Objective To investigate the expression of YWHAQ protein in gastric adenocarcinoma tissues and its correlation with clinical pathological features and prognosis. Methods A total of 127 patients with gastric cancer who underwent radical surgery were enrolled. Clinical data and postoperative cancer tissue samples were collected from the patients. Immunohistochemistry was used to detect the protein expression of YWHAQ in gastric adenocarcinoma tissues. The relationship between YWHAQ expression and clinical pathological features and prognosis was analyzed. Bioinformatics prediction was performed to identify potential pathways regulated by YWHAQ in gastric adenocarcinoma. A protein-protein interaction network for YWHAQ was constructed using the STRING database. Results YWHAQ gene expression was significantly higher in gastric adenocarcinoma tissues than in normal tissues (P<0.05). The expression level of the YWHAQ protein was significantly correlated with age, tumor invasion depth, lymph node metastasis, and tumor stage (P<0.05). Kaplan-Meier survival analysis showed that patients with high YWHAQ expression had significantly poorer long-term survival than those with low expression (P<0.0001). Cox regression analysis revealed that high YWHAQ expression and distant metastasis were independent risk factors for gastric cancer (HR>1, P<0.05). Enrichment analysis indicated that YWHAQ positively regulated processes such as cell cycle, angiogenesis, and DNA damage repair. Conclusion YWHAQ is highly expressed in gastric adenocarcinoma tissues and may be associated with the occurrence, invasion, and metastasis of gastric adenocarcinoma. High YWHAQ expression and distant metastasis are independent risk factors for the prognosis of gastric adenocarcinoma patients.

Objective To analyze the effect of LINC00894 on the malignant biological behavior of liver cancer cells by regulating the microRNA(miR)-495-3p/RAS associated binding protein 23(Rab23)axis.Methods Cancer and adjacent tissues of 53 patients with liver cancer who undergoing surgery from January 2023 to November 2023 were collected.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of LINC00894,miR-495-3p and Rab23 in tissues and in vitro cultured cells Huh7,SMMC-7721,HepG2 and L-02.Cells Huh7 were randomly separated into Huh7 group,NC group,si-LINC00894 group,anti-NC group,and anti-miR-495-3p group.CCK-8,flow cytometry and Transwell were used to compare the cell proliferation,apoptosis,migration and invasion of each group.Western blot was used to detect the expression of Rab23 and proliferation-related proteins in cells.The relationship between LINC00894 and miR-495-3p,miR-495-3p and Rab23 was analyzed and verified by dual luciferase reporter gene assay.Results The mRNA and protein expression of LINC00894 and Rab23 were higher in cancer tissues and cells of liver cancer patients,while the mRNA expression of miR-495-3p was lower(P＜0.05).The apoptosis rate of si-LINC00894 group was higher than that of Huh7 group and NC group,and the absorbance at 24 h,48 h,and 72 h,numbers of migrating and invading cells,and expression of Rab23 in cells were lower than those in Huh7 group and NC group(P＜0.05);the absorbance at 24 h,48 h,and 72 h,numbers of migrating and invading cells,and expression of Rab23 in cells in the anti-miR-495-3p group were higher than those in the anti-NC group and si-LINC00894 group,the apoptosis rate of cells was lower than that in the anti NC group and si-LINC00894 group(P＜0.05).The luciferase activity in the WT-LINC00894+miR-495-3p group was lower than that in the WT-LINC00894+miR-NC group(P＜0.05),the luciferase activity in the WT-Rab23+miR-495-3p group was lower than that in the WT-Rab23+miR-NC group(P＜0.05).Conclusion Downregulation of LINC00894 can promote the expression of miR-495-3p,inhibit the expression of Rab23,and thus exert the inhibitory effect on the malignant biological behavior of liver cancer cells.

