Characterization and phylogenetic analysis of chloroplast genome of Cynanchum wallichii and Cynanchum otophyllum / 药学学报

italic>Cynanchum wallichii and Cynanchum otophyllum belong to the genus Cynanchum in the family Apocynaceae, and are important medicinal plants. In this study, we sequenced and assembled the chloroplast genomes of C. wallichii and C. otophyllum, and performed a phylogenetic analysis of the structural characteristics of their chloroplast genomes and their phylogenetic positions. The results showed that the chloroplast genomes of both C. wallichii and C. otophyllum had a typical tetrad structure, with 133 genes annotated, and the total GC contents of both were similar. Codon preference analysis showed that the relative synonymous codon usage in the chloroplast genomes of C. wallichii and C. otophyllum differed slightly, but the differences were not significant, and there was a strong A or U preference at the third codon position. In both chloroplast genomes, 91 and 103 simple sequence repeats were detected respectively, and the largest proportion of A/T type repeats. Nucleotide polymorphism analysis showed that the nucleotide diversity of the intergenic sequences in the chloroplast genome of genus Cynanchum were generally higher than those of the common gene sequences. A pair of primers was designed based on the high variation region of the chloroplast genome to identify C. wallichii and C. otophyllum. The phylogenetic analysis showed that the C. wallichii and Cynanchum thesioides were the closest relatives, while the C. otophyllum, Cynanchum bungei and Cynanchum wilfordii formed a stable monophyletic clade within the genus Cynanchum, and the three species were closely related. The comparative analysis of the chloroplast genomic characteristics and phylogeny of C. wallichii and C. otophyllum will provide a theoretical basis for the species identification of the two plants and for the study of genetic diversity and phylogeny of the genus Cynanchum.

A nomogram for preoperative prediction of lymph node metastasis in patients with intrahepatic cholangiocarcinoma based on inflammation-related markers / 中华外科杂志

Objectives: To construct a nomogram for prediction of intrahepatic cholangiocarcinoma (ICC) lymph node metastasis based on inflammation-related markers,and to conduct its clinical verification. Methods: Clinical and pathological data of 858 ICC patients who underwent radical resection were retrospectively collected at 10 domestic tertiary hospitals in China from January 2010 to December 2018. Among the 508 patients who underwent lymph node dissection,207 cases had complete variable clinical data for constructing the nomogram,including 84 males,123 females,109 patients≥60 years old,98 patients<60 years old and 69 patients were pathologically diagnosed with positive lymph nodes after surgery. Receiver operating characteristic curve was drawn to calculate the accuracy of preoperative imaging examinations to determine lymph node status,and the difference in overall survival time was compared by Log-rank test. Partial regression squares and statistically significant preoperative variables were screened by backward stepwise regression analysis. R software was applied to construct a nomogram,clinical decision curve and clinical influence curve,and Bootstrap method was used for internal verification. Moreover,retrospectively collecting clinical information of 107 ICC patients with intraoperative lymph node dissection admitted to 9 tertiary hospitals in China from January 2019 to June 2021 was for external verification to verify the accuracy of the nomogram. 80 patients with complete clinical data but without lymph node dissection were divided into lymph node metastasis high-risk group and low-risk group according to the score of the nomogram among the 858 patients. Log-rank test was used to compare the overall survival of patients with or without lymph node metastasis diagnosed by pathology. Results: The area under the curve of preoperative imaging examinations for lymph node status assessment of 440 patients was 0.615,with a false negative rate of 62.8% (113/180) and a false positive rate of 14.2% (37/260). The median survival time of 207 patients used to construct a nomogram with positive or negative postoperative pathological lymph node metastases was 18.5 months and 27.1 months,respectively (P<0.05). Five variables related to lymph node metastasis were screened out by backward stepwise regression analysis,which were combined calculi,neutrophil/lymphocyte ratio,albumin,liver capsule invasion and systemic immune inflammation index,according to which a nomogram was constructed with concordance index(C-index) of 0.737 (95%CI: 0.667 to 0.806). The C-index of external verification was 0.674 (95%CI:0.569 to 0.779). The calibration prediction curve was in good agreement with the reference curve. The results of the clinical decision curve showed that when the risk threshold of high lymph node metastasis in the nomogram was set to about 0.32,the maximum net benefit could be obtained by 0.11,and the cost/benefit ratio was 1∶2. The results of clinical influence curve showed that when the risk threshold of high lymph node metastasis in the nomogram was set to about 0.6,the probability of correctly predicting lymph node metastasis could reach more than 90%. There was no significant difference in overall survival time between patients with high/low risk of lymph node metastasis assessed by the nomogram and those with pathologically confirmed lymph node metastasis or without lymph node metastasis (Log-rank test:P=0.082 and 0.510,respectively). Conclusion: The prediction accuracy of preoperative nomogram for ICC lymph node metastasis based on inflammation-related markers is satisfactory,which can be used as a supplementary method for preoperative diagnosis of lymph node metastasis and is helpful for clinicians to make personalized decision of lymph node dissection for patients with ICC.

