BACKGROUND:Spleen has the functions of blood storage,hematopoiesis,and immunity.With the increase of age,the structural degeneration and functional decline of spleen lead to the impairment of immune system function,thus accelerating the aging process of the body.The treatment of spleen aging in tree shrews with highly active umbilical cord mesenchymal stem cells has not been reported. OBJECTIVE:To explore the intervention effect and mechanism of highly active umbilical cord mesenchymal stem cells on spleen aging in tree shrews. METHODS:Highly active umbilical cord mesenchymal stem cells were isolated,cultured,and obtained from the umbilical cord tissue of newborn tree shrews by caesarean section.The differentiation abilities of adipogenesis,osteogenesis,and chondrogenesis were detected by three-line differentiation kit.Cell cycle and surface markers were detected by flow cytometry.The second generation of highly active umbilical cord mesenchymal stem cells were transfected with Genechem Green Fluorescent Protein with infection complex values of 100,120,140,160,180,and 200,respectively,to screen the best transfection conditions.After transfection,the fourth generation of highly active umbilical cord mesenchymal stem cells was injected into the tail vein of tree shrews in the elderly treatment group.The young control group and the aged model group were not given special treatment.After 4 months of treatment,the spleen tissue was taken and the structure of the spleen was observed by hematoxylin-eosin staining.β-Galactosidase staining was used to detect the activity of aging-related galactosidase.Immunohistochemical staining was used to detect the expression levels of p21 and p53 proteins.Ki67 and PCNA immunofluorescence staining was used to detect cell proliferation activity.Immunofluorescence staining was used to detect the expression levels of spleen autophagy protein molecules Beclin 1 and APG5L/ATG5.Reactive oxygen species fluorescence staining was used to detect the content of reactive oxygen species in spleen tissue.CD3 immunofluorescence staining was used to detect the change of the proportion of total T lymphocytes.The secretion levels of interleukin 1β and transforming growth factor β1 in spleen were detected by enzyme linked immunosorbent assay.The distribution of highly active umbilical cord mesenchymal stem cells labeled with green fluorescent protein in spleen tissue was observed by DAPI double staining of nucleus. RESULTS AND CONCLUSION:(1)Highly active umbilical cord mesenchymal stem cells grew in a short spindle shape with fish-like growth,with a large proportion of G0/G1 phase,and had the potential to differentiate into adipogenesis,osteogenesis,and chondrogenesis.(2)Multiplicity of infection=140 and transfection for 72 hours were the best conditions for labeling tree shrews highly active umbilical cord mesenchymal stem cells with Genechem Green Fluorescent Protein.(3)Compared with the aged model group,in the aged treatment group,the spleen tissue cells of tree shrews were arranged closely,and the area of white pulp was increased(P<0.01);the boundary between red pulp and white pulp was clear;the proportion of germinal centers did not show statistically significant difference(P>0.05).The activity level of galactosidase related to spleen tissue aging was decreased(P<0.001),and the expression levels of aging protein molecules p21 and p53 were down-regulated(P<0.001).The expression levels of proliferation-related molecules Ki67 and PCNA were up-regulated(P<0.001,P<0.05);expression levels of autophagy-related molecules Beclin 1 and APG5L/ATG5 were up-regulated(P<0.001),and the content of reactive oxygen species decreased(P<0.001),and the proportion of CD3+T cells increased(P<0.05).The secretion level of interleukin 1β in the aging-related secretion phenotype decreased(P<0.001);no significant difference was found in transforming growth factor β1 level(P>0.05).Compared with the young control group,the above indexes were significantly different in the elderly treatment group(P<0.05).(4)Green fluorescent cells labeled with green fluorescent protein were observed in spleen tissue of tree shrews the elderly treatment group by frozen tissue section observation.The results show that intravenous infusion of highly active umbilical cord mesenchymal stem cells can migrate to spleen tissue,inhibit the production of reactive oxygen species,down-regulate the expression of aging-related proteins,induce autophagy,promote cell proliferation,reduce chronic inflammation,and then improve the structure and function of spleen tissue.

