Based on the concept of "blood exhaustion" from The Inner Canon of Yellow Emperor （《黄帝内经》）， a three-stage syndrome differentiation and treatment strategy for cancer-related anorexia-cachexia syndrome is proposeed. In the cancer-induced anorexia stage， the pathogenesis is characterized by cancer consuming the spleen and stomach， leading to stagnation of transportation and transformation in the middle jiao （焦）. Treatment should focus on strengthening the spleen， promoting appetite， dispersing accumulation， and aiding digestion， with modified Zisheng Pills （资生丸） in Extensive Notes on Medicine from Xian Xing Studio （《先醒斋医学广笔记》） or Zisheng Decoction （资生汤） in Records of Chinese Medicine with Reference to Western Medicine （《医学衷中参西录》）. In the pre-cachectic stage of malnutrition， the pathogenesis involves insufficient nourishment of blood and qi with essence depletion hindering production. Treatment should focus on nourishing blood and harmonizing ying （营）， warming yang and supplementing qi， and modified Huangqi Jianzhong Decoction （黄芪建中汤） can be used. In the cachectic stage， the pathogenesis involves blood deficiency and essence exhaustion， with blood stasis obstructing the collaterals. The therapeutic approach should focus on tonifying deficiency and replenishing essence， unblocking collaterals， and removing stasis， and modified Buzhong Yiqi Decoction （补中益气汤） and Zuo Gui Beverage （左归饮） are suggested.

This paper summarized professor WU Mianhua's experience in the treatment of tumors using Hehuanhua （Flos Albiziae）. He believes that the occurrence and development of tumors are closely related to emotions， and thereafter proposed the concept of "masses usually due to constraint， and should be treated by regulating the spirit".Hehuanhua is a light and aromatic flower that enters the heart， liver and spleen meridians， and is good at relieving constraint and regulating the spirit， harmonizing the center with aroma， which can be used to treat all kinds of tumor-related complications. Hehuanmi is flower buds， picked before the flowers fully blooms， have the effect of clearing and lightening upward， opening up the heart and mind， and unblocking liver qi. Commonly used formulas include self-made Ningxin Anmian Decoction （宁心安眠汤） to treat tumor-related insomnia， self-made Aitongxiao Formula （癌痛消方） to treat tumor-related pain， self-made Erhua Jieyu Formula （二花解郁方） to treat tumor-associated depression， and self-made Shugan Tiaohe Decoction （疏肝调和汤） to deal with peri-menopausal state after the treatment of tumors. Hehuanhua is an inflorescence， which captures the aromatic essence and has the function of removing the filth and turbidity， and can wake up the spleen and harmonize the stomach； therefore， it is often used in self-made Tiaomu Hezhong Decoction （调木和中汤） to treat chemotherapy-related digestive tract adverse reactions or combined with rose flowers to treat chemotherapy-induced oral ulcers.

Objective To evaluate the therapeutic effect of antibiotics combined with recombinant tuberculosis vaccine AEC/BC02 on Mycobacterium tuberculosis infection in guinea pigs. Methods Two weeks after guinea pigs were challenged subcutaneously with a high dose of Mycobacterium tuberculosis,the guinea pigs with the positive skin test responses to the recombinant ESAT6-CFP10 allergen were randomly di-vided into four groups:normal saline (NS),AEC/BC02,antibiotics and antibiotics+AEC/BC02. In antibiotics+AEC/BC02 group,guinea pigs firstly received isoniazid ( INH) and rifapentine ( RFT) treatment once a week for a total of three times,and then were immunized with a single dose of AEC/BC02 vaccine six times at 10-day intervals. Guinea pigs in AEC/BC02 and antibiotics groups were respectively vaccinated with AEC/BC02 vaccine and given INH and RFT treatment at the same dose and frequency as given to antibiotics+ AEC/BC02 group. Thirteen weeks after challenge,all guinea pigs were sacrificed for necropsy. Results The gross pathological scores of NS,AEC/BC02,antibiotics and antibiotics+AEC/BC02 groups were 83±8,77± 22,45±28 and 19±14,respectively. Antibiotics+AEC/BC02 group had a significantly lower gross pathological score than antibiotics,AEC/BC02 and NS groups (P<0. 05,P<0. 01,P<0. 01,respectively). Moreover,the gross pathological scores of antibiotics and AEC/BC02 groups were significantly decreased as compared with that of NS group (both P<0. 01). However,there was no significant difference between AEC/BC02 and NS groups. The spleen bacterial load of antibiotics+AEC/BC02 group was (2. 50±1. 26) lg CFU,which was sig-nificantly lower than those of NS [(4. 92+0. 52) lg CFU],AEC/BC02 [(4. 78+0. 84) lg CFU] and antibi-otics [(4. 39+0. 50) lg CFU] groups (P<0. 01). Compared with NS group,antibiotic and AEC/BC02 groups showed no significant difference in spleen bacterial load. Histopathological changes indicated different levels of granulomatous lesions appeared in lung tissues of all groups and the most severe change was ob-served in AEC/BC02 group,followed by that in NS,antibiotics and antibiotics+AEC/BC02 groups. Conclu-sion INH and RFT treatment in combination with AEC/BC02 vaccine in the treatment of guinea pigs with Mycobacterium tuberculosis infection was superior to either treatment alone as it significantly alleviated organ lesions and lowered the bacterial loads in spleen and lung.

