Extending the Grading of Recommendations Assessment, Development and Evaluation (GRADE) in Traditional Chinese Medicine (TCM): The GRADE-TCM.

AIM: To extend and form the "Grading of Recommendations Assessment, Development and Evaluation in Traditional Chinese Medicine" (GRADE-TCM). METHODS: Methodologies were systematically reviewed and analyzed concerning evidence-based TCM guidelines worldwide. A survey questionnaire was developed based on the literature review and open-end expert interviews. Then, we performed expert consensus, discussion meeting, opinion collection, external examination, and the GRADE-TCM was formed eventually. RESULTS: 265 Chinese and English TCM guidelines were included and analyzed. Five experts completed the open-end interviews. Ten methodological entries were summarized, screened and selected. One round of consensus was conducted, including a total of 22 experts and 220 valid questionnaire entries, concerning 1) selection of the GRADE, 2) GRADE-TCM upgrading criteria, 3) GRADE-TCM evaluation standard, 4) principles of consensus and recommendation, and 5) presentation of the GRADE-TCM and recommendation. Finally, consensus was reached on the above 10 entries, and the results were of high importance (with voting percentages ranging from 50 % to 81.82 % for "very important" rating) and strong reliability (with the Cr ranging from 0.93 to 0.99). Expert discussion meeting (with 40 experts), opinion collection (in two online platforms) and external examination (with 14 third-party experts) were conducted, and the GRADE-TCM was established eventually. CONCLUSION: GRADE-TCM provides a new extended evidence-based evaluation standard for TCM guidelines. In GRADE-TCM, international evidence-based norms, characteristics of TCM intervention, and inheritance of TCM culture were combined organically and followed. This is helpful for localization of the GRADE in TCM and internationalization of TCM guidelines.

Polymorphisms and AR: A Systematic Review and Meta-Analyses.

Background: Allergic rhinitis (AR) is an especially common disorder associated with both environmental and genetic factors, and a lot of researchers have attempted to find polymorphisms which predisposed to the disease. We conducted a meta-analysis of the most frequently researched polymorphisms to find those genes which may be susceptible to AR and then may be of value in diagnosis. Methods: Pubmed and China National Knowledge Infrastructure (CNKI) databases were searched to screen out eligible studies focusing on the correlation between polymorphisms and AR susceptibility, and then polymorphisms cited in at least 3 studies were selected. Results: The 142 papers originally selected cited 78 genes. Twelve genes (coinciding with 23 polymorphisms) were reported in more than three papers. Twenty-three polymorphisms were involved in the meta-analysis. Among the 23 polymorphisms, only 4 were found to be related to the risk of AR: IL-13 rs20541, CTLA-4 rs11571302, IL-4R RS1801275 and ACE (I/D). The remaining 19 of the 23 polymorphisms were not associated with AR. Conclusion: We found polymorphisms that could be used for AR diagnosing and those that were unrelated to AR. This may be the first step in detecting polymorphic combinations susceptible to AR (IL-13 RS20541, CTLA-4 RS11571302, IL-4R RS1801275 and ACE (I/D). In addition, our results may improve AR diagnosis and contribute to the intensive study of AR.

GRP78 mediates radiation resistance of a stem cell-like subpopulation within the MCF-7 breast cancer cell line.

Emerging evidence indicates that breast cancer-initiating cells (CICs) are relatively resistant to radiotherapy; however, the critical mechanisms determining breast CIC resistance to radiation remain elusive. In the present study, a subpopulation of cells displaying characteristics generally attributed to stem cells was identified within the breast cancer cell line MCF-7. This subpopulation displays cancer stem cell features characterized by overexpression of embryonic stem cell markers, high tumorigenic potential following transplantation into BALB/c-nu mice, self-renewal capacity and resistance to ionizing radiation (IR). Moreover, glucose­regulated protein 78KD (GRP78), which was found to play a crucial role in stem cell oncogenesis, was also shown to be overexpressed in this subpopulation. GRP78 is required for the cancer stem-like subpopulation cell resistance to IR, as knockdown of this gene augments the effects of IR, while overexpression of GRP78 increases the radiation resistance of the subpopulation to IR. These findings indicate that GRP78 acts as a potential therapeutic target aimed at tumor-generating subsets of breast cancer cells.

