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.

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%.

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 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.

Objective:To explore the features and clinical significance of gene mutations in patients with myelodysplastic syndromes with ring sideroblasts (MDS-RS) .Methods:A total of 255 newly diagnosed primary MDS-RS patients were retrospectively reviewed from our center from January2001 to June 2019. SF3B1 gene mutations were detected by Sanger sequencing in 129 patients, and next generation sequencing (NGS) was performed in the other 126 patients using a set of selected 112-genes.Results:A total of 193 (75.7%) patients presented with SF3B1 mutation, predominantly mutant at amino acid position 700 (K700E) ( n=147, 76.2%) . Non-SF3B1 gene mutations were TET2 (16.7%) , ASXL1 (14.3%) , U2AF1 (11.1%) , TP53 (7.9%) , SETBP1 (6.3%) , and RUNX1 (6.3%) . RS 5%-<15% patients had a higher SETBP1 mutation frequency than RS≥15% patients (21.4% vs 4.5%, P=0.044) . Mutation frequencies of other genes were similar in both groups (all P>0.05) . SF3B1 variant allele frequencies (VAF) had positive correlation with marrow RS percentage but without statistical significance in RS 5%-<15% group ( P=0.078, r=0.486) . SF3B1 mutant patients presented with higher marrow RS percentage compared with wild-type patients[40.0% (15.0%-80.0%) vs 25.5% (15.0%-82.0%) , P<0.001], and SF3B1 VAF positively correlated with RS percentage ( P=0.009, rs=0.261) in RS≥15% group. Age, ANC, PLT, mean RBC corpuscular volume, RS percentage, IPSS-R cytogenetics, and IPSS-R risk score were significantly different between patients with SF3B1 mutations and wild-type SF3B1 (all P<0.05) . Multivariable survival analyses adjusted by age and IPSS-R cytogenetics revealed that SF3B1 mutation was an independent favorable prognostic factor ( HR=0.265, 95% CI 0.077-0.917, P=0.036) , and TP53 mutation was an adverse variable independent of SF3B1 mutation ( HR=6.272, 95% CI 1.725-22.809, P=0.005) . According to the mutant status of SF3B1 and TP53, MDS-RS patients were categorized into 4 groups, namely, with SF3B1 and TP53 mutation, with wild-type SF3B1 and TP53, with wild-type SF3B1 but TP53 mutation, and with SF3B1 mutation but wild-type TP53. There was a significant difference for OS among these 4 groups ( P<0.001) . The former 3 groups showed no significant difference in OS in multiple comparisons. However, the SF3B1 mutation but wild-type TP53 group had a better OS than wild-type SF3B1 but TP53 mutation group and wild-type SF3B1 and TP53 group, whereas a similar OS compared with SF3B1 and TP53 mutation group. Conclusion:SF3B1 mutations were prevalent in MDS-RS patients with the most common mutation at amino acid position 700 (K700E) . SF3B1 mutation was an independent favorable prognostic variable, whereas TP53 mutation was an independent adverse variable. SF3B1 mutation could coordinate with TP53 mutation for more sophisticated prognosis stratification in MDS-RS patients.

Objective:To compare fibrosis-driving cells in patients with primary myelofibrosis (PMF) and patients with myelodysplastic syndromes (MDS) with myelofibrosis (MF) (MDS-MF) .Methods:Bone marrow biopsy sections of patients with newly diagnosed PMF and MDS (10 each randomly selected for MF-0/1, MF-2, and MF-3) were stained with specific immunofluorescence antibodies to label Gli1, LeptinR, alpha smooth muscle actin (α-SMA) , CD45, and ProcollagenⅠ. Images captured by confocal microscopy were analyzed by Fiji-ImageJ to calculate the cell counts of Gli1 +, LeptinR + cells, and fibrosis-driving cells including α-SMA +, α-SMA +/Gli1 +, α-SMA +/LeptinR +, and ProcollagenⅠ +/CD45 + cells. Results:Patients with PMF and MDS with MF-2/3 had higher LeptinR +, α-SMA +, α-SMA +/Gli1 +, and Procollagen Ⅰ +/CD45 + cell counts compared with those with MF-0/1 (all P values<0.05) . However, patients with PMF with MF-2/3 presented with higher Gli1 + and α-SMA +/LeptinR + cell counts than those with MF-0/1 ( P=0.001 and 0.006) , whereas these cells were similar between patients with MDS with MF-0/1 and MF-2/3 ( P=0.169 and 0.067) . In patients with MF-0/1, all fibrosis-driving cells did not differ between PMF and MDS (all P>0.05) . However, in patients with MF-2/3, Procollagen Ⅰ +/CD45 + cell counts were higher in patients with PMF compared with those with MDS ( P=0.007) , while other fibrosis-driving cell counts were similar between these two groups (all P>0.05) . MF grade and fibrosis-driving cell counts were not correlated with overall survival in patients with either PMF or MDS. Conclusion:α-SMA + cells in patients with PMF originated from both Gli1 + and LeptinR + cells, whereas α-SMA + cells in patients with MDS-MF only originated from Gli1 + cells; patients with PMF had higher ProcollagenⅠ +/CD45 + cell counts than those with MDS-MF.

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.

Objective: To explore the clinical implications and prognostic value of TP53 gene mutation and deletion in patients with myelodysplastic syndromes (MDS) . Methods: 112-gene targeted sequencing and interphase fluorescence in situ hybridization (FISH) were used to detect TP53 mutation and deletion in 584 patients with newly diagnosed primary MDS who were admitted from October 2009 to December 2017. The association of TP53 mutation and deletion with several clinical features and their prognostic significance were analyzed. Results: Alterations in TP53 were found in 42 (7.2%) cases. Of these, 31 (5.3%) cases showed TP53 mutation only, 8 (1.4%) cases in TP53 deletion only, 3 (0.5%) cases harboring both mutation and deletion. A total of 37 mutations were detected in 34 patients, most of them (94.6%) were located in the DNA binding domain (exon5-8) , the remaining 2 were located in exon 10 and splice site respectively. Patients with TP53 alterations harbored significantly more mutations than whom without alterations (z=-2.418, P=0.016) . The median age of patients with TP53 alterations was higher than their counterparts[60 (21-78) years old vs 52 (14-83) years old, z=-2.188, P=0.029]. TP53 alterations correlated with complex karyotype and International prognostic scoring system intermediate-2/high significantly (P<0.001) . Median overall survival of patients with TP53 alterations was shorter than the others[13 (95%CI 7.57-18.43) months vs not reached, χ²=12.342, P<0.001], while the significance was lost during complex karyotype adjusted analysis in multivariable model. Conclusion TP53 mutation was more common than deletion in MDS patients. The majority of mutations were located in the DNA binding domain. TP53 alterations were strongly associated with complex karyotype and always coexisted with other gene mutations. TP53 alteration was no longer an independent prognostic factor when complex karyotype were occurred in MDS.