Objective:To observe and evaluate the clinical efficacy and safety of albumin-bound paclitaxel as first-line treatment for patients with advanced pancreatic cancer in China, and to explore the prognosis-related molecules in pancreatic cancer based on next-generation sequencing (NGS) of tumor tissues.Methods:From December 2018 to December 2020, patients with locally advanced or metastatic pancreatic cancer were recruited to accept albumin-bound paclitaxel as first-line treatment in the oncology departments of 24 hospitals in East China. The primary endpoints were overall survival (OS) and treatment related adverse events, and the secondary endpoint was progression-free survival (PFS). Adverse effects were graded using Common Terminology Criteria for Adverse Events 5.0 (CTCAE 5.0). NGS sequencing on the primary or metastatic tissue samples of pancreatic cancer obtained through surgical resection or biopsy was performed.Results:This study recruited 229 patients, including 70 patients with locally advanced pancreatic cancer (LAPC) and 159 patients with metastatic pancreatic cancer (mPC). The disease control rate was 79.9% and the objective response rate is 36.3%.The common adverse effects during treatment were anaemia (159 cases), leucopenia (170 cases), neutropenia (169 cases), increased aminotransferases (110 cases), and thrombocytopenia (95 cases), and the incidence of grade 3-4 neutropenia is 12.2% (28/229). The median follow-up time was 21.2 months (95% CI: 18.5-23.1 months). The median PFS (mPFS) was 5.3 months (95% CI: 4.37-4.07 months) and the median OS (mOS) was 11.2 months (95% CI: 9.5-12.9 months). The mPFS of patients with LAPC was 7.4 months (95% CI: 6.6-11.2 months), and their mOS was 15.5 months (95% CI: 12.6-NA months). The mPFS of patients with mPC was 3.9 months (95% CI: 3.4-5.1 months), and their mOS was 9.3 months (95% CI: 8.0-10.8 months). Multivariate Cox regression analysis showed that clinical stage ( HR=1.47, 95% CI: 1.06-2.04), primary tumor site ( HR=0.64, 95% CI: 0.48-0.86), Eastern Cooperative Oncology Group Performance Status (ECOG PS) score ( HR=2.66, 95% CI: 1.53-4.65), and whether to combine radiotherapy ( HR=0.65, 95% CI: 0.42-1.00) were independent influencing factors for the PFS of these patients. The primary tumor site ( HR=0.68, 95% CI: 0.48-0.95), ECOG score ( HR=5.82, 95% CI: 3.14-10.82), and whether to combine radiotherapy ( HR=0.58, 95% CI: 0.35-0.96) were independent influencing factors of the OS of these patients. The most frequent gene mutations in these advanced stage pancreatic patients were KRAS (89.66%), TP53 (77.01%), CDKN2A (32.18%), and SMAD4 (21.84%) by NGS of tumor tissues from 87 pancreatic cancer patients with sufficient specimens. Further analysis revealed that mutations in CDKN2B, PTEN, FGF6, and RBBP8 genes were significantly associated with an increased risk of death ( P＜0.05). Conclusion:Albumin-bound paclitaxel as first-line treatment demonstrated feasible anti-tumor efficacy and manageable safety for patients with advanced pancreatic cancer in China.

