Objective To identify rodent residues using DNA barcoding technology in plague areas of Shaanxi and to analyze the feasibility of DNA barcoding method.Methods DNA extraction,PCR,electrophoresis and sequence determination and alignment were used to determine the cytochrome C oxidase Ⅰ (COI) gene sequence from 62 residues of 7 species identified by morphology in 8 different parts.COI gene sequence was analyzed using BLAST software of American National Center for Biotechnology Information (NCBI) for sequence homology comparison and the phylogenetic tree was constructed by using the neighbor joining(NJ)method of COI gene sequence.Results In addition to the hair,CO I genes of the feet,the tail,the fur,the muscle,the ribs,the ear,and the eye were amplified,respectively,and the size of amplified fragment was similar to the size of the product with an expected fragment(700 bp),and the fragment was a single band.COI genes of 59 residue specimen were obtained by DNA sequencing and there were high degree of homologies between CO I gene sequences of Meriones unguiculatus,Cricetulus barabensis,Meriones meridianus,Dipus sagitta,Phodopus roborovskii,Rattus norvegicus and Allactaga sibirica and their corresponding host genes in NCBI(99.0％,98.1％,99.8％,98.9％,99.5％,99.1％,98.3％).NJ method showed that 7 NJ phylogenetic trees were constructed with COI sequence of 59 species residues.The CO I sequences of same rodent with different residues were clustered into one group.Condusion DNA barcoding technology can identify host animal residues in plague areas,and the identification results are reliable.

Objective To apply DNA barcoding technology for exploring its taxonomic status and differences in the molecular biology of Cricetulus barabensis in Shaanxi Province.Methods Sixty-five samples of Cricetulus barabensis were collected from Dingbian,Jingbian Counties in northern of Shaanxi and Dali County in Guanzhong plain (Dingbian 58 samples,Jingbian 2 samples,and Dali 5 samples).According to the mitochondrial cytochrome C oxidase subunit I gene (CO I) sequence,the genetic distance was calculated and Neighbor-Joining tree was constructed.Results The genetic distance between two samples (13.16,13.21) and other 56 samples of Dingbian was 9.2％-10.0％.The genetic distance between the 56 samples of Dingbian and Jingbian was less than 1％ and Dali was 7.2％-8.3％;the average intraspecific genetic distance of Jingbian and Dali was less than 1％.The Neighbor-Joining tree showed that all the Cricetulus barabensis samples from the three counties were separated into two large branches.The samples of 13.16,13.21 from Dingbian together were classified into a class and the rest of the samples into another separate branch.At the same time,other samples from Dingbian except 13.16,13.21 and Jingbian were distributed in a small branch,and Dali samples were occupied another small branch.Conclusion Using the DNA barcoding technology,we have determined three subspecies of Cricetulus barabensis in Shaanxi Province,Dingbian has two kinds and Dali has a different subspecies.

We studied the genetic characteristics of Spermophilus in Shaanxi Province,China.The COI,Cyt-b gene were sequenced and the results were compared with those of dauricus from Inner Mongolia Keyouzhong Banner and Zhengxiangbai Banner,and S.alaschanicus from Haiyuan County of Ningxia.And genetic distance was analyzed and Neighbor-Joining tree was built.Results showed that the genetic distance of COI gene sequences between Spermophilus from Dingbian County in Shaanxi and S.alaschanicus in Ningxia was ≤0.5％,and the genetic distance was ranged from 7.9％ to 9.3％ with Citellus dauricus from Inner Mongolia.The genetic distance of Cyt-b gene between Spermophilus from Dingbian County in Shaanxi and S.alaschanicus in Ningxia was ≤2.2％,and ranged from 8.9％ to 11.2％ with Citellus dauricus from Inner Mongolia.The Neighbor-Joining tree of COI,Cyt-b gene showed two major clusters.One of them were clustered by Spermophilus from Dingbian County in Shaanxi and S.alaschanicus in Ningxia,and another one was Citellus dauricus from Inner Mongolia.The Neighbor-Joining tree of COI gene showed that all samples from Shaanxi Province clustered in a group.In conclusion,the Spermophilus in Shaanxi Province were S.alaschanicus.

