OBJECTIVE To establish the fingerprint of Gerbera delavayi and the methods for the content determination of 11 components in G. delavayi. METHODS High-performance liquid chromatography（HPLC）was adopted to establish the fingerprints of 13 batches of G. delavayi（No. S1-S13）， and the similarities were evaluated according to Similarity Evaluation System of Chromatographic Fingerprint of TCM （2012 edition）， while the common peaks were identified. Hierarchical clustering analysis （HCA）， principal component analysis （PCA） and orthogonal partial least square-discriminant analysis （OPLS-DA） were carried out by using SPSS 25.0 software and SIMCA 14.1 software. The contents of neochlorogenic acid， chlorogenic acid， cryptochlorogenic acid， 3，8-dihydroxy-4-methoxy-2-oxo-2H-1-benzopyran-5-carboxylic acid， caffeic acid， 3-hydroxy-4-methoxy-2- oxo-2H-1-benzopyran- 5-carboxylic acid， luteolin-7-O-β-D-glucoside， isochlorogenic acid A， apigenin-7-O-β-D-glucoside， isochlorogenic acid C and xanthotoxin were determined by HPLC. RESULTS The similarities in HPLC fingerprint of 13 batches of G. delavayi were 0.801-0.994； a total of 38 common peaks were identified and 13 common peaks were identified. The results of HCA showed that S1-S5 and S7 were clustered into one group， S6 into one category， S8 into one category， S9 and S11 into one category， S10， S12 and S13 into one category， and the results of PCA were consistent with them. The results of OPLS-DA showed that variable importance values for the projection of peak 7 （chlorogenic acid）， peak 21 （isochlorogenic acid A）， peak 26 （xanthotoxin）， peak 19 （isochlorogenic acid B）， peak 33， peak 13， peak 23 （isochlorogenic acid C）， peak 2 （new chlorogenic acid）， peak 17 （luteolin-7-O-β-D- glucoside） were greater than 1. The above 11 components had good linearity in their respective detection concentration ranges （r was greater than 0.999）. RSDs of precision， repeatability， and stability tests were not more than 2% （n＝6）. The average recovery rates were 92.54%-105.55%， and the RSDs were 0.83%-1.93% （n＝6）. The average contents of 11 components were 0.744， 5.014， 0.646， 0.431， 0.069， 0.582， 0.979， 2.754， 0.157， 1.284 and 2.943 mg/g， respectively. CONCLUSIONS The constructed HPLC fingerprint and content determination methods are simple， accurate and stable， which can provide reference for quality control of G. delavayi. Xanthotoxin， chlorogenic acid， isochlorogenic acid A， luteolin-7-O- β -D-glucoside， isochlorogenic acid C and new chlorogenic acid can be used as markers for G. delavayi.

Objective:To investigate the impact of age,various hormonal levels,and biochemical markers on penile cavernous body vascular function in patients with erectile dysfunction(ED).Me-thods:A retrospective analysis of clinical data from male patients with ED who underwent color duplex Doppler ultrasonography(CDDU)and intracavernosal injection test(ICI)at the Reproductive Medicine Center of Peking University Third Hospital from January 2020 to August 2023.Data were managed and processed using SPSS 29.0,and a multivariable Logistic regression analysis was conducted.Results:A total of 700 ED patients were included,with 380 showing negative ICI results and 320 positive.In the study,84 patients had a peak systolic velocity(PSV)＜25 cm/s,while 616 had PSV ≥ 25 cm/s;202 patients had end-diastolic velocity(EDV)＞5 cm/s,and 498 had EDV ≤5 cm/s.264 patients had ab-normal PSV and/or EDV results,and 436 had normal results for both.Patients with vascular ED had sig-nificantly lower estrogen levels(t=-3.546,P＜0.001),lower testosterone levels(t=-2.089,P=0.037),and a higher rate of hyperglycemia(x2=12.772,P=0.002)compared with those with non-vascular ED.The patients with arterial ED were older(t=3.953,P＜0.001),had a higher rate of hyperglycemia(x2=9.518,P=0.009),and a higher estrogen/testosterone ratio(t=2.330,P=0.020)compared with those with non-arterial ED.The patients with mixed arteriovenous ED had higher age(t=3.567,P＜0.001),lower testosterone levels(t=-2.288,P=0.022),a higher rate of hyperglycemia(x2=12.877,P=0.002),and a larger estrogen/testosterone ratio(t=2.096,P=0.037)compared with those with normal findings.Multifactorial Logistic regression analysis indicated that higher levels of estrogen were a protective factor for vascular ED(OR=1.009,95％CI:1.004-1.014),and glucose 7.0 mmol/L was a risk factor(OR=0.381,95％CI:0.219-0.661).Older age was a risk factor for arte-rial ED(OR=0.960,95％CI:0.938-0.982).Additionally,older age(OR=0.976,95％CI:0.958-0.993)and glucose levels of 5.6-6.9 mmol/L(OR=0.591,95％CI:0.399-0.876)were also risk fac-tors for mixed arterio-venous ED.Conclusion:Hyperglycemia and aging may impair penile cavernous body vascular function,while higher levels of estrogen may have a protective effect on it.

