Guideline on Establishing Diagnostic Criteria for Chinese Medicine Syndromes.

AIM: To formulate the guideline for the development of diagnostic criteria for Chinese medicine syndromes, which can contribute to standardization of development of Chinese medicine syndrome diagnostic standards. METHODS: We embark into account on the development of Guideline on Establishing Diagnostic Criteria for Chinese Medicine Syndromes through Delphi method with reference to the existing technical system of diagnostic criteria for Chinese medicine syndromes and relevant criteria. RESULTS: Our guideline specifies principles, methods, and procedures for the formulation of diagnostic criteria for Chinese medicine syndromes. CONCLUSIONS: It is a comprehensive and systematic evidence-based guideline, and we hope this guideline can be applied as a reference in developing diagnostic criteria for Chinese medicine syndromes in other disciplines. It is also applicable to the formulation of diagnostic criteria for relevant clinical, educational, and scientific research by hospitals, institutes, and academies.

[Serum metabolomics of chronic obstructive pulmonary disease with lung-Qi deficiency syndrome].

The present study analyzed the potential biomarkers of chronic obstructive pulmonary disease(COPD) with lung-Qi deficiency syndrome by non-targeted metabolomics and explored the biological basis of this syndrome. Blood samples of 96 COPD patients with lung-Qi deficiency syndrome(COPD with lung-Qi deficiency syndrome group) and 106 healthy people(healthy control group) were collected, and the metabolic profiles of both groups were analyzed by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS). Multivariate statistical analysis and differential metabolite screening were carried out by using Progenesis QI and Simca-P. Metabolic pathways were constructed through the MetaboAnalyst. Seven potential biomarkers, such as L-cystathionine, protoporphyrinogen Ⅸ, and citalopram aldehyde, were identified. Compared with the results in the healthy control group, the content of citalopram aldehyde, N1-methyl-2-pyridone-5-carboxamide, and 11ß,17ß-dihydroxy-4-androsten-3-one was significantly up-regulated, while that of the other four compounds such as L-cystathionine, dihydrotestosterone, protoporphyrinogen Ⅸ, and D-urobilinogen was down-regulated. These potential biomarkers involved six metabolic pathways, including cysteine and methionine metabolism, porphyrin and chlorophyll metabolism, drug metabolism of cytochrome P450, steroid hormone biosynthesis, glycine, serine, and threonine metabolism, and nicotinate and nicotinamide meta-bolism. This study is expected to provide a certain scientific basis for the research on traditional Chinese medicine syndrome of COPD with lung-Qi deficiency syndrome from the molecular biology level.

Oxygen-Delivering Polyfluorocarbon Nanovehicles Improve Tumor Oxygenation and Potentiate Photodynamic-Mediated Antitumor Immunity.

Hypoxia is a critical cause of tumor immunosuppression, and it significantly limits the efficacy of many anticancer modalities. Herein, we report an amphiphilic F11-derivative-based oxygen-delivering polyfluorocarbon nanovehicle loading photodynamic DiIC18(5) and reactive oxygen species (ROS)-sensitive prodrug of chemo-immunomodulatory gemcitabine (PF11DG), aimed at relieving tumor hypoxia and boosting antitumor immunity for cancer therapy. We optimized F11-based polyfluorocarbon nanovehicles with a 10-fold enhancement of tumor oxygenation. PF11DG exhibited intriguing capabilities, such as oxygen-dissolving, ROS production, and responsive drug release. In tumors, PF11DG exhibited flexible intratumoral permeation and boosted robust antitumor immune responses upon laser irradiation. Notably, the treatment of PF11DG plus laser irradiation (PF11DG+L) significantly retarded the tumor growth with an 82.96% inhibition in the 4T1 breast cancer model and a 93.6% inhibition in the PANC02 pancreatic cancer model with better therapeutic benefits than non-oxygen-delivering nanovehicles. Therefore, this study presents an encouraging polyfluorocarbon nanovehicle with deep tumor-penetrating and hypoxia-relieving capacity to boost antitumor immunity for cancer treatment.

Tumor-permeated bioinspired theranostic nanovehicle remodels tumor immunosuppression for cancer therapy.