OBJECTIVETo investigate the varieties of entophytes in different parts of Pinellia ternata.

METHODThe solidified plates were applied for the isolation of the endophytes, and three methods were used for the identification of endophytic fungi.

RESULTEighty four strains of the entophytes were isolated from the P. ternata collected from 3 habitations. Endophytic fungi were morphologically identified belonging to 15 genera, 4 families.

CONCLUSIONIt indicated that the entophytes in P. ternata were diversity and rich, and there were some differences at quantity and species in different organs of P. ternata.

Biodiversity ; Fungi ; classification ; isolation & purification ; physiology ; Phylogeny ; Pinellia ; microbiology ; physiology ; Symbiosis

Objective:To investigate the application value of TRIANGLE operation in radical resection of pancreatic cancer.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 30 patients with pancreatic cancer who underwent TRIANGLE operation in the First Affiliated Hospital of Nanjing Medical University from March 2020 to July 2020 were collected. There were 18 females and 12 males, aged from 41 to 79 years, with a median age of 65 years. After assessment of the resectability and dissection of the hepatoduodenal ligament, the superior mesenteric artery, celiac axis, common hepatic artery, portal vein and superior mesenteric vein of patients were exposed. According to the location of tumor, patients were performed pancreaticoduodenectomy, distal pancreatectomy or total pancreatectomy combined with dissection of putatively tumor-infiltrated lymphatic and neural tissue from the triangular space. During the TRIANGLE operation, it required to at least finish the skeletonization of right semi-circumference of the superior mesenteric artery and celiac axis in cases of pancreatico-duodenectomy, whereas the left semi-circumference in cases of distal pancreatectomy. In principle, both of the superior mesenteric artery and celiac axis were circumferentially skeletonized for cases of total pancreatectomy. Lymphatic, neural and fibrous tissues between the superior mesenteric artery and the celiac artery were completely dissected in all patients. Observation indicators: (1) surgical conditions and postoperative histopathological examination; (2) postoperative recovery situations; (3) follow-up. Regular follow-up was conducted by telephone interview or outpatient examinations to understand tumor recurrence and metastasis of patients up to January 2021. Measurement data with skewed distribution were represented as M (range). Count data were described as absolute numbers or percentages, Fisher exact probability method was used for comparison between groups. Results:(1) Surgical conditions and postoperative histopathological examination: all patients underwent open operation, including 21 cases of pancreaticoduodenec-tomy, 6 cases of distal pancreatectomy, 2 cases of total pancreatectomy, 1 case of mid-segment preserving pancreatectomy. There were 16 cases combined with portal vein-superior mesenteric vein resection and 3 cases combined with left adrenal resection. For the dissection area of superior mesenteric artery-celiac axis, one cycle was conducted in 5 of 30 patients, one and a quarter cycle in 8 patients, one and a half cycle in 8 patients, one and three quarters?two cycles in 9 patients. There were 16 cases with left gastric vein dissection and 14 cases with left gastric vein preservation. The operative time of 30 patients was 287 minutes (range, 165?495 minutes) and the volume of intraoperative blood loss was 275 mL(range, 50?800 mL). Nine patients received intraoperative transfusion of red blood cells or frozen plasma. Postoperative histopathological examination showed that the tumor diameter was 3.4 cm (range, 1.2?7.3 cm), the number of harvested lymph nodes was 20 (range, 9?35), the number of positive lymph nodes was 2 (0?19). Of the 30 patients,20 cases had moderately differentiated tumor and 10 cases had poorly differentiated tumor. R 0 resection was achieved in 9 patients, 1 mm R 1 resection in 17 patients, and R 1 resection in 4 patients. Postoperative pathological T stages: 3 of 30 patents were in stage T1, 18 cases were in stage T2, 5 cases were in stage T3, and 4 cases were in stage T4. Postoperative pathological N stages: 9 of 30 patents were in stage N0, 13 cases were in stage N1, and 8 cases were in stage N2. Postoperative pathological TNM stages: 2 of 30 patents were in stage Ⅰa, 2 cases were in stage Ⅰb, 3 case were in stage Ⅱa, 11 cases were in stage Ⅱb, 12 cases were in stage Ⅲ. (2) Postoperative recovery situations: 20 of 30 patients had postoperative complications, including 6 cases of Clavien-Dindo grade I complications, 9 cases of grade Ⅱ complications, 3 cases of grade Ⅲa complications, 2 cases of grade V complications. The incidence of postoperative diarrhea was 0 in cases with one cycle dissection of tissues around superior mesenteric artery-celiac axis, 1/8 in cases with one and a quarter cycle dissection, 4/8 in cases with one and a half cycle dissection, 9/9 in cases in with one and three quarters-two cycles dissection, showing a significant difference between the four groups ( P<0.05). The incidence of delayed gastric emptying was 5/16 in patients with left gastric vein dissection, versus 1/14 in patients with left gastric vein preservation, showing no significant difference between the two groups ( P>0.05). Nineteen of 30 patients received adjuvant chemotherapy.Twenty-eight of 30 patients were successfully discharged from hospital, with the duration of postoperative hospital stay of 15 days(range, 8?68 days) and the rest of 2 patients died. Three patients had unplanned readmission within postoperative 90 days. (3) Follow-up: all the 28 patients discharged from hospital were followed up for 1.0?9.0 months, with the median follow-up time of 6.5 months. During the follow-up, one patient with locally advanced pancreatic cancer had local recurrence, 9 patients had liver metastasis (including 4 cases with resectable pancreatic cancer, 4 cases with borderline resectable pancreatic cancer, 1 case with borderline resectable pancreatic cancer), and 1 patient with borderline resectable pancreatic cancer had peritoneal metastasis. The rest of 17 patients survived without tumor recurrence. Conclusions:TRIANGLE operation in the radical resection of pancreatic cancer has high radicality and low postoperative local recurrence, but a slightly high morbidity and mortality rate. Its long-term efficacy needs to be further evaluated. It is recommend that this procedure should be performed for selected patients after neoadjuvant chemotherapy in high-volume pancreatic surgery centers.