Objective To explore the value of ultrafast pulse wave velocity(UFPWV)technique for evaluating changes of carotid artery elasticity in Hashimoto thyroiditis(HT)patients with euthyroidism.Methods Conventional ultrasound and UFPWV for carotid artery were prospectively performed in 91 HT patients with euthyroidism(HT group)and 81 healthy subjects(control group).Clinical data and ultrasonic parameters were compared between groups.Spearman correlation analysis was performed to observe the correlations of carotid pulse wave velocity in end of systole(PWV-ES)with clinical indexes and other ultrasonic parameters in HT group.Multiple stepwise linear regression analysis was used to screen the independent impact factors of increased carotid PWV-ES in HT group.Results Significant differences of thyroid peroxidase antibody(TPOAb),thyroglobulin antibody(TgAb),total cholesterol(TC),low density lipoprotein and neutrophil-lymphocyte ratio(NLR)were found between groups(all P＜0.05).The carotid PWV-ES in HT group was significantly higher than that in control group(P＜0.05),while no significant difference of carotid artery intima-media thickness(CIMT)nor carotid pulse wave velocity in beginning of systole(PWV-BS)was found between groups(both P＞0.05).Carotid PWV-ES in HT group was positively correlated with patients'age,body mass index,TPOAb,TC,triglyceride,NLR and CIMT(r=0.217-0.707,all P＜0.05).Patients'age,TPOAb and NLR were all independent impact factors of increased carotid PWV-ES in HT patients with euthyroidism(all P＜0.05).Conclusion UFPWV technique could be used to evaluate changes of carotid artery elasticity in HT patients with euthyroidism,among which PWV-ES was relatively sensitive.

Objective:To study the relationship of pathogenic bacteria in midstream urine culture and stone composition of patients characteristics with infection stones.Methods:Between January 2016 and December 2020, 989 patients with infectious stones who attended Tsinghua Chang Gung Hospital, affiliated with Tsinghua University, for surgical treatment were enrolled in the study. There were 545 male and 444 female patients, with the mean age (48±14) years. The left and right side stones were 396 and 333, respectively. There were 260 bilateral stones, 264 single stones, 334 multiple stones, and 391 deer-stalker-shaped stones. The maximum diameter of stones was (33.4±26.5)mm, combined with diabetes in 109 cases and hypertension in 235 cases. Clean middle-urine was collected for bacterial culture, and intraoperative stone specimens were collected by percutaneous nephrolithotomy (PCNL). Personal characteristics of the patient such as gender, age, body mass index, clinical information such as stone size, location, comorbidities, results of urine culture and stone composition were recorded. The differences of infectious stone composition was analyzed between urease-producing, non-urease-producing bacteria.Results:Among the 989 patients with infectious stones, 259 were pure infectious stones, 131 were mixed infectious stones, and 599 were combined with infectious stone components. Urine cultures were positive in 627(63.4%) patients with infectious stones. The predominant urease-producing bacteria included Ureaplasma urealyticum(94 case), Proteus mirabilis(58 case), and Staphylococcus spp.(36 case). Pure infectious stones were common in Proteus mirabilis, while combined with infectious stone components were common in Ureaplasma urealyticum and Staphylococcus spp. The predominant non-urease-producing bacteria included Escherichia coli(175 case), Enterococcus spp.(76 case) and Streptococcus spp.(35 case). Escherichia coli commonly contained in infectious stone components and pure infectious stones, whereas Enterococcus spp. and Streptococcus spp. commonly contained in infectious stone components. Escherichia coli (61 case), Proteus mirabilis (44 case) and Enterococcus spp.(20 case) were the most common bacteria in 259 cases of pure infectious stones. Escherichia coli (36 case), Enterococcus spp. (14 case) and Ureaplasma urealyticum (10 case) were the most common bacteria in 131 cases of mixed infectious stones. The most common bacteria in 599 cases of combined infectious stones were Escherichia coli (78 case), Ureaplasma urealyticum (68 case) and Enterococcus spp. (42 case).Conclusions:Urease producing bacteria were not common in infectious stones. It was common for the Ureaplasma urealyticum in combined infectious stone components, while Escherichia coli was common in pure and combined infectious stone components.

