Objective To analyze survival outcomes and influencing factors among patients with colorectal cancer in Yunnan Province. Methods Clinical data were retrospectively collected from 4 892 patients with colorectal cancer. Survival data were obtained through follow-up. Overall survival (OS) was calculated by using the Kaplan-Meier method. Univariate analysis was performed by applying the log-rank test. Meanwhile, multivariate analysis employed the Cox proportional hazards regression model. Results The 1-, 3-, 5-, and 10-year OS rates for the entire cohort were 91.90%, 74.40%, 64.40%, and 28.70%, respectively. Univariate analysis revealed that age, ethnicity, region, differentiation grade, TNM stage, clinical stage, metastatic status, histological type, and treatment modality (chemotherapy, radiotherapy, and surgery) were associated with patient prognosis (all P<0.05). Multivariate analysis identified age (HR=1.250), region (HR=1.262), differentiation grade (HR=0.761), clinical stage (HR=3.128), and treatment modality (chemotherapy, HR=0.644; radiotherapy, HR=1.605; surgery, HR=0.384) as independent factors affecting survival prognosis in patients with colorectal cancer (all P<0.001). Conclusion Age, region, clinical stage, and treatment modality are independent factors influencing survival among patients with colorectal cancer in Yunnan Province. In clinical practice, these factors should be integrated to develop individualized prevention and treatment strategies, thereby improving patient outcomes.

To prepare antibodies that specifically recognize the conserved domain in the large extracellular loop of the CD63 protein, we expressed anti-CD63 single-chain variable fragment (scFv) antibody in Pichia pastoris in a secreted form. The purified expression product was found to bind specifically with CD63 protein and recognize CD63 on the surface of SK-MEL-28 cells. The variable region of the anti-CD63 monoclonal antibody in an anti-CD63-positive cell line was sequenced. The anti-CD63 scFv consisted of a variable heavy chain and a variable light chain linked by a flexible peptide was then designed. After codon optimization, the gene was synthesized and cloned into the expression plasmid pPICZα-A. The SacI-linearized plasmid was electroporated into P. pastoris X33, and 1% methanol were used to induce the expression of scFv. The fermentation supernatant was purified by Ni column. Anti-CD63 scFv was identified by SDS-PAGE and Western blotting, and its biological activities were analyzed by immunoblotting, immunofluorescence, cell-based ELISA, and flow cytometry. A P. pastoris strain capable of expressing and secreting anti-CD63 scFv was successfully obtained. The antibody had a molecular weight of approximately 30 kDa and specifically recognized CD63 protein. The expression of anti-CD63 scFv in P. pastoris paves the way for the production of anti-CD63 antibodies on a large-scale, which is undoubtedly an economical and effective way of antibody acquisition.
Single-Chain Antibodies/immunology* ; Humans ; Tetraspanin 30/immunology* ; Recombinant Proteins/immunology* ; Pichia/genetics* ; Saccharomycetales/metabolism*

Minimal residual disease (MRD), a crucial biomarker for assessing efficacy and predicting recurrence, refers to residual tumor cells remaining in the body of patients with hematological malignancies who achieved complete remission after treatment. This study aimed to conduct a retrospective analysis of the clinical diagnosis, treatment, and MRD monitoring of a pediatric patient with multiple acute B-lymphocytic leukemia relapses, alongside a review of relevant literature. In this case, Ig rearrangement based on next-generation sequencing (NGS) was more accurate in assessing the MRD level, compared with the traditional method of MRD detection, indicating the risk of earlier relapse and guided interventions in time. Additionally, NGS-MRD detected clonal evolution, providing new ideas to further investigate the intrinsic factors of disease development.

Minimal residual disease (MRD), a crucial biomarker for assessing efficacy and predicting recurrence, refers to residual tumor cells remaining in the body of patients with hematological malignancies who achieved complete remission after treatment. This study aimed to conduct a retrospective analysis of the clinical diagnosis, treatment, and MRD monitoring of a pediatric patient with multiple acute B-lymphocytic leukemia relapses, alongside a review of relevant literature. In this case, Ig rearrangement based on next-generation sequencing (NGS) was more accurate in assessing the MRD level, compared with the traditional method of MRD detection, indicating the risk of earlier relapse and guided interventions in time. Additionally, NGS-MRD detected clonal evolution, providing new ideas to further investigate the intrinsic factors of disease development.