More than 60% of active tuberculosis(TB) patients are smear- and culture-negative, constituting a prime group in the prevention and control of TB in China. In the existing laboratory testing technologies, immunological diagnosis is more advantageous than etiological diagnosis in the detection of smear-and culture-negative TB. Serum antibody detection reagents are cheap,easy to operate and time-sav-ing,and have been widely used in China. However,these agents are not stable in sensitivity and specificity, and because of that their accuracy in the diagnosis of smear-and culture-negative TB is doubtful. In this re-view,we summarize some problems in the use of serum antibody detection among smear- and culture-nega-tive pulmonary TB patients and discuss possible methods to solve these problems expecting to provide some ideas for promoting its development,application and policy formulation.

Objective To identify the cross-reactive antigens shared by Mycobacteria smegmatis(MS) and Mycobacteria tuberculosis(MTB) and to analyze their antigenicity.Methods Bacterial antigens were extracted from strains of MS and MTB by ultrasonication.Western blot assay was performed to analyze common antigens that reacted with both of the antiserum samples against MS and MTB.The extracted bacterial antigens were mixed with incomplete Freund′s adjuvant and then were injected into muscles of mice.Cytokines secreted by murine spleen lymphocytes following stimulation with various antigens of MS and MTB were determined by ELISPOT and flow cytometry on the 7th day.IgG levels in serum samples were detected by ELISA 7 days after injection.Results There were cross-reactive antigens shared by MS and MTB.Potent humoral immune responses and cellular immunity against both MS and MTB could be induced by those cross-reactive antigens after sensitization the mice by either MS or MTB antigens.Cytokines of IL-2 and IFN-γ in CD4+ and CD8+T cells of mice stimulated with MS or MTB antigens were significantly increased as compared with those of non-sensitization group and those of Brucella antigens stimulation group.ConclusionCross-reactive antigens shared by MS and MTS can effectively promote specific immune reactions to the infection of MTB, which provides a scientific basis for the development of tuberculosis vaccines.

RNA-binding protein exerts important biological function by specifically recognizing RNA motif. SELEX (Systematic evolution of ligands by exponential enrichment), an in vitro selection method, can obtain consensus motif with high-affinity and specificity for many target molecules from DNA or RNA libraries. Here, we combined SELEX with next-generation sequencing to study the protein-RNA interaction in vitro. A pool of RNAs with 20 bp random sequences were transcribed by T7 promoter, and target protein was inserted into plasmid containing SBP-tag, which can be captured by streptavidin beads. Through only one cycle, the specific RNA motif can be obtained, which dramatically improved the selection efficiency. Using this method, we found that human hnRNP A1 RRMs domain (UP1 domain) bound RNA motifs containing AGG and AG sequences. The EMSA experiment indicated that hnRNP A1 RRMs could bind the obtained RNA motif. Taken together, this method provides a rapid and effective method to study the RNA binding specificity of proteins.
Aptamers, Nucleotide ; Gene Library ; Heterogeneous Nuclear Ribonucleoprotein A1 ; genetics ; High-Throughput Nucleotide Sequencing ; Humans ; RNA ; SELEX Aptamer Technique

Objective To study the expressions of osteopontin (OPN),caspase-3 and mt-P53 proteins, and their relationship in gliomas. Methods Seventy gliomas specimens of patients (glioma group) were selected, and 10 samples of non-glioma brain tissue were used as control group. The SP method was used to detect the positive rates of protein expressions of OPN, caspase-3 and mt-P53 between two groups. The relationship between protein expressions of OPN, caspase-3 and mt-P53 in gliomas and grade of gliomas were detected by Western blot assay. Spearman rank correlation was compared between the positive expression of OPN, caspase-3 and rate mt-P53. Results The positive expression rates of OPN and mt-P53 were significantly higher in glioma group (64.29%and 60%) than those of control group (no positive expression), but the positive expression rate of caspase-3 was significantly lower than that of control group (47.14%vs. 90%, P<0.05). There were no significant differences in OPN, caspase-3 and mt-P53 expressions between different gender, age, tumor size and tumor position (P>0.05). The higher the WHO classification, the higher the positive expression rates of OPN and mt-P53 (P<0.001), and the lower the positive expression rate of caspase-3 (P<0.001). With the increased level of glioma grade, OPN and mt-P53 protein levels were increased, but caspase-3 protein expression level was decreased. There was a negatively correlation between OPN and the positive expression of caspase-3, but there was a positive correlation between OPN and the expression of mt-P53 (rs=-0.720 and 0.722, P<0.05). There was a negative correlation between caspase-3 and mt-P53 expressions (rs=-0.556, P<0.05). Conclusion The higher the WHO classification, the higher the positive expression rates of OPN and mt-P53, while the lower the positive expression rate of caspase-3. The study reveals that OPN, caspase-3 and mt-P53 expressions are associated with the occurrence and the progress of gliomas. The combined detection of them can contribute to the judgment of biological behavior of gliomas.