Objective:To observe and evaluate the clinical efficacy and safety of albumin-bound paclitaxel as first-line treatment for patients with advanced pancreatic cancer in China, and to explore the prognosis-related molecules in pancreatic cancer based on next-generation sequencing (NGS) of tumor tissues.Methods:From December 2018 to December 2020, patients with locally advanced or metastatic pancreatic cancer were recruited to accept albumin-bound paclitaxel as first-line treatment in the oncology departments of 24 hospitals in East China. The primary endpoints were overall survival (OS) and treatment related adverse events, and the secondary endpoint was progression-free survival (PFS). Adverse effects were graded using Common Terminology Criteria for Adverse Events 5.0 (CTCAE 5.0). NGS sequencing on the primary or metastatic tissue samples of pancreatic cancer obtained through surgical resection or biopsy was performed.Results:This study recruited 229 patients, including 70 patients with locally advanced pancreatic cancer (LAPC) and 159 patients with metastatic pancreatic cancer (mPC). The disease control rate was 79.9% and the objective response rate is 36.3%.The common adverse effects during treatment were anaemia (159 cases), leucopenia (170 cases), neutropenia (169 cases), increased aminotransferases (110 cases), and thrombocytopenia (95 cases), and the incidence of grade 3-4 neutropenia is 12.2% (28/229). The median follow-up time was 21.2 months (95% CI: 18.5-23.1 months). The median PFS (mPFS) was 5.3 months (95% CI: 4.37-4.07 months) and the median OS (mOS) was 11.2 months (95% CI: 9.5-12.9 months). The mPFS of patients with LAPC was 7.4 months (95% CI: 6.6-11.2 months), and their mOS was 15.5 months (95% CI: 12.6-NA months). The mPFS of patients with mPC was 3.9 months (95% CI: 3.4-5.1 months), and their mOS was 9.3 months (95% CI: 8.0-10.8 months). Multivariate Cox regression analysis showed that clinical stage ( HR=1.47, 95% CI: 1.06-2.04), primary tumor site ( HR=0.64, 95% CI: 0.48-0.86), Eastern Cooperative Oncology Group Performance Status (ECOG PS) score ( HR=2.66, 95% CI: 1.53-4.65), and whether to combine radiotherapy ( HR=0.65, 95% CI: 0.42-1.00) were independent influencing factors for the PFS of these patients. The primary tumor site ( HR=0.68, 95% CI: 0.48-0.95), ECOG score ( HR=5.82, 95% CI: 3.14-10.82), and whether to combine radiotherapy ( HR=0.58, 95% CI: 0.35-0.96) were independent influencing factors of the OS of these patients. The most frequent gene mutations in these advanced stage pancreatic patients were KRAS (89.66%), TP53 (77.01%), CDKN2A (32.18%), and SMAD4 (21.84%) by NGS of tumor tissues from 87 pancreatic cancer patients with sufficient specimens. Further analysis revealed that mutations in CDKN2B, PTEN, FGF6, and RBBP8 genes were significantly associated with an increased risk of death ( P＜0.05). Conclusion:Albumin-bound paclitaxel as first-line treatment demonstrated feasible anti-tumor efficacy and manageable safety for patients with advanced pancreatic cancer in China.

Objective To analyze the clinical characteristics,diagnosis,treatment and prognostic factors of primary hepatic neuroendocrine tumor (PHNET).Methods To analyze the clinical data of 21 patients with PHNET who were admitted to the First Affiliated Hospital of Zhengzhou University from January 2012 to January 2019.There were 11 males and 10 females,with ages which ranged from 36 to 75 years.Log-rank test was used for single-factor analysis of the relationship between clinical and pathological indicators and prognosis.Cox proportional risk model was used for multi-factor analysis.Results Among the 21 PHNET patients,10 presented with central and upper abdominal pain,3 abdominal pain accompanied by nausea and vomiting,5 abdominal distension,and 3 were asymptomatic.Carbohydrate antigen (CA) 19-9 was elevated in 6 patients and CA125 in 7 patients.Abdominal enhanced CT showed solid liver space occupying lesions.Four patients had solitary and 17 had multiple lesions.The mean diameter was 58 mm.Fourteen patients had lymph node metastasis.Five patients underwent radical hepatectomy,2 transcatheter hepatic arterial chemoembolization + chemotherapy,12 chemotherapy,and 2 supportive treatment.Factors which were associated with prognosis of PHNET patients were surgery,tumor grading,cytokeratin positivity and lymph node metastasis.Multivariate Cox regression analysis showed that inoperability (HR =8.99,95％ CI:1.13-71.80) was an independent risk factor of prognosis of PHNET patients.The prognosis in patients who underwent surgical resection was better.Conclusion Patients with PHNET had no specific clinical manifestations.Surgical resection gave the best results in treatment.As surgical resection affected prognosis,it should be carried out if technically feasible.

Objective To study the effects of two kinds of vertical partial laryngectomy on laryngeal function,postoperative extubation rate and survival rate of patients with glottic laryngeal carcinoma.Methods From January 2013 to January 2014,one hundred and fifty patients with glottic carcinoma who treated in Huzhou Gospel Hospital were selected.The patients were divided into observation group and control group by random number table method,with 75 cases in each group.The control group used the traditional vertical partial laryngectomy,the observation group was treated with modified vertical partial laryngectomy.The postoperative survival rate,extubation rate,extubation time and complications were observed in the two groups.Results The extubation rate was 100.00% (75/75) in the observation group and 97.33% (73/75) in the control group,the difference was statistically significant between the two groups(χ2 =2.027,P>0.05).The extubation time in the observation group was (11.85 ± 0.49)d,which in the control group was (14.55 ± 0.56) d,the difference was statistically significant between the two groups( t=31.424, P<0.05).There were 67 cases(89.33% ) with grade 0 and 8 cases(10.67% ) with grade 1 in the observation group.The swallowing function of the observation group was stronger than that of the control group( Z=5.238,P<0.05).The 1-year and 2-year survival rates of the two groups were similar(all P>0.05).The 3-year survival rate of the observation group was 97.33% (73/75),which of the control group was only 88.00% (66/75),the differ-ence was statistically significant between the two groups (χ2 =4.807,P<0.05).There was no pharyngeal fistula in both two groups. There were 5 incision infections in the control group, and 1 incision infection in the observation group.There was no statistically significant difference in the incidence rate of complications between the two groups (χ2 =2.778,P>0.05).Conclusion Improved vertical hemilaryngectomy therapy for patients with glottic laryngeal carcinoma can effectively shorten the time of extubation,better preserve swallowing function,improve the long-term survival rate,and the extubation rate and complication rate are similar with the traditional vertical partial laryngectomy.