The robust immunosuppressive microenvironment in tumor represents a key challenge of cancer treatment, and their modulations by versatile therapeutic agents are critically hampered by the limited intratumoral delivery. Herein, we report a bioinspired tumor-responsive theranostic nanovehicle (BTN) with striking tumor-penetrating capability to relieve the profound immunosuppression in tumor for effective cancer therapy. BTN is designed by loading tumor-activated melittin pro-peptide, theranostic photochlor and reactive oxygen species (ROS)-responsive prodrug of chemo-immunomodulator gemcitabine into a bioinspired lipoprotein-based nanovehicle, which display prominent tumor accumulation and flexible intratumoral permeation. Notably, the BTN-mediated combinational treatment caused drastic elimination of multiple immunosuppressive cells and remarkable infiltration of cytotoxic lymphocytes in tumor, thereby essentially relieving the tumor immunosuppression and strikingly depressing the tumor growth. Therefore, this design provides an encouraging delivery nanoplatform with distinguished immunosuppression-relieving capacity for effective cancer therapy.

LncRNA TSLNC8 synergizes with EGFR inhibitor osimertinib to inhibit lung cancer tumorigenesis by blocking the EGFR-STAT3 pathway.

The roles of lncRNA TSLNC8 and its synergetic effects with osimertinib remain unknown in lung cancer. qRT-PCR or western blotting was performed to determine the expression levels of TSLNC8, EGFR and STAT3. Colony formation and MTT assays were used to evaluate cell proliferation. Transwell and wound healing assays were performed to assess migration and invasion abilities. Flow cytometry with Annexin V/PI staining was used to detect changes in cell apoptosis. Nude mice subcutaneous tumor model was constructed and used for validating the effects of TSLNC8 and osimertinib in vivo. Expression of TSLNC8 was down-regulated in clinical lung cancer tissues and cell lines. TSLNC8 overexpression or osimertinib administration led to promotion of apoptosis and inhibition of cell proliferation, migration and invasion, as well as deactivation of the EGFR-STAT3 pathway, whereas TSLNC8 knockdown had opposite effects. Moreover, the above effects of osimertinib were remarkably enhanced by TSLNC8 overexpression and inhibited by TSLNC8 knockdown, respectively. Meanwhile, the effects of TSLNC8 overexpression were reversed by STAT3 activation or EGFR overexpression. In the animal model, combination of TSLNC8 overexpression and osimertinib administration resulted in efficient suppression of tumor growth. In this study, we revealed a TSLNC8-EGFR-STAT3 signaling axis in lung cancer, and TSLNC8 overexpression significantly enhanced the anti-tumor effects of osimertinib via inhibiting EGFR-STAT3 signaling.

Targeting peptide-decorated biomimetic lipoproteins improve deep penetration and cancer cells accessibility in solid tumor.

The limited penetration of nanoparticles and their poor accessibility to cancer cell fractions in tumor remain essential challenges for effective anticancer therapy. Herein, we designed a targeting peptide-decorated biomimetic lipoprotein (termed as BL-RD) to enable their deep penetration and efficient accessibility to cancer cell fractions in a tumor, thereby improving the combinational chemo-photodynamic therapy of triple negative breast cancer. BL-RD was composed of phospholipids, apolipoprotein A1 mimetic peptide (PK22), targeting peptide-conjugated cytotoxic mertansine (RM) and photodynamic agents of DiIC18(5) (DiD). The counterpart biomimetic lipoprotein system without RM (termed as BL-D) was fabricated as control. Both BL-D and BL-RD were nanometer-sized particles with a mean diameter of less than 30 nm and could be efficiently internalized by cancer cells. After intravenous injection, they can be specifically accumulated at tumor sites. When comparing to the counterpart BL-D, BL-RD displayed superior capability to permeate across the tumor mass, extravasate from tumor vasculature to distant regions and efficiently access the cancer cell fractions in a solid tumor, thus producing noticeable depression of the tumor growth. Taken together, BL-RD can be a promising delivery nanoplatform with prominent tumor-penetrating and cancer cells-accessing capability for effective tumor therapy.

Bioinspired lipoproteins-mediated photothermia remodels tumor stroma to improve cancer cell accessibility of second nanoparticles.

The tumor stromal microenvironments (TSM) including stromal cells and extracellular matrix (ECM) form an abominable barrier hampering nanoparticles accessibility to cancer cells, significantly compromising their antitumor effects. Herein, we report a bioinspired lipoprotein (bLP) that can induce efficient photothermia to remodel TSM and improve second bLP accessibility to cancer cells for antitumor therapy. The multiple stromal cells and ECM components in TSM are remarkably disrupted by bLP-mediated photothermal effects, which cause a 4.27-fold enhancement of second bLP accumulation in tumor, deep penetration in whole tumor mass and 27.0-fold increase of accessibility to cancer cells. Of note, this bLP-mediated TSM-remodeling to enhance cancer cell accessibility (TECA) strategy produces an eminent suppression of tumor growth and results in a 97.4% inhibition of lung metastasis, which is superior to the counterpart liposomes. The bLP-mediated TECA strategy provides deeper insights into enhancing nanoparticle accessibility to cancer cells for antitumor therapy.