Survival analysis of patients with intrahepatic cholangiocarcinoma treated with adjuvant chemotherapy after radical resection based on CoxPH model and deep learning algorithm / 中华外科杂志

Objective: To establish a predictive model for survival benefit of patients with intrahepatic cholangiocarcinoma (ICC) who received adjuvant chemotherapy after radical resection. Methods: The clinical and pathological data of 249 patients with ICC who underwent radical resection and adjuvant chemotherapy at 8 hospitals in China from January 2010 to December 2018 were retrospectively collected. There were 121 males and 128 females,with 88 cases>60 years old and 161 cases≤60 years old. Feature selection was performed by univariate and multivariate Cox regression analysis. Overall survival time and survival status were used as outcome indicators,then target clinical features were selected. Patients were stratified into high-risk group and low-risk group,survival differences between the two groups were analyzed. Using the selected clinical features, the traditional CoxPH model and deep learning DeepSurv survival prediction model were constructed, and the performance of the models were evaluated according to concordance index(C-index). Results: Portal vein invasion, carcinoembryonic antigen>5 μg/L,abnormal lymphocyte count, low grade tumor pathological differentiation and positive lymph nodes>0 were independent adverse prognostic factors for overall survival in 249 patients with adjuvant chemotherapy after radical resection (all P<0.05). The survival benefit of adjuvant chemotherapy in the high-risk group was significantly lower than that in the low-risk group (P<0.05). Using the above five features, the traditional CoxPH model and the deep learning DeepSurv survival prediction model were constructed. The C-index values of the training set were 0.687 and 0.770, and the C-index values of the test set were 0.606 and 0.763,respectively. Conclusion: Compared with the traditional Cox model, the DeepSurv model can more accurately predict the survival probability of patients with ICC undergoing adjuvant chemotherapy at a certain time point, and more accurately judge the survival benefit of adjuvant chemotherapy.

Impact of adjuvant chemotherapy on prognosis in intrahepatic cholangiocarcinoma patients underwent radical resection / 中华外科杂志

Objectives: To investigate the clinical value of adjuvant chemotherapy(ACT) in patients with intrahepatic cholangiocarcinoma(ICC) who underwent radical resection and to explore the optimal population that can benefit from ACT. Methods: A retrospective cohort study method was adopted. The clinical and pathological data of 685 patients with ICC who underwent curative intent resection in 10 Chinese hepatobiliary surgery centers from January 2010 to December 2018 were collected;There were 355 males and 330 females. The age(M(IQR)) was 58(14) years (range: 22 to 83 years). Propensity score matching(PSM) was applied to balance the differences between the adjuvant and non-adjuvant chemotherapy groups. Log-rank test was used to compare the prognosis of the two groups of patients. A Bayesian network recurrence-free survival(RFS) prediction model was constructed using the median RFS time (14 months) as the target variable, and the importance of the relevant prognostic factors was ranked according to the multistate Birnbaum importance calculation. A survival prognostic prediction table was established to analyze the population benefiting from adjuvant chemotherapy. Results: Among 685 patients,214 received ACT and 471 did not receive ACT. A total of 124 pairs of patients were included after PSM, and patients in the ACT group had better overall survival (OS) and RFS than those in the non-ACT group(OS: 32.2 months vs. 18.0 months,P=0.003;RFS:18.0 months vs. 10.0 months,P=0.001). The area under the curve of the Bayesian network RFS prediction model was 0.7124. The results of the prognostic factors in order of importance were microvascular invasion (0.158 2),perineural invasion (0.158 2),N stage (0.155 8),T stage (0.120 9), hepatic envelope invasion (0.090 3),adjuvant chemotherapy (0.072 1), tumor location (0.057 5), age (0.042 3), pathological differentiation (0.034 0), sex (0.029 3), alpha-fetoprotein (0.028 9) and preoperative jaundice (0.008 5). A survival prediction table based on the variables with importance greater than 0.1 (microvascular invasion,perineural invasion,N stage,T staging) and ACT showed that all patients benefited from ACT (increase in the probability of RFS≥14 months from 2.21% to 7.68%), with a more significant increase in the probability of RFS≥14 months after ACT in early-stage patients. Conclusion: ACT after radical resection in patients with ICC significantly prolongs the OS and RFS of patients, and the benefit of ACT is greater in early patients.