Objective This study aimed to evaluate the impact of Intensive Care Unit (ICU)admission on patients with malignant liver tumors after elective major hepatectomy,and to analyze the relevant factors relating to ICU admission.Method 1 044 liver cancer patients who underwent elective hepatectomy were included into this study.Using the length of extubation time (＞ 5 h) and/or treatment time (＞ 24 h) in ICU,the patients were divided into the ICU group and the general group.The age,gender,underlying disease and operative time,intraoperative blood loss,blood transfusion,as well as postoperative complications,mortality,hospital stay and hospitalization costs were compared between the two groups.The relative factors of the ICU group were evaluated using univariate and multivariate analysis.Results Compared to the general group,the mean age (57.4 ± 3.7 vs.53.2 ± 3.2),and the proportions of patients with associated lung diseases (33.7％ vs.10.0％),chronic kidney diseases (9.6％ vs 2.0％),anemia (50.6％ vs.5.3％),as well as the amounts of intraoperative blood loss (816 ml vs.635 ml) and blood transfusion (4.3 U vs.1.4 U) in the ICU group were significantly higher.The differences were significant (P ＜0.05).Furthermore,the ICU group of patients had a significantly longer hospital stay (21.6 days vs.10.1 days,P ＜ 0.05) and more hospital costs (76 751 yuan vs.42 069 yuan,P ＜ 0.05).Multivariate analysis showed that age,blood loss and transfusion were associated with ICU admission.Conclusions It is not necessary to admit every patient with malignant liver tumors to ICU after elective major hepatectomy.ICU admission resulted in prolonged hospital stay and elevated hospital costs.Age (OR =1.077,95％ CI:1.030 ～ 1.127),amount of blood loss (OR =3.211,95％ CI:1.038 ～ 9.929) and blood transfusion (OR =1.330,95％ CI:1.113 ～ 1.589) were associated with ICU admission.There are still many potentially influencing factors which need further studies to determine.

Objective To explore the clinical significance of intraoperative frozen sections for the diagnosis of unexpected gallbladder neoplasm during cholecystectomy for acute cholecystitis.Methods We retrospectively analyzed the clinical data of acute cholecystitis patients who underwent cholecystectomy at the First Affiliated Hospital of Zhengzhou University,from Dec 2012 to Dec 2017.Results In the 1 386 acute cholecystitis patients,19 patients were found to have concurrent gallbladder neoplasm.Surgeons accurately recognized 9 gallbladder neoplasms by general observation alone,including 2 T2 and 2 T3 gallbladder neoplasms,but missed 10 gallbladder neoplasms.At the same time,we found that 3 Tis and 1 T1a gallbladder neoplasms were missed by frozen sections from 10 gallbladder neoplasms diagnosed by definitive histopathological examination.The sensitivity of frozen sections diagnosis was 60％ and the specificity was 100％.Conclusions During cholecystectomy for acute cholecystitis,the accuracy of surgeons' diagnosis with general observation in unexpected gallbladder neoplasm is poor.The accuracy of frozen sections to diagnose advanced gallbladder neoplasm is high,so we advocate frozen sections of every cholecystectomy sample in acute cholecystitis patients.