Objective:To investigate the value of artificial intelligence (AI) cytomorphologic analysis system in the cytomorphological diagnosis and therapeutic evaluation of acute myeloid leukemia (AML).Methods:Bone marrow smear samples were collected from 150 patients with newly diagnosed and treated acute myeloid leukemia who were inpatients and outpatients at the Department of Institute of Hematology and Blood Diseases Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College from June 1, 2021 to July 31, 2022 for retrospective analysis. Among them, there were 50 patients in the newly diagnosed group, including 28 males and 22 females, with the onset age of 43.5(32.3,58.8)years. There were 100 patients in the post-treatment group, including 36 males and 64 females, with the onset age of 34.5(23.0,47.0)years. The results from cytomorphology expert were used as the gold standard and the Python 3.6.7 was used for analysis to evaluate the accuracy, sensitivity, and specificity of the AI cytomorphologic analysis system for blast cell recognition in AML diagnosis and treatment.Results:The proportion of blasts in AI analysis of 50 samples in the newly diagnosed group was≥20%, which met the diagnostic criteria of AML. AI analysis of blasts had an accuracy of 90.3%, sensitivity of 85.5%, and specificity of 98.0%. The correlation coefficient between AI and the proportion of blasts analyzed by experts was positively correlated( r=0.882, P<0.001). Meanwhile, in the post-treatment group, the sensitivity and specificity of AI analysis of blasts were 89.7% and 99.2%, respectively. The correlation coefficient between AI and the proportion of blasts analyzed by experts was positively correlated( r=0.957, P<0.001). According to AI analysis data, there are 8 samples in this group whose AI efficacy evaluation results on AML are inconsistent with expert analysis. Conclusion:AI cytomorphologic analysis system has high accuracy, sensitivity and specificity for blast cell recognition in AML morphological diagnosis and therapeutic evaluation.

Pancreaticoduodenectomy is one of the most difficult abdominal operations, and the difficulty in resection and complicated digestive tract reconstruction have brought great challenges for surgeons. At present, laparoscopic pancreaticoduodenectomy has been widely used in clinical practice, and compared with traditional 2D laparoscopy, 3D laparoscopy has the features of high magnification, high definition, and three-dimensional vision, which enables surgeons to see more clearly and operate more accurately, and thus it has great potential to be widely used in pancreaticoduodenectomy.