Objective To investigate the characteristics and risk factors of pulmonary hypertension (PH) in patients with chronic obstructive pulmonary disease (COPD) in plateau areas. Methods To select 562 COPD patients in Qinghai Plateau from January 2017 to December 2020 in the Qinghai Province Cardiovascular and Cerebrovascular Disease Specialist Hospital. The patients were divided according to pulmonary artery systolic pressure (PASP) pulmonary hypertension (PH) group (PASP≥40mmHg) and non-pulmonary hypertension (NPH) group (PASP<40mmHg). The basic characteristics of the two groups of patients were compared, and multivariate logistic regression analysis was used to explore the related factors affecting pulmonary hypertension in COPD patients. Results Among the 562 COPD patients, the total incidence of PH was 29.89% (168/562). Among them, the incidence of PH in patients with grade I COPD was 16.16%, which was significantly lower than that of COPD patients with grade II (34.40%) and grades III to IV (43.86%), and the difference was statistically significant (χ²=30.848, P＜0.001). The percentage of forced expiratory volume in one second (FEV1%), blood oxygen saturation, B-type natriuretic peptide, mean platelet volume (MPV) in the PH group were different from those of NPH group (P<0.05). Multivariate logistic regression analysis showed that FEV1% [OR=1.082 (95%CI: 1.038-1.256)], MPV [OR=1.696 (95%CI: 1.273-2.257)] were risk factors for COPD patients with PH in high altitude areas (P<0.05). Conclusion The incidence of PH is higher in COPD patients in plateau areas, and COPD patients with lower FEV1% and higher MPV are more likely to develop PH.