OBJECTIVE To establish a visualized loop-mediated isothermal amplification(LAMP)for rapid detec-tion of Klebsiella pneumoniae and evaluate the sensitivity and specificity of the detection method.METHODS To-tally4 specific LAMP primers were designed by targeting to rcsA gene of the K.pneumoniae.The reaction condi-tions(temperature,time)and the parameters of the reaction system were optimized,the reaction was made visu-alized by using chimeric fluorescent dye SYBR Green Ⅰ.The optimal reactions conditions and reaction systems were determined based on the results of visualization and agarose gel electrophoresis.The optimized conditions and systems were used to test the sensitivity by diluting DNA template on a 10-fold gradient,meanwhile,the specifici-ty of the method was evaluated by detecting K.pneumoniae and other ten species of common bacteria and fungi.RESULTS The optimal reaction temperature was 63 ℃ after the optimization,with the reaction time 35 min;the concentration of buffer solution in the reaction system was determined as 0.8×,with the concentration of magne-sium ion 8 mmol/L,the concentration of dNTPs 1.4 mmol/L,the ratio of internal to external primers 6∶1,the concentration of Bst DNA polymerase 0.32 U/μl,the concentration of betaine 0.75 mmol/L.For the test of sensi-tivity,the method could detect the template with the concentration of 1.5 ng/μl.For the test of specificity,only the detection of K.pneumoniae could display positive visualized result and positive electrophoretic band.CONCLUSIONS The visualized LAMP with high specificity and sensitivity for rapid detection of K.pneumoniae is successfully established,facilitating the observation of the detection result without the use of precise instruments.It is suitable for the detection in grass-root laboratories after successful preliminary application in clinical detection.

Objective To analyze the effect of LINC00894 on the malignant biological behavior of liver cancer cells by regulating the microRNA(miR)-495-3p/RAS associated binding protein 23(Rab23)axis.Methods Cancer and adjacent tissues of 53 patients with liver cancer who undergoing surgery from January 2023 to November 2023 were collected.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of LINC00894,miR-495-3p and Rab23 in tissues and in vitro cultured cells Huh7,SMMC-7721,HepG2 and L-02.Cells Huh7 were randomly separated into Huh7 group,NC group,si-LINC00894 group,anti-NC group,and anti-miR-495-3p group.CCK-8,flow cytometry and Transwell were used to compare the cell proliferation,apoptosis,migration and invasion of each group.Western blot was used to detect the expression of Rab23 and proliferation-related proteins in cells.The relationship between LINC00894 and miR-495-3p,miR-495-3p and Rab23 was analyzed and verified by dual luciferase reporter gene assay.Results The mRNA and protein expression of LINC00894 and Rab23 were higher in cancer tissues and cells of liver cancer patients,while the mRNA expression of miR-495-3p was lower(P＜0.05).The apoptosis rate of si-LINC00894 group was higher than that of Huh7 group and NC group,and the absorbance at 24 h,48 h,and 72 h,numbers of migrating and invading cells,and expression of Rab23 in cells were lower than those in Huh7 group and NC group(P＜0.05);the absorbance at 24 h,48 h,and 72 h,numbers of migrating and invading cells,and expression of Rab23 in cells in the anti-miR-495-3p group were higher than those in the anti-NC group and si-LINC00894 group,the apoptosis rate of cells was lower than that in the anti NC group and si-LINC00894 group(P＜0.05).The luciferase activity in the WT-LINC00894+miR-495-3p group was lower than that in the WT-LINC00894+miR-NC group(P＜0.05),the luciferase activity in the WT-Rab23+miR-495-3p group was lower than that in the WT-Rab23+miR-NC group(P＜0.05).Conclusion Downregulation of LINC00894 can promote the expression of miR-495-3p,inhibit the expression of Rab23,and thus exert the inhibitory effect on the malignant biological behavior of liver cancer cells.

