AIM:To investigate the regulatory effects of proanthocyanidins on autophagy and apoptosis in the retinas of guinea pigs with form-deprivation myopia via the AMPK/Wnt/β-catenin pathway.METHODS:Fifty guinea pigs were randomly divided into a normal control group, a myopia model group, and low-dose, medium-dose, and high-dose proanthocyanidins groups(25, 50 and 100 mg/kg). Refractive power and axial length of right eye were measured using a retinoscope and A-scan ultrasound. Retinal pathological changes were observed via HE staining. Immunohistochemistry assessed p-AMPK and p-mTOR expression in the retina. Immunofluorescence detected p62 and LC3 expression. TUNEL staining evaluated retinal cell apoptosis. Western blot examined expression of proteins related to the AMPK/Wnt/β-catenin pathway and autophagy(p62, Beclin1, LC3-II/LC3-I), and apoptosis-related proteins(Bax, Bcl-2, Cleaved-Caspase3, Caspase3)in the retina.RESULTS:Compared with the control group, the myopia model group showed significantly reduced refractive power and significantly increased axial length(both P<0.05); retinal cell arrangement became sparse and retinal thickness thinned. The p-AMPK levels in the retina were significantly reduced, while p-mTOR levels were significantly increased(both P<0.05), indicating suppression of the AMPK-Wnt/β-catenin pathway. The p62 levels were significantly elevated and LC3 levels were significantly reduced(both P<0.05), suggesting inhibition of autophagy. Bax and Cleaved-Caspase3 were significantly increased, while Bcl-2 was significantly decreased, indicating significantly increased apoptosis(both P<0.05). Compared with the myopia model group, all proanthocyanidins dose groups significantly inhibited refractive error reduction and axial length growth(both P<0.05), restored retinal cell alignment and thickness, activated the AMPK/Wnt/β-catenin pathway, significantly increased p-AMPK expression, and suppressed p-mTOR expression(all P<0.05); significantly suppressed p62 expression, increased Beclin1 and LC3-II/LC3-I expression(both P<0.05), and activated retinal autophagy; significantly suppressed Bax and Cleaved-Caspase3 expression, increased Bcl-2 expression(both P<0.05), and inhibited retinal cell apoptosis.CONCLUSION:Proanthocyanidins enhance retinal autophagy by activating the AMPK/Wnt/β-catenin pathway, thereby inhibiting retinal apoptosis and preventing or alleviating the onset of myopia.

Objective: To understand the needs of parents of kindergarten children in urban areas for online training on sexuality education, so as to provide a basis for the development of parent training courses. Methods: In May 2023, a multistage cluster random sampling method was used to select 3 516 parents of young children from 12 kindergartens in urban areas of Changshou District, Chongqing. A self designed questionnaire on Knowledge, Attitude and Practice of Early Childhood Sexuality Education (Parents) was used for the survey, and χ 2 test and multi factor Logistic regression model were used for data analysis. Results: The online training demand rate of urban parents for early childhood sexuality education was 57.05%. The results of the multi factor Logistic regression analysis showed that age, education level, occupation, whether or not they were the main caregivers, total score level of the scale, awareness of their own responsibility, communication with family and friends about early childhood sexuality education and young children s participation in kindergarten sexuality education activities were the influencing factors of parents online training demand on sexuality education ( OR =1.18, 1.44, 1.42, 0.83, 1.19, 0.51, 0.75, 0.75, P <0.05). Conclusion Urban parents have a high demand for online training on early childhood sexuality education, and training courses should be developed according to the specific needs and characteristics of parents of young children.

