Aim To investigate the relationship between the brain targeting effect and P-glycoprotein(P-gp)expression level of Danshensu borneol ester(DBE)and the combination use of sodium Danshensu and borneol(SDSS-B).Methods The liquid chromatography mass spectrometry(LC-MS)method was applied to investigate the accumulation of Danshensu(DSS)in rat brain tissues after intravenous injection of DBE,SDSS-B and SDSS.Also their effect on regulating the expression level of P-gp in rat hippocampus was investigated using Western blot.Results The brain targeting effect of DBE,SDSS-B was qualitatively analyzed through the brain distribution of DSS,and the result was DBE(SDSS-B)>SDSS(P<0.01).Meanwhile,the brain distribution of DBE group was slightly lower than SDSS-B group at 15 min,while at 30 min DBE was higher than that of SDSS-B.DBE demonstrated a better slow release property compared to SDSS-B.Western blot analysis indicated that DBE,SDSS-B were more effective in inhibiting the expression of P-gp than SDSS in rat hippocampus(P<0.01 vs control or SDSS group),and the lowest P-gp expression was obtained with(47.58±2.28)％and(46.54±1.41)％at 45 min after administration of DBE or SDSS-B.Once the administration time was extended to 60 min,the inhibitory effect on P-gp expression of DBE was stronger than SDSS-B[(85.04±1.42)％vs(95.29±0.98)％].However,no significant inhibition of P-gp expression in rat hippocampus was found throughout the treatment of SDSS(P>0.05 at 5,15,45,60 min,vs control group).Conclusion An attenuated expression level of P-gp can be realized by DBE and SDSS-B,which is advantageous to their brain targeting.

Objective To explore the effect of Baihe Zhimu Tang(百合知母汤,BZT) on the airway inflammation in bronchial asthma rats.Methods Forty Wistar rats were randomized into control group,asthma model group,BZT group and dexamethasone (DX) group with 10 rats in each group.The bronchial asthma rat model was built by intraperitoneal injection of 10％ ovalbumin (OVA) and ultrasonic atomizing inhalation of 2％ OVA except for the control group.Thirty minutes before every ultrasonic atomizing inhalation,the BZT group was given BZT 3.68 g/(kg · d) by gastric perfusion;the DX group was administered DX injection 1.2 mg/(kg · d) by gastric perfusion;as for the control group and asthma model group,they were given normal saline of the same volume;all groups were intervened for 14 days.After that,HE staining was used to observe the pathological changes of lung tissue,the levels of interleukin-2 (IL-2) and interleukin-4 (IL-4) were measured,and the cell cycle and calcineurin (CaN) activity were observed in separated peripheral blood lymphocytes.Results For the asthma model group,there was a lot of inflammatory cell infiltration in the bronchial wall under light microscope,the content of IL-4 in the plasma was higher than that in the control group,while the content of IL-2 decreased,the proportion of G0/G1 phase lymphocytes decreased,the proportion of S phase and G2/M increased,and the CaN activity of lymphocytes increased significantly (P ＜ 0.05).For the BZT group,there was only a small amount of inflammatory cell infiltration in the lung tissue under light microscope,the IL-4 level in the plasma was lower than that in the asthma model group,while the IL-2 content increased,the proportion of G0/G1 phase lymphocytes increased,the proportion of S phase and G2/M decreased significantly,and the CaN activity of lymphocytes decreased significantly (P ＜ 0.05).Conclusion BZT could reduce lymphocyte proliferation,decrease IL-4 level and reduce airway inflammation by reducing CaN activity in peripheral blood lymphocytes of bronchial asthma rats.