OBJECTIVE To establish a visualized loop-mediated isothermal amplification(LAMP)for rapid detec-tion of Klebsiella pneumoniae and evaluate the sensitivity and specificity of the detection method.METHODS To-tally4 specific LAMP primers were designed by targeting to rcsA gene of the K.pneumoniae.The reaction condi-tions(temperature,time)and the parameters of the reaction system were optimized,the reaction was made visu-alized by using chimeric fluorescent dye SYBR Green Ⅰ.The optimal reactions conditions and reaction systems were determined based on the results of visualization and agarose gel electrophoresis.The optimized conditions and systems were used to test the sensitivity by diluting DNA template on a 10-fold gradient,meanwhile,the specifici-ty of the method was evaluated by detecting K.pneumoniae and other ten species of common bacteria and fungi.RESULTS The optimal reaction temperature was 63 ℃ after the optimization,with the reaction time 35 min;the concentration of buffer solution in the reaction system was determined as 0.8×,with the concentration of magne-sium ion 8 mmol/L,the concentration of dNTPs 1.4 mmol/L,the ratio of internal to external primers 6∶1,the concentration of Bst DNA polymerase 0.32 U/μl,the concentration of betaine 0.75 mmol/L.For the test of sensi-tivity,the method could detect the template with the concentration of 1.5 ng/μl.For the test of specificity,only the detection of K.pneumoniae could display positive visualized result and positive electrophoretic band.CONCLUSIONS The visualized LAMP with high specificity and sensitivity for rapid detection of K.pneumoniae is successfully established,facilitating the observation of the detection result without the use of precise instruments.It is suitable for the detection in grass-root laboratories after successful preliminary application in clinical detection.

Objective:To explore the clinical application and triage management value of using blood circulating cell-free DNA (cfDNA) (cysteine dioxygenase type 1 gene, CDO1, and Homeobox protein A9 gene, HOXA9) hypermethylation level to detect and diagnose ovarian cancer.Methods:A case-control study was conducted on patients who went for surgery at Chengdu Womens and Childrens Central Hospital from November 2022 to October 2023. Blood samples were collected before surgery for evaluation of cancer antigen 125 (CA125), human epididymis protein 4 (HE4), risk of ovarian malignancy algorithm (ROMA) score, and DNA methylation testing. The basic clinical information, biomarkers, and transvaginal ultrasound (TVS) information were collected simultaneously. Information from a total of 151 patients was collected, including 122 cases with benign pathology and 29 ovarian cancer cases. The pathologic diagnosis of ovarian tissue was defined as the gold standard. The multivariate logistic regression analysis was used to identify high-risk factors for ovarian cancer. The clinical efficacy of DNA methylation detection for ovarian cancer was analyzed using the area under curve (AUC).Results:The results showed that the age, menopausal status, CA125 and HE4 detection, ROMA score, positivity rate of CDO1 gene and HOXA9 gene single or combined testing in ovarian cancer patients were higher than those in the benign group and showed significant differences ( P<0.05). Among these detection protocols, the AUC of CDO1 and HOXA9 dual gene methylation testing for ovarian cancer was the highest at 0.936 (95% CI, 0.878-0.994), with 89.7% (95% CI 73.6%-96.4%) sensitivity and 97.5% (95% CI 93.0%-99.2%) specificity, respectively. The positive detection rate of CDO1 and HOXA9 dual gene methylation in early ovarian cancer FOGO I-II stage is 12/14 higher than other tests. Conclusion:Blood cfDNA methylation detection, a simple, non-invasive, and highly sensitive detection method, is superior to the current ovarian cancer testing in the risk assessment and early detection.