The Clinical Evaluation of Danning Tablets on Regulating Gut Flora Displacement to Eliminate Biliary Inflammation and Prevent Gallstone Formation / 昆明医科大学学报

Objective To explore the mechanism of Danning Tablet on intestinal flora migration to eliminate biliary inflammation and prevent gallstone formation. Methods 104 cases of cholelithiasis admitted to the First Affiliated Hospital of Xi'an Jiaotong University School of Medicine were treated by choledocholithiasis but the master tube was cut and T tube drainage was performed from January 2014 to December 2016. Within a month did not take gallbladder drugs. Randomly divided into the Danning group and the control group, Danning group in addition to conventional treatment of biliary tract after surgery, in the first 2 days after starting to drink liquid according to the instruction manual dose of oral Danning tablets;the control group was given only biliary surgery After routine care, avoid taking gall bladder drugs. All patients were collected on the 7th day and the 14 th day after operation, about 100mL fasting T-tube was collected. Pay attention to the day before the collection of bile T tube pre-clamp to prevent the drainage of T tube affect the bile enterohepatic circulation. Detection of bile composition in the flora, observed before and after the flora differences, long-term follow-up of cholelithiasis recurrence. Results:The changes of the components before and after treatment:On the seventh day, the two groups of HS and UCB％were higher than the control group, the difference was statistically significant (<0.05) . On the 14th day in the treatment group, the three items of TBS, UCB％and HS were higher than the control group, but CH, LI and Z were lower than the control group, the difference was statistically significant ( <0.05) . The trend of stone formation before and after treatment:Compared with UCB％in control group, there was no significant change in LI and Z, but the HS value decreased, the difference was statistically significant ( <0.05) . There was no significant change of HS and UCB％in the treatment group, but the LI and Z values decreased, the difference was statistically significant (<0.05) . Conclusion Dan Ning Tablets can prevent cholesterol gallstone formation by regulating the intestinal flora, and affect the cholesterol content in bile , that plays a important role in prevention of cholesterol gallstone.

Healing of stoma after magnetic biliary-enteric anastomosis in canine peritonitis models / 中国医学科学杂志(英文版)

<p><b>OBJECTIVE</b>To assese the healing of stoma after magnetic anastomosis for the reconstruction of biliary-enteric continuity under severe inflammation.</p><p><b>METHODS</b>Acute bile duct injury was constructed as a bile peritonitis model in mongrel dogs (n=32). Magnetic anastomosis (group A, n=16) and traditional suture anastomosis (group B, n=16) were performed to reconstruct the biliary-enteric continuity in one stage. Half of the dogs in each group were euthanized on the 30th postoperative day, and the other half on the 90th postoperative day to harvest the stoma region. The healing conditions of the stoma after the 2 anastomotic approaches were observed with naked eyes, under light microscope and scanning electron microscope.</p><p><b>RESULTS</b>The stoma leakage rate (50% versus 0% on the 30th postoperative day, 37.5% versus 12.5% on the 90th postoperative day, both P<0.05) and stenosis degree (13.9%±0.3% versus 7.1%±0.3% on the 30th postoperative day, 17.2%±0.4% versus 9.4%±0.4% on the 90th postoperative day, both P<0.01) were significantly higher in group B than in group A. Compared with traditional manual anastomoses, the histological analysis under light and electron microscope showed a more continuous stoma with more regular epithelium proliferation and collagen arrangement, less inflammation in group A.</p><p><b>CONCLUSIONS</b>Magnetic anastomosis stent ensures better healing of the stoma even under the circumstance of severe inflammation.</p>

Exploring the compensation mechanism for public hospitals of county-level during the ongoing health reform / 中华医院管理杂志

The drug zero-profit reform in place has changed the compensation channels for China's public hospitals at the county level.In view of this situation,the paper probed into the present landscape of the compensation mechanism at such hospitals.The authors analyzed five compensation channels,i.e.,the special government budget subsidy,adjustment of medical service charges and rates,health insurance payment,social financing,and income/expenditure management.The study aimed at providing policy proposals and references for sustainable reform at such hospitals.

