Objective To clone and express Dengue virus nonstructural protein 4A (NS4A) gene and express in eukaryotic cells.Then,to isolate and purify and isolate cellular proteins interacted with NS4A.Methods With specific primers,NS4A gene fragment tagged with FLAG and HA (FLAG-NS4A-HA) was amplified by PCR and cloned into an expression vector,pSG5 vector.Recombinant plasmid was transfected into A549 cells by LipofectAMINETM2000.Transient expression of FLAG-NS4A-HA was detected by Western blot.The NS4A interacting proteins were isolated and purified by tandem affinity purification (TAP) system using HA and FLAG antibodies,and then assayed by silver stained SDS-PAGE.Results Dengue virus NS4A gene tagged with FLAG and HA was successfully constructed into pSG5 vector and expressed in A.549 cells.Silver stained SDS-PAGE showed that the expressed NS4A and two potential interacting proteins that interact with NS4A were isolated after TAP purification and SDS-PAGE.Conclusion Cellular proteins that potentially interacted with Dengue virus NS4A were successfully purified and isolated,which provided a basis for further research.

Objective To establish a tandem affinity purification(TAP) system of innate immune-regulatory protein PKR and analyze PKR function, for the future screen and identification of novel PKR-interaction proteins. Methods PKR gene was amplified by PCR, and then cloned into a mammalian expression vector pcTAP-A. Recombinant pcTAP-PKR was transfected into PKR knock-down(PKRkd) HeLa cells by LipofectAMINE 2000,and the PKR overexpressed HeLa cells were harvested for mitogen-activated protein kinases(MAPK) activation analysis. Cell extracts of PKR overexpressed cells were purified using TAP kit and examined by Western blot. Results Cal modulin resin(CBP) and streptavidin resin(SBP) tagged PKR was detected in PKRkd HeLa cells as early as 24 h upon transfection with pcTAP-PKR, and its expression decreased at later time points. The overexpression of PKR was autophosphorylated, and thus involved in the regulation of MAPK actviation. After small-scale TAP kit purification, PKR protein was detectable by Western blot. Conclusion We have successfully established a TAP system that over-expresses functional PKR, providing a useful tool for the future study on the identification of PKR interacting proteins.

Objective:To analyze the effect of traditional Chinese medicine fasting therapy on atherogenic index of plasma and metabolic indices in patients with metabolic syndrome.Methods:A case-control study was conducted on 90 patients with metabolic syndrome who were treated at Wenzhou Traditional Chinese Medicine Hospital between June 2021 and June 2023. The patients were divided into two groups with 45 patients in each group using a random number table method. The control group was given a normal diet, while the observation group underwent traditional Chinese medicine fasting therapy. Blood lipid indexes, atherogenic index of plasma, Homeostasis Model Assessment of Insulin Resistance index, anthropometric indices, and oxidative stress response were compared between the two groups.Results:After treatment, the levels of low-density lipoprotein cholesterol, total cholesterol, and triglyceride in the observation group were (2.11 ± 0.26) mmol/L, (4.31 ± 0.26) mmol/L, and (1.39 ± 0.26) mmol/L, respectively. They were significantly lower than those in the control group [(2.95 ± 0.34) mmol/L, (5.24 ± 0.33) mmol/L, (2.68 ± 0.41) mmol/L, t = 13.16, 14.85, 17.82, all P < 0.05]. After treatment, the level of high-density lipoprotein cholesterol was significantly higher in the observation group than in the control group [(1.18 ± 0.09) mmol/L vs. (1.03 ± 0.04) mmol/L, t = 10.21, P < 0.001]. After treatment, atherogenic index of plasma and Homeostasis Model Assessment of Insulin Resistance index in the observation group were (0.10 ± 0.04) and (5.12 ± 0.42), respectively in the observation group. They were significantly lower than those in the control group [(0.28 ± 0.10), (5.80 ± 0.67), t = 11.21, 5.76, both P < 0.001]. After treatment, waist circumference, hip circumference, and visceral adiposity index in the observation group were (91.05 ± 4.26) cm, (98.16 ± 5.06) cm, and (3.94 ± 0.52), respectively. They were significantly lower than those in the control group [(95.55 ± 9.87) cm, (102.64 ± 9.84) cm, (5.66 ± 1.04), t = 2.80, 2.71, 9.92, all P < 0.05). After treatment, the level of superoxide dismutase in the observation group was significantly higher than that in the control group [(101.52 ± 13.52) U/mL vs. (80.01 ± 6.82) U/mL, t = 9.52, P < 0.001]. After treatment, the level of malondialdehyde in the observation group was significantly lower than that in the control group [(4.41±0.35) nmol/L vs. (6.26 ± 0.61) nmol/L, t = 17.64, P < 0.001). Conclusion:Traditional Chinese medicine fasting therapy can effectively reduce the atherogenic index of plasma, blood lipid level, waist circumference, hip circumference, and visceral adiposity index in patients with metabolic syndrome. It also reduces oxidative stress reactions and is highly effective.

OBJECTIVETo evaluate the roles of folic acid and beta-carotene in the chemoprevention of gastric and other gastrointestinal (GI) cancers.

METHODSIn a randomized, double-blind, placebo-controlled trial, a total of 216 patients with atrophic gastritis were randomly assigned to one of the four groups: (1) folate (FA, 20 mg per day plus vitamin B(12) 1 mg, intramuscularly, per month for one year, then 20 mg two times a week plus 1 mg per three months for the next year); (2) natural beta-carotene (N-betaC, 30 mg per day for first year, then 30 mg two times a week for the next); (3) synthetic beta-carotene (S-betaC, administered as in N-betaC); and (4) placebo. Follow-ups continued from 1994 to 2001.

RESULTSA total of 7 new cases of gastrointestinal cancers were diagnosed with 3 stomach, 1 colon and 1 esophageal cancers occurring in the placebo group; 1 stomach cancer in both of the N-betaC and S-betaC groups, and no cancer occurring in FA group. In terms of GI cancers, there was a significant reduction in the FA group, compared with the placebo group (P = 0.04). A similar trend was observed in both N-betaC and S-betaC groups (P = 0.07 - 0.08). Taken together, the three intervention groups displayed a highly significant decrease in occurrence (P = 0.004, vs placebo), and a lower risk for GI cancers (OR = 0.12; 95% confidence interval, 0.03 - 0.51). For development of gastric cancer, any one of the three active-treated groups did not reach statistically significant reduction. The FA group showed obvious improvement of the gastric mucosal lesions with more patients displaying lesions reversed or stable atrophy and inflammation (P = 0.04), reversed intestinal metaplasia (P = 0.06) at the end of follow-up, and reversed displasia (P = 0.017) at 12 months. Two cases of false jaundice were found in beta-carotene groups with no influence on administration, and no side-effects were reported in FA group.

CONCLUSIONSThis trial revealed the interventional effect of folic acid on the development of GI cancers, a similar effect of beta-carotene was also detected. Also, folic acid may be of use to treat atrophic gastritis by preventing or reversing the precancerous lesions.

Adult ; Aged ; Anticarcinogenic Agents ; therapeutic use ; Double-Blind Method ; Female ; Folic Acid ; adverse effects ; therapeutic use ; Gastric Mucosa ; pathology ; Gastrointestinal Neoplasms ; prevention & control ; Humans ; Male ; Middle Aged ; Patient Compliance ; Stomach Neoplasms ; prevention & control ; beta Carotene ; therapeutic use