Identification of COQ2 as a regulator of proliferation and lipid peroxidation through genome-scale CRISPR-Cas9 screening in myeloma cells.

Multiple myeloma (MM) is the second most common malignant haematological disease with a poor prognosis. The limit therapeutic progress has been made in MM patients with cancer relapse, necessitating deeper research into the molecular mechanisms underlying its occurrence and development. A genome-wide CRISPR-Cas9 loss-of-function screening was utilized to identify potential therapeutic targets in our research. We revealed that COQ2 plays a crucial role in regulating MM cell proliferation and lipid peroxidation (LPO). Knockout of COQ2 inhibited cell proliferation, induced cell cycle arrest and reduced tumour growth in vivo. Mechanistically, COQ2 promoted the activation of the MEK/ERK cascade, which in turn stabilized and activated MYC protein. Moreover, we found that COQ2-deficient MM cells increased sensitivity to the LPO activator, RSL3. Using an inhibitor targeting COQ2 by 4-CBA enhanced the sensitivity to RSL3 in primary CD138+ myeloma cells and in a xenograft mouse model. Nevertheless, co-treatment of 4-CBA and RSL3 induced cell death in bortezomib-resistant MM cells. Together, our findings suggest that COQ2 promotes cell proliferation and tumour growth through the activation of the MEK/ERK/MYC axis and targeting COQ2 could enhance the sensitivity to ferroptosis in MM cells, which may be a promising therapeutic strategy for the treatment of MM patients.

[Research progress on Pheretima and earthworms with related origin].

Through literature analysis of Pheretima and its origin-related earthworm,this study summarized the progress on Pheretima in textual criticism of origin,origin identification,effective components,detection of harmful components,and pharmacological effects,which can lay a basis for further research on Pheretima. Through literature research,the authors found that Pheretima was first recorded in Secret Formulary for Traumatology and Fracture Taught by Immortal written by LIN Daoren in Tang Dynasty rather than the Taiping Holy Prescriptions for Universal Relief in Song Dynasty. The latest techniques for origin identification include microscopic trait identification,DNA barcoding,and HPLC. The main effective components of Pheretima are proteins,polypeptides,enzymes,nucleotides,amino acids,and trace elements. According to recent studies,Pheretima has anti-pulmonary and anti-renal interstitial fibrosis,respiratory syncytial virus-inhibiting,human hypertrophic scar fibroblast proliferation-suppressing,and mouse embryonic fibroblast proliferation-promoting effects. Moreover,Pheretima can prevent colitis-induced colon cancer by inhibiting the activation of COX-2/PGE2/ß-catenin signaling pathway. METHODS:: for detecting the harmful components and their residues( organic pollutant polychlorinated biphenyl,heavy metals) and bacteria in Pheretima,have been established. Pheretima,mainly derived from wild earthworms,has remarkable clinical efficacy. However,the wild resource is in short supply and artificial culture is expected to be a promising solution.

Amplification and overexpression of TP63 and MYC as biomarkers for transition of cervical intraepithelial neoplasia to cervical cancer.

OBJECTIVE: Biopsy confirmed that cervical intraepithelial neoplasia (CIN) may naturally regress or progress. Currently, the risk assessment for CIN progression to cervical cancer is still not satisfactory in clinical practice. We investigated copy number and protein expression of TP63 and MYC and explored the possibility to use them as progression biomarkers. METHODS: Copy numbers of TP63 and MYC, as well as human papilloma virus (HPV) integration status, were determined by fluorescence in situ hybridization in 39 patients with CIN and 66 patients with cervical cancer. Corresponding protein expressions were analyzed by immunohistochemistry. Receiver operating characteristic curves were used to measure the diagnostic test performance for the detection of cervical cancer from CIN. Sensitivity and specificity values of biomarkers were calculated. RESULTS: The average copy number and expression of TP63 and MYC, as well as the HPV integration rate, increased in the progression of CIN to cervical cancer. Receiver operating characteristic analysis for detection of cervical cancer resulted in area under the curve (AUC) values of TP63 copy number (AUC, 0.96; 95% confidence interval [CI], 0.91-1.00), MYC copy number (AUC, 0.92; 95% CI, 0.85-0.96), TP63 expression (AUC, 0.73; 95% CI, 0.61-0.85), and HPV-16 integration (AUC, 0.73; 95% CI, 0.60-0.85). MYC expression was not able to statistically distinguish cancer from CIN (P = 0.393). The combinations increased the specificity slightly but not sensitivity. Among them, TP63 amplification showed the best diagnostic performance. CONCLUSIONS: Amplification and overexpression of TP63 and MYC, and HPV integration rate, are associated with the transition of CIN to cervical cancer. Future studies on these biomarkers will help to assess the risk of CIN progression.