Objective To apply the DNA barcoding technology for identification on host animal and to establish the host animal DNA bar code database on natural foci of plague in Shaanxi.Methods 139 host animals belonging to 3 orders,6 families and 12 genera and 62 residues belonging to 7 species from 8 different parts of the province,were detected.DNA barcoding technology was used to analyze the DNA CO Ⅰ gene sequence on the natural foci of plague in Dingbian county.Results The intra-specific genetic distance was less than 2％ while the inter-specific distance ranged from 8.9％ to 15.1％.Fourteen major clusters were apparently showed on a Neighbor-Joining tree.Residue samples could be detected regarding the objective gene.Alashan ground squirrel was previously noticed to carry 14 major clusters,which were previously mistakenly named as Citellus dauricus in Dingbian county.Conclusion DNA barcoding technology could overcome the shortcomings caused by the morphological identification so could be used to identify the host animal and residues in the natural focus of plague.

Objective:To investigate the Correlation between ADC combined with serum C-reactive protein (CRP) and delayed encephalopathy after carbon monoxide poisoning (DEACMP), It provides scientific basis for early prediction of DEACMP.Methods:According to the design principle of case-control study, the data of acute carbon monoxide poisoning (ACOP) patients admitted to Shandong Provincial Hospital from December 2017 to December 2021 were retrospectively selected. Among them, patients with DEACMP were selected as the case group, without DEACMP were used as the control group. Univariate and multivariate analyses were performed on the two groups. Receiver operating characteristic curve (ROC) was used to evaluate the diagnostic efficacy of ADC combined with CRP as a combined predictor for disease.Results:A total of 89 patients with ACOP were included, including 33 patients with DEACMP and 56 patients without DEACMP. There were no significant differences in gender, age, smoking, drinking, and underlying diseases (hypertension, coronary heart disease) between groups ( P>0.05). Logistic regression analysis showed that white blood cell count (WBC) ( OR=1.64, 95% CI: 1.19-2.26, P=0.003), CRP ( OR=1.22, 95% CI: 1.03-1.45, P=0.019) and ADC value of central semiovale white matter ( OR=0.99, 95% CI: 0.98-1.00, P=0.010) were associated with DEACMP in patients with ACOP. The ROC curve results showed that the area under the ROC of ADC combined with CRP in the center of semiovale was 0.765 (95% CI: 0.656-0.845), the specificity was 87.9%, the sensitivity was 23.2%, and the cut-off value was 3.5°. Conclusions:WBC, CRP and ADC value of central semiovale are independent factors for DEACMP. ADC value of central semiovale combined with CRP has more clinical value in the early diagnosis of DEACMP. For ACOP patients with DEACMP triggering factors, the diagnosis and treatment awareness of early screening of brain magnetic resonance imaging should be strengthened to avoid DEACMP.

Objective:To evaluate the value of left-sided uncinate process first approach in pancreaticoduodenectomy.Methods:The clinical data of 152 patients who underwent the left-sided uncinate process first approach during pancreaticoduodenectomy at Pancreas Center, the First Affiliated Hospital of Nanjing Medical University from January 2020 to December 2020 were analyzed retrospectively. There were 64 females and 88 males,with age( M( Q R)) of 62.0(14.7)years(range:16.0 to 84.0 years). The clinical date of 117 patients who underwent pancreaticoduodenectomy without using left-sided uncinate process first approach in the same period was selected as the control group,including 65 females and 52 males,with age of 64.0(13.0) years(range:13.0 to 84.0 years). Fisher exact probability method and t test were used to compare the data between the two groups,rank sum test was used for comparison of continuous variables between the two groups. Results:Pancreaticoduodenectomy was successfully performed in 152 patients in left-sided uncinate process first approach group. The operation time was 222.5(77.0) minutes(range:117.0 to 480.0 minutes),the time of uncinate process resection from left-side(the time from jejunum dissection to complete dissociation of the uncinate process) was 11.0(4.5) minutes(range:7.5 to 20.0 minutes),the time of pancreatic head resection (the time from jejunum dissection to pancreaticoduodenal specimen removal) was 26.0(8.5) minutes(range:20.0 to 41.0 minutes),the intraoperative blood loss was 200(150) ml(range:50 to 800 ml),and the intraoperative blood transfusion rate was 9.2% (14/152). Postoperative conditions:The postoperative hospital stay was 12 (9) d(range:6 to 55 d),the overall incidence of postoperative complications was 59.9%(91/152),and there was no perioperative death. Pathological results:The R0 resection rate of periampullary malignant tumor was 64.3%(77/112),with negative rate of uncinate process margin was 91.1%(102/112). The R0 resection rate of pancreatic ductal adenocarcinoma was 46.9%,with negative rate of uncinate process margin was 89.1%(57/64). Compared with the non-left-sided uncinate process first approach group(222.5(77.0) minutes, 9.2%(14/152)),the left-sided uncinate process first approach group had shorter operation time(246.0(94.0) minutes) ( Z=3.964, P<0.01),less intraoperative blood loss (18.8%(22/117))( Z=4.843, P<0.01),and lower intraoperative blood transfusion rate(χ2=5.248, P=0.029). However,there were no significant differences between two groups in postoperative hospital stay( Z=1.682, P=0.093),postoperative overall complications( P=0.549),R0 resection rate of periampullary malignant tumor(χ2=2.012, P=0.156),and negative rate of uncinate process margin(χ2=2.108, P=0.147). Conclusions:The "left-sided uncinate process first approach" could completely resect uncinate process under a direct vision,especially when the uncinate process was behind the superior mesenteric artery or beyond the left lateral margin of the superior mesenteric artery. The "left-sided uncinate process first approach" might increase the negative rate of uncinate process margin and R0 resection rate for periampullary malignant tumor.