Deep Tumor-Penetrated Nanocages Improve Accessibility to Cancer Stem Cells for Photothermal-Chemotherapy of Breast Cancer Metastasis.

Cancer stem cells (CSCs) are proposed to account for the initiation of cancer metastasis, but their accessibility remains a great challenge. This study reports deep tumor-penetrated biomimetic nanocages to augment the accessibility to CSCs fractions in tumor for anti-metastasis therapy. The nanocages can load photothermal agent of 1,1-dioctadecyl-3,3,3,3-tetramethylindotricarbocyanine iodide (DBN) and chemotherapeutic epirubicin (EBN) to eradicate CSCs for photothermal-chemotherapy of breast cancer metastasis. In metastatic 4T1-indcued tumor model, both DBN and EBN can efficiently accumulate in tumor sites and feasibly permeate throughout the tumor mass. These biomimetic nanosystems can be preferentially internalized by cancer cells and effectively accessed to CSCs fractions in tumor. The DBN+laser/EBN treatment produces considerable depression of primary tumor growth, drastically eradicates around 80% of CSCs fractions in primary tumor, and results in 95.2% inhibition of lung metastasis. Thus, the biomimetic nanocages can be a promising delivery nanovehicle with preferential CSCs-accessibility for effective anti-metastasis therapy.

Apoferritin nanocages loading mertansine enable effective eradiation of cancer stem-like cells in vitro.

Cancer stem-like cells (CSCs) are proposed to be responsible for tumor metastasis, resistance and relapse after therapy, but are unable to be eliminated by many current therapies. Herein, we report that the apoferritin nanocages loading cytotoxic mertansine (M-AFN) can significantly improve their uptake in CSCs-enriched tumorspheres and effectively eradicate CSCs in tumorspheres for anticancer therapy. M-AFN were uniformly nanocage structures with the mean diameter of 11.26 ±â€¯2.58 nm and the loading capacity of 0.62%. In the CSCs-enriched tumorsphere model, M-AFN could be preferentially internalized by tumorsphere cells and the average half-inhibitory concentration (IC50) of M-AFN was obviously reduced by 5.46-fold when comparing to the parent 4T1 breast cancer cells. Moreover, both the already existing tumorspheres and the formation of secondary tumorspheres were drastically disrupted by M-AFN, but barely impacted by mertansine alone. The flow cytometer analysis showed the CSCs fractions in tumorspheres were considerably reduced by the M-AFN treatment. Therefore, the apoferritin nanocages represent an encouraging nanoplatform to eradicate CSCs for effective anticancer therapy.

Cell-penetrating peptide-based nanovehicles potentiate lymph metastasis targeting and deep penetration for anti-metastasis therapy.

Lymph metastasis is a vital pathway of cancer cell dissemination, and insidious lymph node metastasis increases the risk of distant cancer metastasis. Current therapies for lymph metastasis are largely restricted by limited targeting and penetration capacity. Herein, we report that an r9 cell-penetrating peptide-based cabazitaxel nanovehicle (r9-CN) displays prominent lymph metastasis targeting and deep penetration ability after intravenous injection for effective anti-metastasis therapy. Methods: The r9-CN and CN nanovehicles were prepared by thin film dispersion, using DSPE-PEG2000 as the nano-carrier material and cabazitaxel as the model drug to fabricate r9-modified nano-micelles by self-assembly. The morphology, size, and stability in physiological solutions of r9-CN and CN were characterized. The targeting, biodistribution, deep penetration, and therapeutic efficacy of r9-CN and CN were systematically explored in vitro and in vivo. Results: The r9-CN nanovehicle consists of homogeneous particles with a mean diameter of 13 nm and zeta potential of +0.75 mV. Compared with the nanovehicle lacking the r9 peptide (CN), r9-CN exhibits long retention and deep penetration in the tumor mass, and considerably enhances accumulation and flexible permeation in metastatic lymph nodes, thereby notably suppressing primary tumor growth, lymph node metastasis, and distant lung metastasis. Conclusion: The cumulative findings reveal that r9-CN offers a promising delivery platform, enabling efficient lymph metastasis targeting and deep penetration for effective anti-metastasis therapy.

Bioengineered Macrophages Can Responsively Transform into Nanovesicles To Target Lung Metastasis.