Optimal microsurgical treatment of dorsum sellae meningioma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>There are few reports of microsurgical treatment of dorsum sellae meningiomas-which, because of location, size, and differences in growth direction, clinical presentations, degree of surgical difficulty, have varied posttreatment sequelae. In pursuit of an optimal microsurgical treatment option for dorsum sellae meningioma patients, we performed a retrospective analysis of eight microsurgery-treated patients in our set up.</p><p><b>METHODS</b>Clinical data of eight microsurgery-treated dorsum sellae meningioma patients were analyzed. Dorsum sellae meningiomas were classified into 2 types based on tumor location, size, and direction of growth. Type I tumors (dorsum sellae-inferior third ventricle type, four cases) were resected by craniotomy via the frontotemporal or orbitozygomatic approach. Type II tumors (dorsum sellae-third ventricle type, 4 cases) were resected by frontal craniotomy via the transcallosal-interforniceal approach.</p><p><b>RESULTS</b>Complete tumor resection was achieved in all the eight patients. In Type I tumor patients, the only postoperative complication was oculomotor nerve palsy. In Type II tumor patients, the postoperative complications included hyperthermia, electrolyte imbalances, endocrinologic disturbances, and hydrocephalus. The mean follow-up was 2.1 years. Four patients returned to normal life and found a job, two were able to live independently, one required assistance, while one died.</p><p><b>CONCLUSIONS</b>Dorsum sellae meningioma surgery is challenging, and resection of Type II tumors is more difficult than Type I tumors. The selection of a suitable microsurgical approach based on tumor type, and the active treatment of postoperative complications are important means of increasing therapeutic efficacy.</p>

Protection of hepatocyte growth factor against hydrogen peroxide-induced mitochondria-mediated apoptosis in rat cortical neurons / 生理学报

Hepatocyte growth factor (HGF) pretreatment could protect multiple cell types from apoptosis induced by various damages including oxidative stress. The present study was designed to investigate the protective effect of HGF on rat cortical neurons against apoptosis induced by hydrogen peroxide (H2O2) in culture, and then to explore whether HGF could influence the mitochondrial pathway of apoptosis. Primary rat cortical neurons were isolated from Sprague-Dawley rats and cultured in serum free medium containing 2% B27 and Neurobasal-A. To mimic the oxidative stress damage, cortical neurons were exposed to 100 mumol/L H2O2 for 4 h. To explore the effects of HGF on the neurons subjected to H2O2 injury, cells were pretreated with HGF 15, 30, 60 ng/mL for 24 h, respectively, and then exposed to 100 mumol/L H2O2 for 4 h. The cell viability was measured by MTT colorimetric assay and cell injury was evaluated by lactate dehydrogenase (LDH) leakage rate. Apoptotic cells were detected by Hoechst 33258 staining and Annexin V-FITC/PI double labeled flow cytometry. The caspase-3 activity was assessed by colorimetry. The alteration of transmembrane potential of mitochondria was determined by confocal laser scanning microscopy. The expression of cytochrome C protein was measured by Western blot analysis. The results showed that H2O2 treatment significantly decreased the cell viability, increased LDH leakage rate and the percentage of apoptotic cells. Pretreatment of HGF at different concentrations (15-60 ng/mL) could remarkably increase the cell viability of neurons. Compared with that of H2O2 group (53.4%+/-7.4%), the cell viabilities of neurons treated with 15, 30, and 60 ng/mL HGF significantly increased to (69.3+/-6.4)%, (77.5+/-6.1)% and (82.9+/-9.3)% (P<0.05), respectively. HGF preincubation also evidently decreased the LDH leakage rate in cortical neurons damaged by H2O2. The results of Hoechst staining revealed that HGF pretreatment could significantly reduce the apoptotic rate of neurons. The apoptotic rate of H2O2 group was (62.8+/-7.1)%, while that of HGF groups decreased significantly to (34.8+/-8.4)%, (23.5+/-3.2)% and (18.6+/-4.5)% (P<0.05), respectively. The data from caspase-3 activity assay indicated that HGF preconditioning could also remarkably decrease the caspase-3 activity of neurons. In addition, in the presence of various concentrations of HGF, the decrease of transmembrane potential of mitochondria in neurons caused by H2O2 injury could be reversed. Moreover, as detected by Western blot analysis, HGF downregulated the expression of cytochrome C protein in neurons. These results suggest that HGF has a protective effect on rat cortical neurons against apoptosis induced by H2O2, which might be related to the inhibition of the mitochondrial apoptotic pathway and the suppression of the caspase-3 activity.

THE CHANGES OF MYOCARDIAL ULTRASTRUCTURE AND CELL MEMBRANE CHANNEL IN UREMIC RATS / 解放军医学杂志

The aim of this study was to explore the myocardial ultrastructure of experimental uremic rats and the effect of myocardiocyte succinate dehydrogenase(SDH) and free cytosolic calcium changes on membrane channel. By making a uremia model in rats, changes of SDH activation was observed by quantitative enzyme cytochemistry methods and membrane calcium channel was checked by laser scanning confocal microscopy. The results showed that with the uremia becoming heavier, myocardiocyte membrane was rolled and broken, myofibrils were swollen and broken. The quantity of mitochondria with SDH products significantly decreased, while activation of cell membrane calcium channel markedly increased and cytosolic calcium piled up. It is suggested that impaired myocardial membrane, decreased contents of mitochondrial function enzyme and cytosolic calcium overload are the pathophisiological basis leading to cardiac dysfunction in uremia.