Objective:To compare the consistency of tomographic infrared spectrum analysis with conventional infrared spectrum analysis for the composition analysis of large-volume of urinary stones in vitro.Methods:Postoperative urinary stone specimens collected from 105 patients admitted to Beijing Tsinghua Changgung Hospital from January 2019 to June 2021 were analyzed, including 81 (77.14%) kidney stones, 16 (15.24%) ureteral stones, and 8 (7.62%) bladder stones. All stones measured ≥0.8 cm in maximum diameter on preoperative imaging. Eighty-four specimens, which were mainly stone fragments, were collected from percutaneous nephrolithotomy and ureteroscopic lithotripsy. These 84 specimens were analyzed and retested for stone composition using conventional infrared spectrum analysis by random multiple sampling. Other 21 renal stone specimens were obtained by laparoscopic lithotomy or standard percutaneous nephrolithotomy after November 1, 2020. These 21 specimens had a maximum diameter of ≥0.8 cm measured postoperatively. Based on intraoperative observation, stone specimens with typical layered structures were chosed. Then, all 21 samples were analyzed and retested by conventional infrared spectrum analysis and tomographic infrared spectrum analysis, respectively. When using tomographic infrared spectrum analysis, we need to take two maximum cross sections with a vertical spacing of these sections >2 mm, then perform multiple points sampling according to the morphological stratification of the first section. If the section's structure was homogeneous, we equidistantly took 2 to 3 samples from the center to the periphery. Otherwise, every layer needed to take a stone sample according to the stratification. Putting all the results of one section together, we obtained complete tomographic infrared spectrum analysis data. Take another coaxial cross-section of the same specimen for retesting. We recorded the characteristics of the three-dimensional distribution of stone composition in 21 stone specimens. Meanwhile, we compared the consistency of the results of conventional infrared spectrum analysis and tomographic infrared spectrum analysis for the same sample.Results:The consistency rate of the conventional infrared spectrum analysis was 56.19% (59/105), and that of tomographic infrared spectrum analysis was 80.95% (17/21). The difference in consistency between two methods was statistically significant ( χ2=4.447, P=0.035). Among 21 specimens, the consistency rate of conventional infrared spectrum analysis was 38.10% (8/21), which was significantly lower than that of tomographic infrared spectrum analysis ( χ2=7.814, P=0.005). Regarding the characteristics of the three-dimensional distribution of the components, the color and crystal morphology of five common types of stone components were different, and layered structure in the cross-section of the stones were observed. When the calculi were of the same composition, they were displayed in different morphology. We observed a trending change in the composition ratio between sublayers from the center to the edge in some compound-composition stones. Conclusions:For the composition analysis of larger-volume urinary stones, tomographic infrared spectrum analysis showed a higher consistency of retesting than conventional infrared spectrum analysis, and the three-dimensional distribution of stone composition had some characteristic features.