Specific drug delivery to metastatic tumors remains a great challenge for antimetastasis therapy. We herein report a bioengineered macrophage-based delivery system (LD-MDS) that can be preferentially delivered to lung metastases and intelligently transformed into nanovesicles and secondary nanovesicles for antimetastasis therapy. LD-MDS was prepared by anchoring a legumain-specific propeptide of melittin (legM) and cytotoxic soravtansine (DM4) prodrug onto the membrane of living macrophages. LD-MDS is responsively activated by legumain protease and converted into DM4-loaded exosome-like nanovesicles (DENs), facilitating efficient internalization by metastatic 4T1 cancer cells and considerable cell death. Afterward, the damaged 4T1 cells can release secondary nanovesicles and free drug molecules to destroy neighboring cancer cells. In vivo, LD-MDS displays superior targeting efficiency for lung metastatic lesions with diameters less than 100 µm and remarkably inhibits lung metastasis. This study provides a new opportunity to explore endogenous macrophages as living drug delivery vehicles with controlled drug release to target metastatic lung tumors.

Traditional Chinese medicine ZHENG identification of bronchial asthma: Clinical investigation of 2500 adult cases.

OBJECTIVES: This study aims to employ more comprehensive approaches to analyze the primary and secondary symptoms of clinical common TCM ZHENG of bronchial asthma according to the clinical investigation of 2500 adult cases. METTHODS: Patients met the inclusion criteria were surveyed by associate chief physician or chief physician through the TCM Clinical investigation questionnaire containing general demographic information, signs and symptoms, tongue and pulse conditions. Firstly, absolute frequency, cumulative frequency, Chi-squared test were adopted to reflect the clinical common ZHENG in three asthma stages, then the logistic regression analyses, the frequency methods were combined to distinguish the primary and secondary symptoms of the common ZHENG. RESULTS: Of the 2500 questionnaires, 2428 valid questionnaires were got, with the number in acute exacerbation stage was 1273, and that 586 in chronic persistent stage, and 569 in clinical remission stage, in which the number of excess syndromes, the deficiency-excess complex syndromes and deficiency syndromes corresponding to the above three stage respectively accounted on 55.7%, 69.97%, and 76.50%. According to the distribution of each ZHENG, ten clinical common ZHENG were distinguished by criteria of the frequency percent value at least 10.0% and cumulative percent value reach to 70% or above. Then based on the OR and frequency value of each symptom, the primary symptoms were tentative identified by OR value≧3 with percent value≧50%, and that of the secondary symptoms were OR value between 1 and 3 with percent value between 25% and 50%. CONCLUSIONS: Ten common ZHENG of asthma with each primary and secondary symptoms in three stages are identified through the clinical investigation: Exterior cold with interior fluid retention, Phlegm-turbidity obstructing the lung, Phlegm-heat obstructing the lung, Wind-phlegm obstructing the lung, Blood stasis, Qi deficiency of the lung, Qi deficiency of the lung and spleen, Qi deficiency of the lung and kidney, Qi and Yin deficiency of the lung and kidney, Yang deficiency of the lung and kidney.

Functional and psychosocial effects of pulmonary Daoyin on patients with COPD in China: study protocol of a multicenter randomized controlled trial.

BACKGROUND: Chronic obstructive pulmonary disease (COPD) is a major public health problem worldwide. Pulmonary rehabilitation (PR) is an established intervention for the management of patients with COPD. Exercise training is an important part of PR, and its effectiveness in patients with COPD is well established. However, alternative methods of PR training such as Daoyin have not been appropriately studied. Hence, alternative forms of exercise training that require less exercise equipment and no specific training place should be evaluated. This paper describes the study protocol of a clinical trial that aims to determine if pulmonary Daoyin training will improve the exercise capacity and psychosocial function of patients with COPD in China. METHODS AND DESIGN: A multicenter, randomized, controlled trial will be conducted. A total of 464 patients meeting the inclusion criteria will be enrolled into this study with 232 patients in each of the trial group and the control group. Based on patient education, patients in the trial group will receive pulmonary Daoyin and continue with their usual therapy for three months. In the control group, patients will continue with their usual therapy. The primary outcome measures are exercise capacity assessed by the six-minute walking distance test and lung function. Secondary outcomes include dyspnea and quality of life. Measurements will be taken at baseline (month 0) and after the study period (month 3). DISCUSSION: It is hypothesized that pulmonary Daoyin will have beneficial effects in improving exercise capacity and psychosocial function of patients with stable COPD, and will provide an alternative form of exercise training that is accessible for the large number of people with COPD. TRIAL REGISTRATION: This trial has been registered in ClinicalTrials.gov. The identifier is NCT01482000.