【Objective】 To investigate the effect of total flavone of oldenlandia diffusa(FOD) on the stemness, proliferation and apoptosis of breast cancer(BC) stem cells sorted from MDA-MB-231. 【Methods】 Human BC cell lines MDA-MB-231 was cultured in vitro; MDA-MB-231 was stimulated by different concentrations(0 μg/mL, 100 μg/mL, 200 μg/mL and 400 μg/mL) of FOD for different time (24 h, 48 h and 72 h). CCK8 and plate cell cloning assay were used to detect the effect of FOD on MDA-MB-231 proliferation; CD44⁺/CD24^-MDA-MB-231 cell line were tested by flow cytometry and stem cell markers such as Nanog, Oct4 and Sox2 were tested by Western blotting; Annexin V-PE/7-AAD was used to detect the effect of FOD on MDA-MB-231 apoptosis and Bcl2, cleaved-caspase3 and Bax were tested by Western blotting. 【Results】 Cell proliferation of MDA-MB-231 was significantly inhibited by FOD, with the significant suppression at concentrations of 400 μg/mL for 72 h compared with negative control group(P<0.05). The apoptosis rate was significantly upregulated than the negative control group (P<0.05). The protein expression of Bcl2 decreased while Bax and cleaved-caspae3 increased, and stemness markers such as Nanog, Sox2 and Oct4 decreased in FOD-treated cells. Moverover, Akt-GSK3β-β-catenin axis was inhibited in FOD-treated cells. 【Conclusion】 FOD could significantly inhibit the stemness and proliferation and promote the apoptosis of MDA-MB-231.

【Objective】 To investigate the effects of total flavone of oldenlandia diffusa (FOD) on the proliferation and apoptosis of hepatocellular carcinoma (HCC) stem cells sorted from Huh7. 【Methods】 Human HCC cell lines Huh7 was cultured in vitro; CD133 positive (CD133⁺) stem cells in Huh7 cell line were sorted by flow cytometry, and stem cell markers such as Nanog, Oct4 and Sox2 were tested by Western blotting. CD133⁺-Huh7 was stimulated by different concentrations (0 μg/mL, 50 μg/mL, 100 μg/mL and 400 μg/mL) of FOD for different time (24 h, 48 h, 72 h and 96 h). CCK8 and plate cell cloning assay were used to detect the effect of FOD on CD133⁺-Huh7 proliferation while Annexin V-PE/7-AAD was used to detect the effect of FOD on CD133⁺-Huh7 apoptosis. Western blotting was used to detect the protein expressions of protein 53 (P53), factor associated suicide-Fas-associating protein with a novel death domain (Fas-FADD), B-cell lymphoma-2 (Bcl-2), Cleaved-Caspase3, and Bcl-2 associated X protein (Bax). 【Results】 More than 95% of stem cells were purified for further experiments. Cell proliferation of CD133⁺-Huh7 was significantly inhibited by FOD, with the significant suppression at the concentration of 100 μg/mL for 72 h compared with negative control group (P<0.05). The apoptosis rate was significantly upregulated than that in the negative control group (P<0.05). The protein expression of Bcl2 decreased while Bax and Cleaved-Caspae3 increased via FAS/FADDD and P53 axis. 【Conclusion】 FOD can significantly inhibit the proliferation and promote the apoptosis of CD133⁺-Huh7.

Tumor metastasis depends on the dynamic balance of the actomyosin cytoskeleton. As a key component of actomyosin filaments, non-muscle myosin-IIA disassembly contributes to tumor cell spreading and migration. However, its regulatory mechanism in tumor migration and invasion is poorly understood. Here, we found that oncoprotein hepatitis B X-interacting protein (HBXIP) blocked the myosin-IIA assemble state promoting breast cancer cell migration. Mechanistically, mass spectrometry analysis, co-immunoprecipitation assay and GST-pull down assay proved that HBXIP directly interacted with the assembly-competent domain (ACD) of non-muscle heavy chain myosin-IIA (NMHC-IIA). The interaction was enhanced by NMHC-IIA S1916 phosphorylation via HBXIP-recruited protein kinase PKCβII. Moreover, HBXIP induced the transcription of PRKCB, encoding PKCβII, by coactivating Sp1, and triggered PKCβII kinase activity. Interestingly, RNA sequencing and mouse metastasis model indicated that the anti-hyperlipidemic drug bezafibrate (BZF) suppressed breast cancer metastasis via inhibiting PKCβII-mediated NMHC-IIA phosphorylation in vitro and in vivo. We reveal a novel mechanism by which HBXIP promotes myosin-IIA disassembly via interacting and phosphorylating NMHC-IIA, and BZF can serve as an effective anti-metastatic drug in breast cancer.