Objective To study the influence of pregnant rats' prenatal chronic stress (PS) on learning and memory of their offspring rats and its possible molecular mechanisms.Methods Pregnant females were individually restrained for 45 min 3 times a day during pregnancy from day 14 to day 21.Control pregnant females were left undisturbed in their home cages.The rat offsprings were randomly assigned to PS group or control group.Males and females were kept for the study separately.The learning and memory of the developing rat offspring in the Morris water maze were examined.The basal levels of corticosterone (COR) and adreno-cortico-tropic-hormone (ACTH) were analyzed by using radioimmunoassay.The Golgi-Cox impregnation technique was used to compare density and morphology of the CA1 hippocampal dendritic spines.Results The escape latency (EL) to find the platform in the control group was significantly less than that in the PS group in female rat offspring (F =4.533,P ＜ 0.05),and the difference was statistically significant on the 5th day (t =2.788,P ＜ 0.01).EL to find the platform in the control group was significantly less than that in the PS group in male rat offspring (F =6.101,P ＜0.05),and the difference was statistically significant on the second day (t =3.051,P ＜ 0.01).In the space exploration experiments of the water maze,the retention time observed for the control group and the PS group in the goal quadrant was similar(P ＞ 0.05).The basal levels of the serum COR in the PS group were higher than those in the control group of female rat offspring(t =3.658,P ＜ 0.01) and the basal levels of the serum ACTH in the PS group were higher than those in the control group of male rat offsprings(t =2.319,P ＜ 0.05).A simplified pattern was observed in the CA1 hippocampal dendritic spines in the PS group,showing a less extent of dendritic arborization and the density was significantly lower than that in the control group(t =-3.072,P ＜ 0.01).Conclusions Altered function of the hypothalamic-pituitary-adrenal axis in the offspring mediates the cognitive alterations observed following prenatal stress should to be associated with the lower density and simplified pattern of CA1 dendritic spines.

Objective To investigate the safety and feasibility of totally laparoscopic transabdominalhiatal approach in the treatment of Siewert type Ⅱ adenocarcinoma of esophagogastric junction (AEG).Methods The retrospective and descriptive study was conducted.The clinicopathological data of 11 patients with Siewert type Ⅱ AEG who were admitted to Affiliated Hangzhou First People's Hospital of Zhejiang University School of Medicine from May 2017 to July 2018 were collected.There were 8 males and 3 females,aged 56-72 years,with an average age of 63 years.Patients underwent radical resection of AEG by totally laparoscopic transabdominalhiatal approach.Observation indicators:(1) surgical situations and postoperative recovery;(2) postoperative pathological examination;(3) follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed to detect postoperative adjuvant chemotherapy,complications,food intake,anastomosis patency,tumor recurrence and metastasis,and survival up to December 2018.Measurement data with normal distribution were presented as Mean±SD,measurement data with skewed distribution were presented as M (range),and count data were represented as absolute number or percentage.Results (1) Surgical situations and postoperative recovery:all the patients underwent totally laparoscopic radical resection of Siewert type Ⅱ AEG by transabdominal-hiatal approach,without conversion to open surgery or perioperative death.Of the 11 patients,8 underwent total gastrectony including 3 combined with splenic hilar lymph node dissection and 3 underwent proximal gastrectomy with double-tract reconstruction.Operation time,time of superior overlap esophagojejunostomy,volume of intraoperative blood loss,time for initial out-of-bed activities,time to first flatus,time to initial liquid diet intake,time of drainage tube removal were respectively (245± 39)minutes,(60± 12) minutes,(75±23) mL,(24± 8) hours,(2.4± 0.5) days,(3.5 ± 0.8) days,(8.2 ± 1.3) days respectively.There was no serious complication including postoperative hemorrhage,anatomotic fistula or death.Three patients had left pleural effusion,and were cured after thoracic drainage.The duration of postoperative hospital stay was (11.0±3.0) days.(2) Postoperative pathological examination:all the 11 patients had negative upper surgical margin.The length of proximal margin,tumor diameter,total number of lymph lodes harvested,and number of lower mediastinal lymph lodes harvested were (2.1 ±0.2) cm,(2.6±0.9) cm,(36.0±4.0)/case and (2.3± 0.8)/case.Pathological examination showed adenocarcinoma in all the 11 patients.pTNM staging:2 cases were in stage Ⅰ B,4 cases in stage Ⅱ A,3 cases in stage Ⅱ B and 2 cases in stage Ⅲ A.(3) Follow-up and survival situations:11 patients were follow-up for 6-19 months,with a median time of 9 months.Chemotherapy regimeus were formulated according to the pathological examination.Nine patients received postoperative adjuvant chemotherapy,and 2 in stage Ⅱ B received no postoperative adjuvant chemotherapy.During the follow-up,11 patients had no obvious reflux symptom or choking feeling,and the anastomosis was patent as evaluated by oral contrast agent and gastroscopy.There was no tumor recurrence and metastasis or death in the 11 patients.Conclusion Totally laparoscopic transabdominal-hiatal approach applied in the radical resection is safe and feasible for the treatment of Siewert type Ⅱ AEG,with good short-term outcomes.

