A methodology named immunoscorehas been proposed in recent years.It has been demonstrated to be a prognostic factor superior to the Union for International Cancer Control-American Joint Committee on Cancer (UICC-AJCC)TNMclassification.Over the past few years,it has gained a forefront posi-tion in colorectal cancer.It has the advantages of simple operation,low cost and high accuracy,and it is nee-ded for individual therapy.However,it still has its limitations.

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 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 In this study,we attempted to investigate whether Electroacupuncture(EA)could promote the autophagy function in macrophages of inflammatory skin tissues by activating CB2 receptor,thus relieving inflammatory pain induced by CFA in mice,and whether activation of CB2 receptor in NR8383 macrophages cell line can simulate the effect of EA on the autophagy function and mitochondrial damage.Methods Inflammatory pain model was induced by CFA injection into the planta the hind paw of wildtype and CB2 knockout mice.EA or sham EA was applied on the left Huantiao(GB30)and Yanglingquan(GB34)sites.Thermal hyperalgesia was determined with the Hargreaves test.Mechanical sensitivity was assessed with von Frey filaments.NR8383 microphage cell line was used to study the effect of CB2 activation on macrophage function induced by CFA.The expression level of autophagy protein LC3 and p62 in wildtype and CB2 knockout mice skin tissue and NR8383 cell line were determined by Western blot.And flow cytometry analysis was applied to detect damaged mitochondria and mitochondrial superoxide.Results CFA significantly reduced the thermal and mechanical pain threshold in both wildtype and CB2 knockout mice,comparing with the vehicle control groups(P<0.01).EA significantly inhibited thermal and mechanical hyperpathia induced by CFA in wildtype mice(P<0.05),but had no effect on CB2 knockout mice with CFA(P>0.05).CFA significantly increased the expression of p62 protein and decreased LC3-II/I ratio,which was inversed by EA in wildtype mice but wasn't affected by EA in CB2 knockout mice.CFA increased the expression of p62 protein and decreased LC3-II/I ratio in NR8383 cell line,which were inversed by CB2 agonist AM1241.CFA increased mitochondria damage,which were then attenuated by CB2 agonist AM1241.Conclusion The analgesic effect of EA on inflammatory pain induced by CFA was mediated by activation of CB2 receptor,which promoted the autophagy function and the clearance of damaged mitochondria in macrophage.

This study focused on various databases and literatures related to drug metabolism, collated and analyzed the information related to bacterial and human drug metabolic enzymes, and compared the similarities and differences between the information included in the database of bacterial and human drug metabolic enzymes. Results found more bacterial drug metabolic enzymes than human drug metabolic enzymes (9 703 vs 964), but much less than the total number of bacterial enzymes in BRENDA database (9 703 vs 20 835 235), indicating that the influence of bacteria on drug metabolism could have been greatly underestimated, and that further systematic research is needed.We summarized the progress and shortcomings of the current research on the influence of intestinal flora on drug metabolism, and proposed a research idea, that is, to predict through artificial intelligence whether intestinal bacterial proteins have the ability to metabolize drugs, to verify their biological functions by in vitro/in vivo experiments and gene editing, and to establish a database of drug metabolic enzymes from intestinal bacteria with complete annotation functions, in an attempt to provide a solid theoretical basis for further exploration of the effects of intestinal flora on drug metabolism.

Fengweiqi is the whole plant of Rhodiola dumulosa.It is a kind of natural and precious folk medicinal plant,mainly distributed on hillside rocks and crevasses at the altitude of 1 600-4 100 m.Fengweiqi mainly contains flavonoids,volatile oil,polysaccharides,various amino acids and trace elements.Modern pharmacological studies have shown that Fengweiqi has many significant pharmacological activities,such as anti-oxidation,anti-fatigue,anti-hypoxia and bacteriostasis.In this paper,the textual research,chemical constituents,pharmacological actions and artificial cultivation of Fengweiqi were reviewed in order to provide reference for further research and development of Fengweiqi resources.

Objective To explore the availability and safety of ileal ureter replacement combined with ileal augmentation cystoplasty in treating ureteral stenosis with contracted bladder.Methods From August 2015 to November 2018,three patients who underwent ileal ureter replacement combined with augmentation cystoplasty were treated with ileal ureter replacement combined with augmentation cystoplasty.There were 1 male and 2 females with the age ranging from 34 to 55 years (mean 39 years).Two patients suffered from left ureter stenosis,and one patient had stenosis on the both sides.The length of the ureter stenosis ranged from 6 to 18 cm (mean 9.8 cm).The preoperative bladder capacity ranged from 60 to 150 ml (mean 103.3 ml).In the surgery,part of ileum was used to replace the ureter,and the distal intestine was made into U-shape to enlarge the bladder.Results All operation were completed successfully.The operation time ranged from 220 to 400 min (mean 303.0 min),and the blood loss ranged from 150 to 500 ml (mean 283.3 ml).Laparoscopic surgery was performed in 1 case and open surgery in 2 cases.Three weeks after the surgery,the bladder volume underwent cystography ranged from 300 to 400 ml (mean 360.0 ml).Three months after the surgery,the postvoid residual urine volume ranged from 20 to 50 ml (mean 33.3 ml).Postoperative frequency and urgency of urine were completely relieved in 1 case,alleviated in 2 cases.Flank pain was completely relieved in 1 case,and alleviated in 2 cases postoperatively.The serum creatinine of 2 patients mildly increased after the surgery,while 1 patient remained stable.For complications,1 patient had urinary infection,and 1 patient suffered from metabolic acidosis.Conclusions Ileal ureteral replacement combined with augmentation cystoplasty can be the choice of treatment for long segment ureteral stenosis and enlarge the bladder simultaneously.The postoperative complications and the kidney functions should be regularly followed up.

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

Objective To investigate the quantity and function of CD8+T cells in peripheral blood of pa-tients with repeated implantation failure(RIF).Methods Thirty-seven patients with RIF and 19 healthy controls were enrolled in this study.The peripheral blood and endometrium were collected during the mid-luteal phase.The percentage of peripheral CD8+T subsets and the levels of perforin and granzyme B of peripheral CD8+T cells were determined by flow cytometry assay.The percentage of endometrial CD8+T cells was detected by IHC,the produc-tion of perforin and granzyme B of endometrial CD8+T cells was detected by IF. Results Compared with the con-trol group,the percentage of peripheral CD8+T cells in patients with RIF was not significantly changed(37.22% vs. 37.15%,P>0.05).However,the porportion of endometrial CD8+T cells in the RIF group was higher than that in the control group(1.99% vs.3.77%,P<0.001).The levels of perforin and granzyme B in peripheral blood and en-dometrial CD8+T cells in patients with RIF were similar with those in the control group.Conclusions Compared to the control group,the percentage of endometrial CD8+T was markedly upregulated in patients with RIF.However, the production of perforin and granzyme B were similar between the control group and the RIF group.

In clinical practice, red blood cell infusion needs to be based on the patient′s hemoglobin level. However, different guidelines recommend different thresholds for red blood cell infusion and the timing of blood transfusion initiation is still controversial due to the presence of these different thresholds. Meanwhile, the use of allogeneic blood products carries a certain risk of transfusion-related infections or organ damage. Therefore, initiating red blood cell infusion requires more evidence. This review discusses some new methods, namely central venous oxygen saturation, arterial venous oxygen difference, near-infrared spectroscopy, and perioperative transfusion trigger score. It aims to help evaluate blood transfusion trigger and provide reference for doctors when making transfusion decisions.