Mechanism underlying carbon tetrachloride-inhibited protein synthesis in liver.

AIM: To study the mechanism underlying carbon tetrachloride (CCl(4)) -induced alterations of protein synthesis in liver. METHODS: Male Sprague-Dawley rats were given CCl(4) (1 mL/100 g body weight) and (3)H-leucine incorporation. Malondialdehyde (MDA) level in the liver, in vitro response of hepatocyte nuclei nucleotide triphosphatase (NTPase) to free radicals, and nuclear export of total mRNA with 3'-poly A(+) were measured respectively. Survival response of HepG2 cells to CCl(4) treatment was assessed by methyl thiazolyl tetrazolium. Km and Vmax values of nuclear envelope NTPase activity in liver of rats treated with CCl(4) were assayed by a double-reciprocal plot. RESULTS: The protein synthesis was inhibited while the MDA level was significantly increased in liver of rats treated with CCl(4). In addition, CCl(4) decreased the NTPase binding capacity of nuclear envelope (Km value) in cultured HepG2 cells. Moreover, in vitro ferrous radicals from Fenton's system suppressed the NTPase activity of liver nuclear envelope in a dose-dependent manner. Down-regulation of the nuclear envelope NTPase activity indicated a lower energy provision for nucleocytoplasmic transport of mRNA molecules, an evidence in CCl(4)-treated HepG2 cells correspondingly supported by the nuclear sequestration of poly (A)(+) mRNA molecules in morphological hybridization research. CONCLUSION: Inhibition of mRNA transport, suggestive of decreased NTPase activity of the nuclear envelope, may be involved in carbon tetrachloride-inhibited protein synthesis in liver.

[Glucose regulated protein 78 kD].

One of the most important chaperones located on endoplasmic reticulum, GRP78, referred as BiP (immunoglobulin heavy chain binding protein), belongs to heat shock protein 70 family. GRP78 exists conservatively among a wide variety of biological species, and acts as a central regulator of endoplasmic reticulum (ER) functions, participating in ER protein folding and assembly process, and maintaining ER Ca2+ homeostasis, unfolded protein response and specific anti-apoptotic actions. Specific regulatory cis-elements such as ER stress response element (ERSE) and cAMP response element (CRE) were identified on the promoter of GRP78. Dynamic epigenetic interactions between specific transcription factors such as AFT6 and regulatory elements in GRP78 gene promoter might contribute to human GRP78 constitutive or inducible transcription, resulting from some physiological and pathological stresses. Recently, cellular relationship between GRP78 expression and hepatic steatosis, cancer and nervous system diseases in human was underwent further clinical and biochemical research, which will benefit to human beings.

Tunicamycin suppresses cisplatin-induced HepG2 cell apoptosis via enhancing p53 protein nuclear export.

Cisplatin is a widely used anticancer drug; however, resistance to cisplatin-based chemotherapy is a major cause of treatment failure in patients with tumors. The present study was undertaken to investigate whether and how endoplasmic reticulum (ER) stress initiated by tunicamycin, which inhibits glycosylation, influences cisplatin-induced apoptosis in HepG2 cells. Pretreatment of HepG2 cells with ER stress inducers brought about a decrease in both cisplatin-induced cytotoxic effects and apoptosis. In order to further explore the mechanism underlying tumor resistance to cisplatin, we observed that increased nuclear export of endogenous p53 protein by pharmacological inducers of ER stress, such as tunicamycin, was associated with the suppression of cisplatin-induced apoptosis. These results suggested that tumor suppressor p53 protein may play a key role in cisplatin-induced HepG2 cells apoptosis. It is therefore suggested that the treatment of some tumor patients with cisplatin be combined with the down-regulation of endogenous ER stress to improve the clinical results of cisplatin-based chemotherapy.