Objective: On the basis of the dominant frequency index of functional connectivity, the "brain age" analysis method was used to explore abnormal development patterns of sensorimotor networks in boys with autism spectrum disorder(ASD). Methods: The resting state functional magnetic resonance data (7-12 years old) for 105 boys with ASD and 102 matched boys with normal development from the ABIDE public database were screened. Functional connection networks in different frequency bands of sensorimotor related brain regions were constructed for each individual, and the frequency of the strongest connection were constructed as the optimal frequency of the connection. Brain age analysis was used to explore the difference between brain age and chronological age in boys with ASD. Results: The brain sensorimotor network of boys with ASD showed an abnormal development pattern of overdevelopment followed by underdevelopment, and the transition between the two patterns occurred at approximately 7.8 years of age. Older boys with ASD (older than 10 years) whose underdevelopment trend was suppressed had lower ASD severity( r=-0.43, P < 0.05 ). Conclusion The brain sensorimotor network in boys with ASD has an abnormal development process, and the brain chronological age difference in the sensorimotor network has potential as a neuroimaging marker to measure the development of ASD.

ObjectiveTo observe the effect of Simiaowan on the intestinal ATP-binding cassette superfamily G （White） member 2 （ABCG2） expression and the intestinal uric acid excretion in hyperuricemia rats. MethodA total of 48 SD male rats were randomized into the normal， model， benzbromarone （4.7 mg·kg^-1）， and high-， medium-， low-dose Simiaowan groups （2 260.6， 1 130.3， 565.2 mg·kg^-1， respectively）， with 8 rats in each group. Potassium oxonate and hypoxanthine was employed to induce hyperuricemia in rats （21 days）. On the 8^th day， administration began （once a day for 14 days）. Rats were killed on the 21^st day， and serum uric acid， serum creatinine， blood urea nitrogen， and intestinal uric acid were detected. The protein expression of ABCG2 in the small intestine was detected by Western blot. The ABCG2 protein expression and localization in intestinal tissues were determined by immunohistochemistry. The ABCG2 mRNA expression in small intestine was measured by quantitative real-time PCR. ResultThe levels of serum uric acid， serum creatinine， and blood urea nitrogen in the model group were higher than those in the normal group （P<0.01）. Low level of serum uric acid in the three Simiaowan groups and benzbromarone group （P<0.01）， high level of intestinal uric acid in medium-dose and low-dose Simiaowan groups （P<0.05， P<0.01）， high level of serum creatinine in benzbromarone group （P<0.01）， and low level of blood urea nitrogen in low-dose Simiaowan group （P<0.05） were observed as compared with those in the model group. Serum uric acid showed insignificant difference between the low-dose Simiaowan group and benzbromarone group. The expression of ABCG2 protein in the model group was lower than that in the normal group （P<0.05）. The expression of ABCG2 protein in the medium-dose and low-dose Simiaowan groups （P<0.05， P<0.01）， the high-dose Simiaowan group， and benzbromarone group increased as compared with that in the model group. ABCG2 mRNA expression was insignificantly different between the model group and the normal group， while the expression in the medium-dose and low-dose Simiaowan groups was higher than that in the model group （P<0.05）. ABCG2 protein was mainly distributed in intestinal villi， and ABCG2 protein expression demonstrated no significant difference between the model group and the normal group. The ABCG2 protein expression in the three Simiaowan groups increased as compared with that in the model group （P<0.05）. ConclusionSimiaowan can significantly reduce the serum uric acid level in hyperuricemia rats. Particularly， the low-dose Simiaowan shows similar efficacy to benzbromarone in lowering uric acid and protects renal function. The mechanism is the likelihood that it up-regulates intestinal ABCG2 expression to promote intestinal excretion of uric acid.