[Integration and demonstration of key techniques in surveillance and forecast of schistosomiasis in Jiangsu Province III Development of a machine simultaneously integrating mechanized environmental cleaning and automatic mollusciciding].

OBJECTIVE: To develop a machine simultaneously integrating mechanized environmental cleaning and automatic mollusciciding and to evaluate its effectiveness of field application, so as to provide a novel Oncomelania hupensis snail control technique in the large-scale marshlands. METHODS: The machine simultaneously integrating mechanized environmental cleaning and automatic mollusciciding, which was suitable for use in complex marshland areas, was developed according to the mechanization and automation principles, and was used for O. hupensis snail control in the marshland. The effect of the machine on environmental cleaning and plough was evaluated, and the distribution of living snails was observed at various soil layers following plough. The snail control effects of plough alone and plough followed by mollusciciding were compared. RESULTS: The machine could simultaneously complete the procedures of getting vegetation down and cut vegetation into pieces, plough and snail control by spraying niclosamide. After plough, the constituent ratios of living snails were 36.31%, 25.60%, 22.62% and 15.48% in the soil layers at depths of 0-5, 6-10, 11-15 cm and 16-20 cm respectively, and 61.91% living snails were found in the 0-10 cm soil layers. Seven and fifteen days after the experiment, the mortality rates of snails were 9.38% and 8.29% in the plough alone group, and 63.04% and 80.70% in the plough + mollusciciding group respectively (χ²7 d = 42.74, χ²15 d = 155.56, both P values < 0.01). Thirty days after the experiment, the densities of snails were 3.02 snails/0.1 m² and 0.53 snails/ 0.1 m² in the soil surface of the plough alone group and the plough + mollusciciding group, which decreased by 64.92% and 93.60%, respectively, and the decrease rate of snail density was approximately 30% higher in the plough + mollusciciding group than that in the plough alone group. CONCLUSIONS: The machine simultaneously integrating mechanized environmental cleaning and automatic mollusciciding achieves the integration of mechanical environmental cleaning and automatic niclosamide spraying in the complex marshland areas, which provides a novel technique of field snail control in the large-scale setting in China.

Determination of base binding strength and base stacking interaction of DNA duplex using atomic force microscope.

As one of the most crucial properties of DNA, the structural stability and the mechanical strength are attracting a great attention. Here, we take advantage of high force resolution and high special resolution of Atom Force Microscope and investigate the mechanical force of DNA duplexes. To evaluate the base pair hydrogen bond strength and base stacking force in DNA strands, we designed two modes (unzipping and stretching) for the measurement rupture forces. Employing k-means clustering algorithm, the ruptured force are clustered and the mean values are estimated. We assessed the influence of experimental parameters and performed the force evaluation for DNA duplexes of pure dG/dC and dA/dT base pairs. The base binding strength of single dG/dC and single dA/dT were estimated to be 20.0 ± 0.2 pN and 14.0 ± 0.3 pN, respectively, and the base stacking interaction was estimated to be 2.0 ± 0.1 pN. Our results provide valuable information about the quantitative evaluation of the mechanical properties of the DNA duplexes.

Left ventricular remodeling and fibrosis: Sex differences and relationship with diastolic function in hypertrophic cardiomyopathy.

OBJECTIVES: We investigated sex differences in left ventricular (LV) remodeling and fibrosis and their relationship with LV diastolic dysfunction by cardiovascular magnetic resonance (CMR). METHODS: CMR imaging was performed simultaneously in 152 age-matched patients (76 men, 76 women; mean age: 49±9 years) without LV systolic dysfunction. LV remodeling index (LVRI) was calculated as the ratio of LV mass and end-diastolic volume. Diastolic function indexes including peak filling rate (PFR) and time to PFR (tPFR) were evaluated. Extent of late gadolinium enhancement (LGE) was measured. RESULTS: LVRI and extent of LGE were greater in women compared with men (1.48±0.22 vs. 1.36±0.28g/ml; 13.15±2.48 vs. 11.35±2.34g, respectively, both P<0.001). Women had lower PFR and higher tPFR (both P<0.001) than men. LVRI and the extent of LGE showed significant relationships with parameters of diastolic function in both sex. In a multivariate analysis, LVRI remained a strong independent predictor of PFR and TPFR in women (ß=-0.272, P=0.032; ß=0.348, P=0.016, respectively), and in men (ß=-0.374, P<0.001; ß=0.660, P<0.001, respectively). Furthermore, the extent of LGE also remained an independent predictor of PFR in women (ß=-0.283, P=0.033) and men (ß=-0.492, P<0.001). CONCLUSIONS: There are prominent sex differences in LV remodeling and myocardial fibrosis. We suggest that the effects of LV remodeling and fibrosis may lead to diastolic dysfunction with greater susceptibility to worse clinical outcome in women.