OBJECTIVE: To investigate the effect of n-hexane on the level of sex hormones and expression of estrogen receptor(ER) in rats and the protective effect of Lyciumbarbarum polysaccharide(LBP) on n-hexane-induced reproductive toxicity. METHODS: Based on factorial design model of 4×2, specific pathogen free adult female SD rats were divided into control group and low-, medium-and high-n-hexane exposure groups, and each group was divided into non-LBP intervention and LBP intervention sub-group. There were 8 subgroups with 6 rats in each group. On the first day, the rats in the 4 groups were given intraperitoneal injection of n-hexane at 0, 675, 1 350 and 2 700 mg/kg body weight, respectively. On day 2-4, the rats in the non-LBP intervention subgroup were given intragastric administration of 0.9% sodium chloride solution, and the rats in the LBP intervention subgroup were given intragastric administration of LBP at 50 mg/kg body weight once a day. On the fifth day, all animals were sacrificed, and the levels of follicle stimulating hormone(FSH), luteinizing hormone(LH), estradiol, progesterone were detected by enzyme linked immunosorbent assay. The mRNA expression of Erα, Erβ and G protein coupled estrogen receptor 1(Gper1) was detected by real time fluorescence polymerase chain reaction, and the expression of ERα, ERβ and GPER1 protein was detected by Western blotting. RESULTS: i) In the absence of LBP intervention(i.e. simple n-hexane exposure), there was no significant difference in the level of serum FSH, LH, estradiol and progesterone in the 4 groups(P>0.05). The relative expression of Erβ mRNA in ovary of low dose group decreased, while the relative expression of proteins of ERα and GPER1 increased(P<0.05) when compared with the control group. The relative expression of Erα mRNA and GPER1 protein in the ovary of medium-and high-dose groups increased(P<0.05), while the relative expression of Erβ, Gper1 mRNA and ERβ protein decreased(P<0.05). The relative expression of ERα protein in ovary of high-dose group increased(P<0.05). ii) At the same dose of n-hexane exposure, the relative expression of Erα mRNA in ovary of rats in low dose group increased(P<0.05), while the relative expression of ERβ and GPER1 protein decreased in LBP intervention group compared with the no LBP intervention group(P<0.05). The relative expression of ERα and GPER1 protein in ovary of medium dose group increased(P<0.05), while the relative expression of Gper1 mRNA and GPER1 protein in ovary of high dose group decreased in LBP intervention group compared with the no LBP intervention group(P<0.05). CONCLUSION: n-Hexane can up-regulate the expression of ERα and GPER1 in rat ovary, but has no significant effect on female endocrine system. LBP may play a protective role in female reproductive system by up-regulating the expression of ERα and GPER1.

Objective: To explore the prognostic effects of mean corpuscular volume (MCV) in patients with myelodysplastic syndromes (MDS) . Methods: 321 newly diagnosed, untransfused primary MDS patients who administered from December 2009 to December 2017 were enrolled. The association of MCV with prognosis and several clinical features and genetic mutations were analyzed. Results: Patients were divided into MCV≤100 fl (n=148) and MCV>100 fl (n=173) cohorts. Median overall survival of patients with MCV≤100 fl was shorter than their counterparts (27 months vs 72 months, P<0.001) . In subgroup analysis, MCV≤100 fl patients had worse survivals in bone marrow blast <5% cohort (34 months vs not reached, P=0.002) , but not so in ≥5 % cohort (17 months vs 20 months, P=0.078) . MCV≤100 fl was still an independent adverse variable (HR=1.890, 95%CI 1.007-3.548, P=0.048) after adjusting for clinical and laboratory variables and mutation topography in bone marrow blasts<5% cohort. In bone marrow blasts<5% cohort, patients with MCV≤100 fl had higher hemoglobin levels [90 (42-153) g/L vs 78.5 (28-146) g/L, P=0.015].The proportions of Revised International Prognostic Scoring System (IPSS-R) high/very high risks and poor/very poor IPSS-R karyotypes were higher in MCV≤100 fl cohort (28.8% vs 10.8%, P=0.003; 24.7% vs 12.9%, P=0.049) . MCV≤100 fl cohort had more genetic mutations than those with MCV>100 fl though without significance (0.988 vs 0.769, P=0.064) . Mutated SF3B1 was less frequently in MCV≤100 fl cohort (4.7% vs 15.4%, P=0.018) . Conclusion MCV≤100 fl was an independent adverse variable after adjusting for clinical and laboratory variables and mutation topography in MDS patients with bone marrow blasts<5%.