Domestic and foreign literatures related to the persistence of SARS-CoV-2 and the re-positive cases infected with SARS-CoV-2 were reviewed, and the characteristics and infectivity of the re-positive cases were analyzed to provide scientific evidence for the improvement of case management and the development of measures to stop the spread of SARS-CoV-2. Existing studies have shown that re-positive rate of SARS-CoV-2 ranged from 2.4% to 19.8%, the median of interval between re-positive detection and discharge was 4-15 days. Following the second course of the disease, the anti-SARS-CoV-2 IgM, IgG and IgA positive rates of the cases were 11.11%-86.08%, 52.00%-100.00% and 61.54%-100.00% respectively, the total antibody and neutralizing antibody positive rates were 98.72% and 88.46%. The viral load of the re-positive cases was lower than that in the initial infection. At least 3 380 re-positive cases have been reported globally. SARS-CoV-2 strains were isolated from the samples of 3 re-positive cases (1 immunodeficiency case and 2 cases with abnormal pulmonary imaging). There were close contacts that were infected by an asymptomatic case taking immunosuppressive agents. In conclusion, the infectivity of re-positive cases infected with SARS-CoV-2 is generally very low. Rare re-positive cases infected with SARS-CoV-2 might cause further transmission. The management approach for the re-positive cases can be based on the assessment of the individual transmission risk according to the pathogen detection results.
Antibodies, Neutralizing ; Antibodies, Viral ; COVID-19 ; Humans ; Immunoglobulin M ; SARS-CoV-2

Objective:To establish virus culture method and full genome sequencing method for severe acute respiratory syndrome coronavirus (2019-nCoV), and to illuminate the variation of 2019-nCoV.Methods:The pharyngeal swab specimens were inoculated into Vero-E6 cells for isolation, and the cytopathic effect were observed day by day, and the result of virus isolation were confirmed by Real-Time RT-PCR. Some isolated viruses were diluted 10 times to detect the virus titer. Full-genome of 2019-nCoV was sequenced with the whole genome capture technology and next generation sequencing technology. Phylogenetic analysis and molecular characterization for obtained 2019-nCoV sequences were undertaken.Results:Twenty-two 2019-nCoV strains were isolated with Vero-E6 cell line in the Biosafety Level-3 Laboratory (BSL-3). Eighteen full-genome sequences obtained (almost 29 000 nucleotide) were analyzed with 99.94%~100% identity and 28 nucleotides and 20 amino acids variations. Phylogenetic analysis showed that the 2019-nCoV sequences belonged to different clades, including clade S and clade L with Europe lineage L. I and Europe lineage L. II.1. Furthermore, 5 of 18 2019-nCoV harbor the D614G mutation.Conclusions:2019-nCoV were successfully isolated with Vero-E6 cells, and partially illuminated the phylogenetic characteristics, which provided a basis for subsequent drug screening and biological characteristics research. Continuous monitoring and analysis of the sequences of new cases would be vital to understand the genetic evolution and rates of substitution of the 2019-nCoV.

Objective The application of nano-carbon tracer technique in radical antegrade modular pancreatosplenectomy (RAMPS) to treat malignant tumors of pancreatic body and tail.To investigate whether radical anterograde modulization of pancreatic body and tail combined with nano-carbon tracer technique is more beneficial to the prognosis of patients.Methods From January 2014 to September 2018,a retrospective analysis was performed in 48 patients in Linyi Central Hospital,all patients underwent RAMPS,all of whom were pathologically proved to be pancreatic carcinoma of the body and tail.Based on whether the nano-carbon lymph node tracer was performed during the operation,the patients were divided into two groups:nano-carbon group (nano-carbon lymph node tracing) and conventional operation group (no nano-carbon lymph node tracer).The differences between the two groups were analyzed,including operative time,intraoperative blood loss,postoperative complications (pancreatic fistula),postoperative hospitalization time,postoperative pathological lymph node status (number of dissection,number of positive lymph nodes,the positive rate of lymph node black staining in nano-carbon group) and survival time after operation.The measurement data were expressed as mean ± standard deviation (Mean ± SD),t-test were used for comparison among groups;Chi-square test was used for comparison of counting data,and Kaplan-Meier test was used for comparison of survival rate.Results There was no significant difference in the time of operation [(4.45 ± 0.59) h vs (4.41 ±0.65) h],the amount of blood lost during operation [(447.39 ± 109.51) ml vs (434.8 ± 76.38) ml] and the time of hospitalization after operation [(16.52 ± 4.12) d vs (16.56 ±2.92) d] between the two groups (P＞0.05).There were 1 case of pancreatic fistula in the nano-carbon group and 2 cases of pancreatic fistula in the convertional operation group,but the difference was not statistically significant (P =0.55).The number of lymph nodes dissected and positive lymph nodes in nano-carbon group were significantly higher than those in conventional operation group [(18.26 ± 2.49) vs (12.16 ± 2.19);(5.52 ± 1.50) vs (3.32 ± 0.85).The difference was statistically significant (P ＜ 0.05).The positive rate of lymph nodes in black stained lymph nodes (38.10％,96/252) was significantly higher than that in non-stained lymph nodes (18.45％,31/168),and the difference was statistically significant (P ＜ 0.05).The average survival time in the nano-carbon group [(25.48 ± 11.74) months] was significantly longer than that in the conventional operation group [(15.80 ± 11.64) months],and the difference was statistically significant (P ＜ 0.05).Conclusions It is safe,effective and feasible to treat malignant tumors of pancreatic body and tail by RAMPS combined with nano-carbon tracer technique.The positive rate of lymph node dissection and the detection rate of total lymph nodes are increased,and the prognosis of patients is improved,to improve the survival time of patients after operation is improved.