[An expert analysis of diagnostic criteria for traditional Chinese medicine syndromes of chronic obstructive pulmonary disease at acute exacerbation stage].

OBJECTIVE: To investigate the common syndromes, clinical symptoms and characteristics of chronic obstructive pulmonary disease at acute exacerbation stage (AECOPD). METHODS: Delphi method was used to organize the AECOPD expert questionnaire. This questionnaire was distributed to 30 selected experts of respiratory diseases. The resulting data were statistically analyzed by Chi-square test and statistical description such as mean, coefficient of variation and ratio of full marks. Common syndrome: mean≥3.00, coefficient of variation <30% and ratio of full marks≥10% and common clinical symptoms: for major symptoms present, the index mean≥4.00 and coefficient of variation <30% and for minor symptom, index mean≥3.00 and coefficient of variation <30% were used as criteria. RESULTS: Twenty-nine valid expert questionnaires were received. The coefficient of positivity was 96.67%, of authority was 0.835 and of coordination equals 0.359 (χ(2)=463.15, P=0.001). The mean, coefficient of variation and ratio of full marks for AECOPD syndrome of wind and cold invasion of lung were 4.12, 21.50%, and 34.48%, respectively. Those for syndrome of exogenous cold-evil and fluid-retention were 4.39, 14.32%, 48.28%; those for syndrome of wind and heat invasion of lung were 3.54, 30.77%, 6.90%;those for syndrome of phlegm-heat obstruction of the lung were 4.85, 9.23%, 89.66%; those for syndrome of pulmonary stagnation of phlegm were 4.36, 15.57%, 48.28%; those for syndrome of external cold and internal heat were 4.59, 16.27%, 65.52%; those for syndrome of retention of phlegm and blood stasis in the lung were 4.54, 12.70%, 55.17%; those for syndrome of qi deficiency of the lung and spleen were 3.25, 27.30%, 13.79%; those for syndrome of qi deficiency of the lung and kidney were 3.32, 27.24%, 13.79%; those for syndrome of qi-yin deficiency of the lung and kidney were 3.29, 28.98%, 24.14%; those for syndrome of yin deficiency of the lung and kidney were 2.98, 32.71%, 3.45%; those for syndrome of blood stasis were 4.67, 10.29%, 62.07% and those for syndrome of fu shi were 3.07, 30.01%, 6.90%, all respectively. The blood stasis and fu shi were accompanying syndromes. CONCLUSION: Seven common syndromes of AECOPD are wind and cold invasion of lung, exogenous cold-evil and fluid-retention, phlegm-heat obstruction in the lung, pulmonary stagnation of phlegm qi deficiency of the lung and spleen, qi deficiency of the lung and kidney, as well as qi-yin deficiency of the lung and kidney. One additional syndrome that accompanies many of the afore-mentioned syndromes was blood stasis.

Shenmai injection for chronic pulmonary heart disease: a systematic review and meta-analysis.

OBJECTIVES: This study sought to evaluate the efficacy and safety of shenmai injection for chronic pulmonary heart disease (CPHD). METHODS: A systematic review was conducted of clinical trials that compared shenmai injection plus conventional medicine treatment versus conventional medicine treatment alone. Randomized controlled trials of clinical therapeutic studies on CPHD by shenmai injection were included. Searches were applied to the following electronic databases: the PubMed (1977-2008), the Cochrane Library, EMBASE, AMED, Chinese BioMedical Literature Database, and CBM. No blinding and language restriction was used. Data were extracted independently by 2 reviewers. All trials included were analyzed according to the criteria of the Cochrane Handbook. Review Manager 5.0 software was used for data analysis. RESULTS: Thirty-three (33) randomized clinical trials (2617 patients) with low methodological quality were included. Compared to conventional medicine treatment alone, shenmai plus conventional medicine treatment showed significant improvement in New York Heart Association classification of clinical status (odds ratio 0.24; 95% confidence interval 0.19-0.30), five studies had reported adverse events. No serious adverse effects were reported in any of the included trials. CONCLUSIONS: While there is some evidence that suggests potential effectiveness of shenmai plus conventional medical treatment for CPHD, the results of this study were limited by the methodological flaws, unknowns in concealment of allocation, number of dropouts, and blinding methods in the studies. Long-term and high-quality studies are needed to provide clear evidence for the future use of shenmai injection.