Objective:To investigate the effects of thinned anterolateral thigh perforator flaps combined with finger splitting and webplasty in sequential treatment of degloving destructive wound of total hand.Methods:This study was a retrospective observational study. From January 2012 to January 2023, a total of 15 cases who met the inclusion criteria with degloving destructive wound of total hand were admitted to Ningbo No.6 Hospital, including 10 males and 5 females, aged 17-75 years. The wounds were all combined with exposed bones or tendon. Emergency debridement and vacuum sealing drainage were performed in all cases before flap transplantation in stage Ⅰ. After thorough debridement, the wound area was 11.0 cm×3.0 cm-23.0 cm×13.5 cm. One or both anterolateral thigh perforator flaps with size of 12.5 cm×5.0 cm-25.0 cm×15.5 cm were designed, cut, and thinned to repair the skin and soft tissue defects of the hand. The donor site was sutured directly or repaired with medium-thickness skin graft from the opposite thigh. As needed, the flap was reconstructed by finger splitting and webplasty once or more times every 3 months after stage Ⅰoperation. The survival and complications of flap and wound healing at the donor site were observed after stage Ⅰoperation. The appearance of flap, two-point discrimination distance, and hand function were observed during the follow-up. At the final follow-up, the function of the affected hand was evaluated by the trial standards for evaluation of partial function of upper extremity by the Hand Surgery Society of Chinese Medical Association.Results:After the operation of stage Ⅰ, all the flaps of 15 cases of patients survived completely, including 1 case that had arterial crisis of flap but survived completely after exploration and re-anastomosis of blood vessels; all the wounds at the donor site healed. During the follow-up period of 6 to 18 months after stage Ⅰ, the flap was slightly swollen, with a little pigmentation, and the two-point discrimination distance in the finger flap was 8-11 mm. The fingers could complete the basic life actions such as flexion, extension, pinch, and grip. At the final follow-up, 3 cases were excellent, 9 cases were good, and 3 cases were acceptable in function evaluation of the affected hand.Conclusions:For degloving destructive wound of total hand, free transplantation of one or both thinned anterolateral thigh perforator flaps is used for repair in stage Ⅰ, and finger splitting and webplasty are used to reconstruct the flaps in the later stage, which can basically restore the pinch and grip function of the affected hand that is required for daily life, and is worthy of clinical promotion.

Background and purpose:Human epidermal growth factor receptor 2(HER2)serves as one of the paramount drivers of breast cancer metastasis,with roughly 20%-30%of breast cancer patients exhibiting high expression of HER2.The expression level of HER2 is regulatable at multiple molecular levels and determines the metastatic potential of breast cancer cells;however,the manner in which HER2 expression is modulated at the mRNA level remains ambiguous.Circ-0007766 is a circRNA originated from the coding gene ERBB2 for HER2,and whether circ-0007766 can regulate HER2 expression via the ceRNA mechanism has not been reported.This study aimed to analyze whether circ-0007766 acts as a miR-1972 sponge to promote breast cancer cell migration and invasion via upregulation of HER2 expression.Methods:In this study,a high-throughput circRNA chip was employed to screen for circRNAs that exhibited highly specific expression in HER2-positive breast cancer cells.RNA fluorescence in situ hybridization(FISH)was utilized to detect the subcellular localization of circ-0007766.The BaseScope experiment was conducted to analyze the expression level of circ-0007766 in breast cancer tissues and its clinical diagnostic significance.Breast cancer cell models with overexpression and knockdown of circ-0007766 were constructed by transfecting cloning plasmids and siRNA in vitro.The effect of circ-0007766 on the migration and invasion of breast cancer cells was assessed using transwell migration and invasion experiments,and the migration and invasion abilities of MDA-MB-231 and SK-BR-3 cells were measured.Additionally,it was evaluated whether circ-0007766 could promote the migration and invasion of breast cancer cells through miR-1972.A dual luciferase reporter gene assay was used to verify whether circ-0007766 could regulate HER2 expression by binding to miR-1972.The direct interaction between circ-0007766 and miR-1972 was further verified through the RAP experiment.RIP detection was performed in MDA-MB-231 cells,and the relative 3'UTR of HER2 mRNA was measured by real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR).Western blot was used to detect the protein expressions.Results:Circ-0007766 was conspicuously highly expressed in HER2-positive breast cancer cells and distributed in both the cytoplasm and nucleus of cells,with the preponderance being in the cytoplasm.The expression level of circ-0007766 was strikingly higher in breast cancer tissues than in para-cancerous tissues.The expression of circ-0007766 was significantly elevated in HER2-positive breast cancer samples compared with HER2-negative samples.The overexpression(knockdown)of circ-0007766 in HER2-negative breast cancer cells(in HER2-positive breast cancer cells)was capable of promoting(inhibiting)the migration and invasion of breast cancer cells.Circ-0007766 directly bound to miR-1972,which inhibited breast cancer cell migration and invasion,thereby forming an endogenous competitive RNA(ceRNA)regulatory network and impeding the downregulation of HER2 mRNA and protein expression mediated by miR-1972.Circ-0007766 could potentiate the inhibitory effect of miR-1972 on HER2-mediated breast cancer cell migration and invasion that was negatively regulated by miR-1972.CircRNAs sequestered miRNAs to function as ceRNAs,thereby regulating gene expression at both the transcriptional and translational levels.Finally,we discovered that the expressions of circ-0007766 and HER2 were positively correlated in breast cancer cell and tissue samples,while the expression levels of miR-1972 and HER2 were negatively correlated.Circ-0007766 could specifically target miR-1972 to hinder its regulatory effect on HER2 expression.Conclusion:This study discovers that circ-0007766 facilitates the migration and invasion of breast cancer cells via the miR-1972/HER2 signal axis,offering a novel biomarker and potential therapeutic target for patients with metastatic HER2-positive breast cancer.