Objective To investigate the risk factors of hemorrhagic transformation(HT) in patients with acute cerebral infarction(ACI).Methods A total of 335 patients with ACI from June 2013 to September 2016 was enrolled in this study,including 47 patients in hemorrhagic transform group(HT group) and 288 patients in non-hemorrhagic transformation group(NHT group).The general clinical situation,laboratory indexes,imaging features and treatment measures of the two groups were collected and compared,and then the risk factors of HT in the patients with ACI were analyzed by Logistic regression analysis.Results Univariate analysisshowed that there were significant differences between the two groups in the history of diabetes mellitus,hìstory of atrial fibrillation,NHISS score,systolic blood pressure at admission,fasting blood glucose,glycosylated hemoglobin,low density lipoprotein cholesterol,fibrinogen,infarct location,large area infarction and thrombolytic therapy(P＜0.05).Logistic regression analysis showed that history of atrial fibrillation(OR =2.703,95 ％ CI 1.169-6.250),high fasting blood glucose(OR =2.098,95 ％ CI 1.532-2.875),large area infarction(OR=9.999,95％CI 4.648-21.510) and thrombolytic therapy(OR=6.557,95％CI 1.954-22.003) were independent risk factors for HT.Conclusion The history of atrial fibrillation,high fasting blood glucose,large area infarction and thrombolytic therapy are the risk factors for HT in patients with ACI.Corresponding nursing measures should be arranged to facilitate the disease treatment.

Objective To investigate the protective effects of Ulinastatin on intestinal barrier damaged after cardiopulmonary resuscitation (CPR) in rats in order to illustrate the possible mechanism.Methods Twenty-one male SD rats were divided into three groups randomly (random number) including control group (sham group, n =7), cardiopulmonary resuscitation group (CPR group, n =7) and ulinastatin group (UTI group, n =7).The rats were anesthetized with pentobarbital sodium (45-60 mg/kg) by intraperitoneal injection.The rats of sham group were only treated with endotracheal intubation.Ulinastatin (100 000 U/kg) were injected via caudal vein 2 hours prior to CPR, and cardiac arrest was made in rats and cardiopulmonary resuscitation was carried out in the UTI group, while equivalent volume of sterile saline was used instead in the CPR group.Blood and ileum samples were obtained at 48 hour after restoration of spontaneous circulation (ROSC).The levels of tumor necrosis factor alpha (TNF-α) and interleukin-1β (IL-1β) were assayed by ELISA (enzyme-linked immunosorbent assay), the protein levels of caspase-3 were determined by western blot, the intestinal mucosa were stained by terminaldeoxynucleoitidyl transferase mediated nick end labeling (TUNEL) and ileac mucosa were observed under transmission electron microscope.Data were processed with SPSS 17.0 software.Results The plasma levels of TNF-α and IL-1β were dramatically higher in CPR group than those in other two groups (CPR vs.sham, P ＜ 0.01;CPR vs.UTI, P ＜ 0.05).Moreover, the tight junctions between cells obviously broadened and loosened in the CPR group were found under electron microscope, however, this phenomenon was not obvious in the UTI group.A large number of apoptotic cells were observed by TUNEL assay in the CPR group, but a small number of apoptotic cells were observed in the UTI group.The protein levels of caspase-3 in the UTI group were higher than those in sham group, but lower than those in CPR group (both P ＜ 0.05).Conclusions Ulinastatin has protective effects on the intestinal barrier damaged after cardiopulmonary resuscitation in rats by decreasing the proinflammatory mediators in the blood, reducing the expression of caspase-3and then reducing the numbers of apoptotic intestinal cells.