SMN1. We optimized a duplexed real-time PCR approach using locked nucleic acid (LNA)-modified primers to specifically detect SMN1. Methods: An LNA-modified primer pair with 3´ ends targeting SMN1 specific sites c.835-44g and c.840C was designed, and its specificity was examined by real-time PCR and Sanger Sequencing. A duplexed real-time PCR approach for amplifying SMN1 and control gene albumin (ALB) was developed. A randomized double-blind trial with 97 fresh peripheral blood samples and 25 dried blood spots (DBS) was conducted to evaluate the clinical efficacy of the duplexed approach. This new approach was then used to screen 753 newborn DBS. Results: The LNA-modified primers exhibited enhanced specificity and 6.8% increased efficiency for SMN1 amplification, compared with conventional primers. After stabilizing the SMN1 test by optimizing the duplexed real-time PCR approach, a clinical trial validated that the sensitivity and specificity of our new approach for detecting SMA patients and carriers was 100%. Using this new approach, 15 of the screened 753 newborns were identified as carriers via DBS, while the rest were identified as normal individuals. These data reveal a carrier rate of 1.99% in Hunan province, South Central China. Conclusions We have developed a novel, specific SMN1 detection approach utilizing real-time PCR with LNA-modified primers, which could be applied to both prenatal carrier and newborn screening.