Objective:To investigate the clinical efficacy of distal pancreatectomy with celiac axis resection(DP-CAR).Methods:A total of 89 consecutive patients (50 males and 39 females) who were diagnosed with pancreatic body cancer and underwent DP-CAR in Pancreas Center,First Affiliated Hospital of Nanjing Medical University between September 2013 and June 2022 were retrospectively reviewed. There were 50 males and 39 females,with age( M(IQR)) of 63(12) years(range:43 to 81 years). Perioperative parameters,pathology results and follow-up data of these patients were analyzed, χ2 or Fisher′s test for categorical data while the Wilcoxon test for quantitative data. Survival results were estimated by the Kaplan-Meier survival method. Results:Among 89 cases,cases combined with portal vein-superior mesenteric vein or organ resection accounted for 22.5% (20/89) and 42.7% (38/89),respectively. The operative time,blood loss and postoperative hospital stay were 270 (110) minutes,300 (300) ml and 13 (10) days,respectively. The overall morbidity rate was 67.4% (60/89) while the major morbidity was 11.2% (10/89). The increase rate in transient liver enzymes was 42.7% (38/89),3.4% (3/89) for liver failure,53.9% (48/89) for clinically relevant postoperative pancreatic fistula,1.1% (1/89) for bile leak,3.4% (3/89) for chylous leak of grade B and C,11.2% (10/89) for abdominal infection,9.0% (8/89) for postoperative hemorrhage of grade B and C,4.5% (4/89) for delayed gastric emptying,6.7% (6/89) for deep vein thrombosis,3.4% (3/89) for reoperation,4.5% (4/89)for hospital mortality,7.9% (7/89) for 90-day mortality. The pathological type was pancreatic cancer for all 89 cases and pancreatic ductal adenocarcinoma made up 92.1% (82/89). The tumor size was 4.8(2.0) cm, ranging from 1.5 to 12.0 cm. The number of lymph nodes harvested was 14 (13)(range:2 to 33),with a positive lymph node rate of 13.0% (24.0%). The resection R0 rate was 30.0% (24/80) and the R1 (<1 mm) rate was 58.8% (47/80). The median overall survival time was 21.3 months (95% CI: 15.6 to 24.3) and the median disease-free survival time was 19.1 months (95% CI: 11.7 to 25.1). The overall survival at 1-year and 2-year were 69.60% and 39.52%. The median survival time of 58 patients with adjuvant chemotherapy was 24.3 months (95% CI: 17.8 to 32.3) while that of 13 patients without any kind of adjuvant therapy was 8.4 months (95% CI: 7.3 to 22.3). Seven patients accepted neoadjuvant chemotherapy and there was no significant morbidity among them,with a resection rate of R0 of 5/7. Conclusion:DP-CAR is safe and feasible for selective cases,which could be more valuable in improving long-term survival when combined with (neo) adjuvant therapy.