Objective To investigate the role of miR-200b on gemcitabine induced epithelialmesenchymal transition (EMT) in pancreatic cancer cell line MiaPaCa-2.Methods Different concentrations of gemcitabine were used to induce MiaPaCa-2,and the concentration of 50％ cell proliferation inhibited (IC50) was applied to obtain drug-resistant MiaPaCa-2 cells.MiR-200b or nonsense small molecular fragments (negative control,NC) was transfected into MiaPaCa-2 cells by liposomes,then gemcitabine of IC50 was used to induce cells to obtain drug-resistant MiaPaCa-2 cells transfected with miR-200b or NC.The morphological characteristics of MiaPaCa-2 cells were observed by inverted microscope.Invasion of cells were detected by transwell chamber.The expression of miR-200b was measured by using real-time PCR.The expressions of Ecadherin,Vimentin,Zebl,Zeb2 proteins were determined by Western blot.Results After gemcitabine treatment,the cells' size gradually diminished,intercellular junctions decreased,pseudopodium increased,which presented the characteristics of mesenchymal morphology.The invaded cell number increased from (26 ± 3) to (85 ± 6),and the expression of Vimentin Zebl,Zeb2 was increased to (1.87 ± 0.17),(2.57 ±0.21),(5.24 ± 0.83) folds of the parent cells.The expression of miR-200b was decreased to (0.36 ± 0.01)folds of the parent cells,and the expression of E-cadherin was decreased to 0.47 ± 0.05 folds of the parent cells,while the invaded cell number of drug-resistant MiaPaCa-2 transfected with miR-200b was decreased to (42 ± 4),and the expression of Zebl,Zeb2 was decreased to (0.36 ± 0.07),(0.08 ± 0.01) folds of drugresistant MiaPaCa-2 transfected with NC.Conclusions The occurrence of EMT is observed in pancreatic cancer cell line MiaPaCa-2 during gemcitabine induction,and miR-200b down-regulation may be a possible mechanism.

Objective To summarize the experience in the diagnosis and treatment of solid pseudopapillary tumors of the pancreas. Methods Ten consecutive patients who underwent surgery with pathologically confirmed solid pseudopapillary tumors of the pancreas between October 2005 and December 2008 were retrospectively reviewed. Results All of the 10 patients were female and the median age at diagnosis was 24 years (range, 11 -39 years). Abdominal discomfort or pain were the most common presenting symptoms. 4 patients had palpable abdominal mass at physical examination. The tumors appeared on ultrasonography and/or CT, MRI as solid or cystic masses. The preoperative serum biochemical parameters and tumor markers level were within the normal range. All the patients underwent surgical treatment. The tumors were located in the head/neck (n = 6) or the distal part (n = 4) of the pancreas. The surgical procedures included enucleation (n=3) , distal pancreatectomy (n=3 , two with preservation of the spleen, one combined with splenectomy, distal gastrectomy and partial colectomy) , segmental pancreatectomy with pancreaticojejunostomy (n=3) and pancreaticoduodenectomy (n = 1). Pancreatic fistula (n = 2) was observed postoperatively and resolved with conservative treatment. The median resected tumor size was 5. 9 cm. All patients were alive and remained recurrence and metastasis free after a median followk-up of 19. 2 months (range, 8～42 months). Conclusions Solid pseudopapillary tumor of the pancreas was rare neoplasm occurred predominantly in young women with low malignant potential. Aggressive resection should be attempted and could result in excellent prognosis.

Objective To investigate the clinical features, diagnosis and treatment of malignant pancreatic endocrine tumor. Methods The clinical data of 38 patients with malignant pancreatic endocrine tumor who had been admitted to First Affiliated Hospital of Nanjing Medical University from January 1969 to December 2008 were analyzed retrospectively. Of all patients, 6 were with insulinoma, 23 with pancreatic polypeptide tumor, 4 with glucagonoma and 5 with pancreatic carcinoid. Results All patients except 1 with insulinoma were found with pancreatic lesion by imaging examination. The resection rate was 87% (33/38). Pathological examination found 7 patients with liver metastasis, 5 with lymph node metastasis, 1 with tumor thrombus in vessels and lymphatic vessels, and 28 with local invasion. Twenty-four patients were followed up, and neither recurrence nor metastasis was found except 1 patient with insulinoma who received reoperation for local recurrence and 1 patient with pancreatic carcinoid who received radiofrequency ablation for liver metastasis. Conclusions The diagnosis of pancreatic endocrine tumor mainly depends on imaging examination. The malignancy of pancreatic endocrine tumor is determined after the comprehensive analysis of preoperative imaging findings, intraoperative examination, post-operative pathological examination and the data obtained during follow-up. The malignant pancreatic endocrine tumor should be managed actively by resection because of its relatively low malignancy, high operative resectability and relatively good prognosis.