Aim To establish and optimize a method for screening HIV-1 integrase 3′-processing inhibitor. Methods Fluorescence resonance energy transfer ( FRET) was used to create an assay for screening in-tegrase 3′-processing inhibitors; wavelength was de-fined by DNaseⅠ; factors affecting IN activity were optimized, including buffer composition, substrate con-centration, enzyme concentration, metal ion concentra-tion. Results Integrase 3′-processing optimizing reac-tion conditions were buffer 1 , 500 nmol · L-1 sub-strate, 1 μmol·L-1 integrase, 20mmol·L-1 magne-sium ion. Positive drug raltegravir and myricetin could effectively inhibit integrase 3′-processing activity using this assay. Two integrase 3′-processing inhibitors were screened by this method. Conclusion The method for screening HIV-1 integrase 3′-processing inhibitor is successfully established and optimized.

Objective To assess the efficacy of injection of sodium hyaluronate in the treatment of patients with disc perforation by using cone-beam computed tomography (CBCT) and temporomandibular joint (TMJ) function.Methods Twenty-eight patients (38 joints) with disc perforation observed through arthrography who received injection of sodium hyaluronate and other 20 patients (29 joints) who received disc repair were included in this study.Pain (visual analog scale,scores 0 to 100),maximal mouth opening,modified Helkimo's clinical dysfunction index and TMJ examinations with CBCT were obtained before and 3,9 months after treatment.Results A statistically significant increase (P＜0.05) in TMJ clinical outcome was observed at each follow-up.The maximal mouth opening (MMO) increased 5.89 mm and 7.64 mm at 3 and 9 months follow-up in the hyaluronic acid (HA) group,and the MMO increased 2.75 mm and 6.10 mm at 3 and 9 months follow-up in the operation group.Osseous remodeling occurred in 66％ (25 of 38 joints) of patients after HA injection and 55％(16 of 29 joints) of patients appeared bone remodeling after disc repair.Conclusions Both injection of hyaluronic acid and disc repair are effective for treatment of temporomandibular joint disc perforation.

Objective To comparatively evaluate the effects of a recombinant Mtb protein ESAT 6-CFP10 ( rESAT6-CFP10 ) and a purified protein derivative ( PPD ) as skin test reagents in guinea pigs . Methods Guinea pigs were sensitized with different Mycobacteria species .After sensitization , all guinea pigs were intradermally injected with rESAT6-CFP10 and PPD.At 48 h after the injection, the size of ery-thema at injection sites was measured by using a double-blind method .For guinea pigs sensitized with viable Mtb, the size of erythema at injection sites were measured at 24 h after the injection .The positive conversion rates of skin test with rESAT 6-CFP10 and PPD were calculated .Results The results of PPD skin test were positive in all guinea pigs sensitized with viable Mtb , killed Mtb and BCG with erythema diameters of (11.4 ±0.9) mm, (11.8±1.1) mm and (13.2±0.8) mm, respectively.Positive skin test with rESAT6-CFP10 was only observed in guinea pigs infected by viable Mtb-showing erythema diameters of (13.7±5.7) mm. The skin test with rESAT6-CFP10 was negative in guinea pigs sensitized by killed Mtb-and vaccinated by BCG.The skin tests by using rESAT6-CFP10 and PPD were performed on randomly selected guinea pigs at ninth day after infection by Mycobacterium tuberculosis H37Rv.At the 2nd week, totally 24 selected guinea pigs showed positive skin test results with rESAT6-CFP10 (24/24) with erythema diameters of (19.9± 3.0) mm, while only 15 out of 24 had positive PPD skin test with erythema diameters of (6.1±5.5) mm. At the 4th week, all guinea pigs showed positive PPD skin test (3/3) with erythema diameters of (12.7± 2.5) mm.Conclusion The skin test by using recombinant ESAT 6-CFP10 protein can effectively distin-guish viable Mtb infection from BCG vaccination and killed Mtb sensitization , which is a more suitable anti-gen than PPD for the early diagnosis of Mtb infection .