Objective:To observe the changes of foveal avascular zone (FAZ) size before and after surgery in idiopathic macular epiretinal membrane (IMEM) eyes and analyze the correlation of FAZ with metamorphopsia.Methods:A retrospective case series study. From August 2016 to October 2017, 42 eyes of 38 patients affected with IMEM diagnosed in Central Theater Command General Hospital of Chinese People's Liberation Army were enrolled in this study. All the patients underwent a 25G pars plana vitrectomy (PPV) with IMEM removal and ininternal limiting membrane (ILM) peeling. The BCVA was measured using the international standard visual acuity chart, and the results were converted to the logMAR visual acuity. The severity of metamorphopsia was measured using M-charts. The FAZ areas were evaluated with OCT angiography in both the superficial and deep capillary plexus layers. The central macular thickness (CMT) were assessed with spectral-domain OCT before and after surgery. The logMAR BCVA was 0.61±0.21. The M-score was 0.66±0.38. The CMT of fovea was 337.71±57.63 μm. The FAZ areas in superficial and deep capillary plexus were 0.113±0.037 mm 2 and 0.202±0.03 mm 2, respectively. The differences in BCVA, M-score and FAZ area before and 1, 3, 6, 12 months after surgery were analyzed by ANOVA. The Spearman rank correlation analysis was performed to investigate the relationship between FAZ areas, visual acuity and metamorphopsia. Results:At 12 month after surgery, the FAZ areas in superficial and deep capillary plexus were 0.146±0.021 mm 2 and 0.240±0.019 mm 2, respectively. Compared with baseline, the the FAZ areas in superficial and deep capillary plexus after surgery significantly increased ( F=8.484, 14.346; P<0.001,<0.001). The postoperative logMAR BCVA 0.47±0.19, M-score 0.12±0.22 and CMT 270.60±33.27 μm were significantly improved compared with baseline ( F=5.044, 17.763, 13.545; P=0.001, <0.001, <0.001). The preoperative FAZ area in superficial capillary plexus correlated negatively with preoperative M-score ( r=-0.816, P<0.001); the preoperative FAZ area in deep capillary plexus correlated negatively with preoperative BCVA and M-score ( r=-0.422, -0.882; P=0.005,<0.001). The postoperative FAZ area in superficial capillary plexus correlated negatively with preoperative and postoperative M-score ( r=-0.791,-0.716; P<0.001,<0.001). The postoperative FAZ area in deep capillary plexus correlated negatively with BCVA and preoperative and postoperative M-score ( r=-0.343, -0.330, -0.732, -0.694; P=0.026, 0.033,<0.001,<0.001). Conclusions:PPV with ILM peeling can effectively restore the FAZ areas in superficial and deep capillary plexus, improve the visual acuity and metamorphopsia in patients with IMEM. Both superficial and deep plexus FAZ areas correlated negatively with metamorphopsia, and deep plexus FAZ area also correlated negatively with BCVA.

Objective:To examine the surgical approach, practical cognition as well as clinical effect of the orthotopic resection for laparoscopic pancreatoduodenectomy(OLPD).Methods:From March 2019 to December 2019, 32 cases were treated with laparoscopic pancreatoduodenectomy (LPD) in a novel approach without mobilization of pancreatoduodenum in Pancreas Center of the Second Affiliated Hospital of Guangzhou University of Chinese Medicine.There were 16 male patients and 16 female patients.The mean age was (64.8±9.5) years old.Body mass index was 14.9 to 31.0 kg/m 2.All patients were diagnosed as ampullary or pancreatic head tumors and were not unresectable cases.In the surgical strategy, Kocher′s dissociation, turning and pulling of the pancreaticoduodenal region, was not performed first.Anatomy in situ, separation of vessels which enter and exit from pancreas, separation of lymphatics and isolation of tumors were carried out in priority through the combined middle and left posterior approaches.Finally, the pancreatic head and duodenum region was mobilized and the entire resection of pancreas in situ was carried out.Digestive tract reconstruction was performed through Child method. Results:Postoperative pathology showed that 27 cases were pancreatic or ampullary malignant tumors and five cases were benign tumors among 32 patients.The operative time was (357.3±64.3) minutes.The diameter of pancreatic ducts was (3.0±1.0) mm. The pancreas of 20 cases (62.5%) were soft. Five patients suffered from pancreatic fistula (Grade B) and one patient suffered from intra-abdominal hemorrhage postoperatively.No other complications like pancreatic fistula (Grade C) or biliary fistula delayed gastric emptying or mortality were encountered.The postoperative hospital day was (13.7±3.6) days.Conclusions:Combining the multi-angle of the laparoscopic approaches and excising the pancreaticoduodenal specimen in situ, OLPD is a kind of surgical method which can realize the concept of no touch tumor surgery.Patients who undergo the OLPD can receive better treatments and results.