[Twin study on serum leptin and soluble leptin receptor in pubertal girls].

OBJECTIVE: To explore the changes of serum leptin, soluble leptin receptor (SOB-R) concentrations and free leptin index (FLI) throughout the puberty in females, and estimate the effects of genetic and environmental factors on these indices. METHODS: The population studied consists of 180 pairs of twins in 6 to 18 years old: 132 pairs of monozygotic twins and 48 pairs of dizygotic twins, who were all from Qingdao city, Shandong Province. Anthropometric and sexual characteristics were examined, fasting serum leptin and SOB-R contents were assayed by immunoradiometric assay and ELISA respectively. RESULTS: Serum leptin concentrations increased and SOB-R decreased throughout puberty (P < 0.05), brought out a sustained increase of FLI, especially in 7-9 and 12-14 years old (from 10.1 to 32.3 and 41.8 to 82.1 respectively); Leptin and FLI were positively, and SOB-R negatively correlated with morphological indices (correlation coefficients from 0.54 to 0.76, -0.23 to -0.42, respectively). Heritability of girl's serum leptin, SOB-R and FLI were 0.37, 0.84, 0.46 respectively. CONCLUSIONS: Serum free leptin index surge at 7-9 and 12-14 years old might be a predictor or trigger of the puberty onset and menarche, respectively; FLI might be determined by both genetic and environmental factors. Leptin should be substantially influenced by environment and reflect the fat mass of body compositions. SOB-R should be predominantly controlled by genetic factors yet. Genetic influence might be important in pubertal development and metabolic disorders.

[Twin study of free insulin-like growth factor-1 on female pubertal development].

OBJECTIVE: To explore the effect of insulin-like growth factor-1 (IGF-1) on female pubertal development and estimate heritability of the serum free IGF-1. METHODS: The study population consisted of 427 girls aged 6 to 18 years: 132 pairs monozygotic twins, 48 pairs same-sex dizygotic twins and 67 girls with opposite-sex twin, who were all from Qingdao city, Shan-dong province. Their breast development were examined by Tanner standard and fast serum free IGF-1 contents were assayed by immunoradiometric assay (IRMA). RESULTS: Serum free IGF-1 concentration increased firstly and decreased later throughout puberty. The peak of free IGF-1 was 12 years old, which equals to Tanner B2. Free IGF-1 heritability of group stage B1, B2-B5, pre-menarche and post-menarche were 0.53, 0.85, 0.48, 0.83 respectively, non-classed heritability was 0.53. CONCLUSION: Fast increase of serum free IGF-1 before 12 years might be a predictor of the growth spurt and breast development. Genetic effect on female serum free IGF-1 was significant in the process of puberty.

[Heritability analysis of insulin sensitivity and the effect of age and sex: a twin study in Chinese aged 5-18].

OBJECTIVE: To study the contribution of genes and environment to the variation of insulin sensitivity, and the effect of body mass index (BMI), age and sex. METHODS: A total of 296 healthy same-sex twin pairs were studied, including 223 monozygotic (MZ) and 73 dizygotic (DZ) twin pairs aged 5-18(mean age, 12+/-4 years). Microsatellite polymorphism (STR) was used to diagnose zygosity of twins. Model-fitting method using Mx package was performed to analyze the genetic and environmental variance of homeostasis model assessment insulin resistance index (HOMA IR) and HOMA beta cell function index (HBCI), before and after adjusting BMI. The effect of age and sex on the model was tested. RESULTS: HOMA IR was correlated with age, sex and BMI, while HBCI was correlated with age. Genetic analysis showed in ACE model, heritability of HOMA IR in boys and girls was different before and after BMI adjustment; heritability of HBCI was not different significantly. Heritability of HOMA IR and HBCI were 0.25, 0.24, respectively, after adjusting BMI. CONCLUSION: Gene and environment influence insulin sensitivity of children and adolescents together, whereas environment seems to play a more important role. The effect of BMI on estimating heritability of insulin sensitivity is considerable, differing from sex or age.