Biventricular reverse remodeling after successful alcohol septal ablation for obstructive hypertrophic cardiomyopathy.

The aim of the present study was to investigate the long-term effects of alcohol septal ablation (ASA) on left ventricular (LV) and right ventricular (RV) remodeling in patients with obstructive hypertrophic cardiomyopathy (HC) using cardiovascular magnetic resonance (CMR). CMR was performed at baseline and 16 months after ASA in 38 patients with obstructive HC (mean age 48 ± 9 years) despite optimal medical treatment. ASA resulted in significant reductions of LV outflow tract gradient (mean 89 ± 22 vs 24 ± 12 mm Hg, p <0.001) and improvements in New York Heart Association functional class (p <0.001) during the follow-up period. LV remote mass and septal mass decreased from 98.34 ± 37.02 to 84.23 ± 34.71 g and from 77.56 ± 16.40 to 68.43 ± 14.02 g, respectively (p <0.001 for both) at 16-month follow-up. There were significant reductions of RV mass (mean 53.69 ± 7.12 vs 47.49 ± 6.17 g, p <0.001) and improvements in RV end-diastolic volume (mean 110.58 ± 22.47 vs 124.22 ± 24.17 ml, p <0.001) and the RV ejection fraction (p <0.001) during 16-month follow-up. Linear regression analysis showed that LV outflow tract gradient reduction was correlated significantly with LV remote mass reduction (r = 0.475, p = 0.003) and RV mass reduction (r = 0.535, p = 0.001) at 16-month follow-up. In conclusion, successful ASA can lead to positive biventricular reverse remodeling, showing significant reductions of RV and LV mass as well as increased RV and LV end-diastolic volumes during follow-up.

[Surveillance of schistosomiasis in Yanjiang Village, Hanjiang District, 2005-2010].

The endemic situation of schistosomiasis and Oncomelania snail status in Yanjiang Village, Hanjiang District decreased significantly from 2005 to 2010. However, the surveillance in key population such as fishmen and boatmen still needs to be strengthened.

[Approach to the road of vigorously developing Chinese medicine by scientific location of channels and collaterals].

Through researches of channels and collaterals and clinical practice of many years, the authors understand that Chinese medicine, which considers the human body as an interrelated, mutual constraints, whole, dynamic living system, has gradually become an important part of modern medicine. Channels and collaterals are a closed loop system which is communicated and linked by energy and information in the form of electromagnetic oscillation, reflecting many characteristics similar to quantum. Channels and collaterals are not a fixed organizational structure. Studies on channel and collaterals find that the track of the propagated sensation along channels (PSC) have the phenomenon drifting about. This exactly reflects the law of channels dynamically running. By information triggering and living resonance, channels and collaterals bring into play entirely regulative action. The innovative treatment of channels and collaterals followed by characteristics and laws of quantum can get a better curative effect. Theory of channels in the position of quantum information medical science provides an important breach for modernization of Chinese medicine.

[Effects of PML-RARalpha on cAMP-induced AML cell differentiation].

To explore the molecular mechanisms of acute promyelocytic leukemia cell differentiation induced by cAMP combined with low-dose As2O3, the PR9 cell line, which was stably transfected by PML-RARa fusion gene, was used as in vitro model. The effects of PML-RARa on cAMP-induced AML cell differentiation were evaluated according to cell growth, cell morphology, cell surface antigen as well as luciferase reporter gene assay, in the cells before and after the treatment with cAMP and/or As2O3. The results showed that cAMP alone could slightly increase the expression of CD11b in the PR9 cells expressing the PML-RARa fusion protein, but could not induce these cells to differentiate. The cells presented the terminal differentiation morphology and significantly increased CD11b expression only under the treatment of cAMP combined with As2O3. In addition, PML-RARa had strong inhibitory activity on the transcription of the reporter gene containing cAMP response elements. In conclusions, the PML-RARa fusion protein could dramatically block the signaling pathway of cAMP during the AML cell differentiation.