Conversion therapy has become the core in the treatment of borderline resectable or unresectable liver cancer, which provides resectable opportunities for more advanced liver cancer patients. In accordance with the first-choice treatment regimen recommended by the guidelines, the authors reported a successful case of Atezolizumab and Bevacizumab (T+A regimen) conversion therapy. The patient with initially borderline resectable advanced liver cancer was performed liver segment resection sucessfully after conversion therapy, and non-tumor recurrence was observed at postoperative 9 months. Postoperative pathological examination showed combined hepatocellular-cholangiocarcinoma, which also indicated the important value of T+A regimen in the conversion therapy of combined hepatocellular-cholangiocarcinoma.

Objective:To compare the therapeutic effect of lateral position and half lithotomy position in Asian proximal femur intramedullary nail antirotation system (PFNA-II) for treating the elderly patients with femoral intertrochanteric fractures.Methods:A retrospective case control study was made on 141 patients with femoral intertrochanteric fractures admitted to Jiangjin Central Hospital from January 2016 to September 2017, including 54 males and 87 females, aged 65-99 years (mean, 80.4 years). According to AO classification, there were 42 patients with type A1 fractures, 88 with type A2 and 11 with type A3. Of all, 74 patients were stabilized by PFNA-II internal fixation in lateral position (lateral position group) and 67 patients by PFNA-II internal fixation in half lithotomy position (half lithotomy position group). The postural placement time, total incision length, operative time, intraoperative blood loss, fluoroscopy frequency, tip-apex distance, reduction quality, fracture healing time, postoperative complications and Harris hip function at 12 months after surgery were compared between the two groups.Results:All patients were followed up for 12-18 months (mean, 12.5 months), except that 13 patients were lost after 9 months, an average of 12.5 months. There were no statistically significant differences in postural placement time, operative time, fracture healing time, and Harris hip score between the two groups ( P>0.05). While significant differences were seen between lateral position group and half lithotomy position group regarding the incision length [(6.5±1.3)cm vs. (7.5±1.5)cm], intraoperative blood loss [(84.3±3.1)ml vs. (90.4±3.9)ml], fluoroscopy frequency [(13.1±1.9)times vs. (11.2±1.2)times], tip-apex distance [(20. 6±2.2)mm vs. (24.4±1.8)mm], good rate of reduction quality (80% vs. 85%) and implant related complications (5% vs. 2%) ( P<0.05 or 0.01). Conclusion:For treatment of elderly patients with intertrochanteric fractures, compared to the lateral position, the half lithotomy position in PFNA-II internal fixation can reduce frequency of fluoroscopy, improve quality of fracture reduction and reduce implant-related complications.

Microsatellite instability (MSI) is a key biomarker for cancer therapy and prognosis. Tra-ditional experimental assays are laborious and time-consuming, and next-generation sequencing-based computational methods do not work on leukemia samples, paraffin-embedded samples, or patient-derived xenografts/organoids, due to the requirement of matched normal samples. Herein, we developed MSIsensor-pro, an open-source single sample MSI scoring method for research and clinical applications. MSIsensor-pro introduces a multinomial distribution model to quantify poly-merase slippages for each tumor sample and a discriminative site selection method to enable MSI detection without matched normal samples. We demonstrate that MSIsensor-pro is an ultrafast, accurate, and robust MSI calling method. Using samples with various sequencing depths and tumor purities, MSIsensor-pro significantly outperformed the current leading methods in both accuracyand computational cost. MSIsensor-pro is available at https://github.com/xjtu-omics/msisensor-pro and free for non-commercial use, while a commercial license is provided upon request.