Objective:Based on 108, 591 cases of pediatric emergency visits in a Level Ⅲ Grade A women and children′s hospital in Guangzhou area, we analyze the disease spectrum and epidemiological characteristics, and summarize the characteristics of patient flow changes.These investigations will provide an basis for scientific decision-making for manpower and material resource management of pediatric emergency and hospital workflow design.Methods:The children admitted to the Pediatric Emergency Department of the Zhujiang New Town District of Guangzhou Women and Children′s Medical Center from October 2016 to September 2018, including night emergency and inpatient observations, were analyzed according to the admission date, admission time, gender, age, initial diagnosis and etc.Results:There were more boys than girls in the emergency department, whose ratio was 1.46∶1 (64 480∶44 111 cases). The age of children ranged from 0 to 17 years old, with a median of 11 (23, 48) months.The age distribution was mainly under 5 years old, accounting for 84.14% (91 336/108 591). During the whole year, the number of children in July was the most, accounting for 10.53% (11 433/108 591), and the children in February were the least, accounting for 6.04% (6 555/108 591). The highest visit time of the whole day was 22-23 pm, accounting for 18.83% (20 443/108 591). The most of the diagnosis was respiratory disease, accounting for 53.83% (66 522/123 576). A total of 1 057 critically ill children were received, accounting for 0.97% (1 057/108 591). A total of 911 accidental injuries were received, accounting for 0.84% (911/108 591). Acute upper respiratory tract infection was the most among all diagnoses, accounting for 34.47% (42 541/123 576).Conclusion:Children in the pediatric emergency department of Guangzhou Women and Children′s Medical Center are mainly under 5 years old, and the number of children is the most in July of the year.The main disease is respiratory diseases.Medical staff can be trained according to the actual situation, and the disease spectrum can be updated in time to provide convenience for emergency rescue and improve service level.

Objective:To investigate the application value of totally laparoscopic trans-abdominal-hiatal approach in the radical resection of Siewert type Ⅱ adenocarcinoma of esophago-gastric junction (AEG).Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 45 patients with Siewert type Ⅱ AEG who were admitted to the Affiliated Hangzhou First People′s Hospital, Zhejiang University School of Medicine from May 2017 to December 2020 were collected. There were 28 males and 17 females, aged from 35 to 85 years, with a median age of 64 years. All patients underwent radical resection of AEG by totally laparoscopic trans-abdominal-hiatal approach with gastrointestinal anastomosis using proximal gastrectomy with double-tract anastomosis or total gastrectomy with esophagojejunointestinal anastomosis and digestive reconstruction using transdiaphragmatic-hiatal superior overlap esophagojejunostomy. Observation indicators: (1) surgical and postoperative situations; (2) postoperative histopathological examination; (3) follow-up and survival situations. Follow-up using outpatient examination and telephone interview was performed to detect survival of patients and tumor recurrence and metastasis up to March 2021. Measurement data with normal distribution were presented as Mean± SD and measure-ment data with skewed distribution were presented as M(range). Results:(1) Surgical and post-operative situations: all 45 patients underwent radical resection of AEG by totally laparoscopic transabdominal-hiatal approach successfully, including 35 cases undergoing total gastrectomy with esophagojejunointestinal anastomosis and 10 cases undergoing proximal gastrectomy with double-tract anastomosis. The total operation time, time of lower mediastinal lymph node dissection, time of superior overlap esophagojejunostomy, volume of intraoperative blood loss, time for initial out-of-bed activities, time to first flatus, time to initial liquid diet intake, time to drainage tube removal of the 45 patients were (235±32)minutes, (25±8)minutes, (45±10)minutes, (70±13)mL, (20±8)hours, (2.3±0.2)days, (2.6±0.8)days and (6.2±1.1)days, respectively. Eleven of 45 patients under-went postoperative complications and none of patient died during perioperative period. The post-operative duration of hospital stay of 45 patients was (10±3)days. (2) Postoperative histopatho-logical examination: all 45 patients had negative upper surgical margin. The length of proximal margin, tumor diameter, total number of lymph lodes harvested and number of lower mediastinal lymph nodes harvested were (2.5±0.5)cm, (2.9±0.8)cm, 35.0±4.0 and 2.4±0.8, respectively. Patholo-gical examination showed adenocarcinoma in all 45 patients with pTNM staging as 5 cases of stage ⅠB, 8 cases of stage ⅡA, 21 cases of stage ⅡB and 11 cases of stage ⅢA. (3) Follow-up and survival situations: 45 patients were followed up for 3 to 46 months, with a median follow-up time of 26 months. During follow-up, 8 of 45 patients died. Of the 37 patients survived, 3 cases underwent liver metastasis and 3 cases underwent bone metastasis, lung metastasis or peritoneal metastasis respec-tively.Conclusion:Total laparoscopic transabdominal-hiatal approach is safe and feasible in the treatment of Siewert type II AEG with a satisfactory clinical efficacy.