Objective To investigate the efficacy and safety of retroperitoneal single-port laparoscopic (LESS) adrenal tumor resection.Methods The clinical data of 130 patients receiving multi-port and single-port laparoscopic adrenal tumor resection in this hospital from January 2015 to March 2023 were retrospec-tively analyzed.There were 50 cases in the LESS group and 80 cases in the traditional laparoscopic adrenal tumor resection (LA) group.The age,gender,tumor location,complicating underlying diseases and tumor size were collected in the two groups.The related perioperative indicators were compared between the two groups. Results The operation time[(95±52)min vs. (101±58)min],drainage tube indwelling time[(4.9±1.5)d vs. (6.7±1.0)d],postoperative incision satisfaction degree and pain situation had statistically significant differences between the LESS group and LA group (P<0.05).The intraoperative bleeding volume[(20.2±13.2)mL vs. (25.6±11.3)mL]and hospitalization duration[(5.9±1.3)d vs. (7.8±1.0)d]had no statisti-cal differences between the two groups (P>0.05).No intraoperative and postoperative complications oc-curred in the two groups.There was 1 case of conversion to open operation in the LA group.Conclusion The LESS adrenal tumor resection is safe and effective,and the incision is more beautiful compared with multi-port laparoscopic operation.

A patient who suffered with degloving and destructive servered distal segment of the left thumb was referred to the Department of Hand Surgery, Ningbo No.6 Hospital in May 2021. The thumb was reconstructed by transfer and splicing of a free bilateral fibular hallux nail flap. Metatarsal artery of the metatarsal fibular side of the flap was anastomosed to the proper palmar digital artery at the recipient site. The nerve meridian were anastomosed to the proper palmar digital nerve of thumb, and the distal arterial arch of the metatarsal base between the flaps was anastomosed at the same time. A V-Y advancement flap was employed to repair the donor site. After 3 months, the transferred flaps survived well, the left thumb nail grew well, and the dynamic TPD was at 7 mm. According to the Evaluation Standard of Thumb and Finger Reconstruction Function of the Society of Hand Surgery of the Chinese Medical Association, it was evaluated as excellent (14 points). The advancement flap in the donor site survived well, and the walking, running and jumping of both feet were not affected. According to Maryland's foot function scoring standard, it was evaluated as excellent (98 points).