Background and objective Epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs) have be-come ifrst-line treatment drugs for lung adenocarcinoma patients with EGFR gene mutations. Signiifcant interindividual varia-tions in response rate, progression-free survival (PFS), and overall survival (OS) have been observed. hTe expression level of nuclear factor erythroid-2-related factor 2 (Nrf2) is related to chemoresistance against platinum drugs. Nrf2 overexpression can inhibit the sensitivity of EGFR-TKIs in cells with EGFR-sensitive mutations. hTe aim of this study is to investigate the protein expression level of Nrf2 in lung adenocarcinoma patients with EGFR gene mutations and to elucidate the correlation between Nrf2 expression and response rate of ifrst-line EGFR-TKIs, as well as PFS and OS. Methods Immunohistochemical analysis of Nrf2 in tumor specimens was performed on 31 patients with stage III or IV adenocarcinoma harboring EGFR gene mutations. Results hTe Nrf2-positive rate was 77.4%, whereas Nrf2 nuclear high-expression rate was 38.7%. hTe nuclear expression level of Nrf2 was signiifcantly correlated with response rate (RR) and PFS of EGFR-TKIs (P<0.05), but not with gender, age, smok-ing, differentiation, and OS (P>0.05). hTe Nrf2-positive level was signiifcantly correlated with PFS and OS of EGFR-TKIs (P<0.05), but not with gender, age, smoking, differentiation, EGFR gene mutation status, and RR (P>0.05). hTe PFS and OS of patients with Nrf2-positive expression were signiifcantly shorter than those in patients with negative expression (P<0.05). Furthermore, the nuclear expression level of Nrf2 was the independent predictive factor for EGFR-TKI-induced PFS, and the Nrf2-positive level was the independent predictive factor for EGFR-TKI-induced OS (P<0.05). Conclusion hTe expression level of Nrf2 is signiifcantly correlated with response rate (RR) of EGFR-TKIs, PFS, and OS. hTerefore, Nrf2 may be a useful biomarker in predicting response of EGFR-TKIs in patients with advanced-stage lung adenocarcinoma harboring EGFR gene mutations.

A split-plot experiment was conducted to study the optimum planting density of Nuomi yam in directional groove shallow cultivation. Nuomi yam was the main local cultivar of Dioscorea. The study of this paper may help to give valuable instructions of local production. Three different planting densities and shallow groove diameter were designed in the experiment. The results showed that the fresh weight of aerial part per plant, yield and planting benefit of Nuomi yam tuber were the highest when the planting density was 12 000 plants/hm~2 and the diameter of shallow growth groove was 11.0 cm, while they were the lowest when the planting density was 22 500 plants/hm~2 and the diameter of shallow growth groove was 7.5 cm. Along with the increase of planting density, the fresh weight of aerial part, tuber, fresh yield, benefit and commercial tuber rate of Nuomi yam also decreased. The commercial tuber rate of Nuomi yam of 11.0 cm diameter in shallow growing pot was the highest, while it was the lowest with no shallow growing pot. On the contrary, the content of amylopectin and mucin was the highest when there was no shallow groove, and was the lowest when the diameter of shallow groove was 7.5 cm, although there was no significant difference between the two treatments. Therefore, Nuomi yam with the directional cultivation combination of planting density of 12 000 plants/hm~2 and shallow groove diameter of 11.0 cm showed high yield, good appearance quality, high planting benefit and no obvious adverse effect on internal quality.
Dioscorea

Objective:To evaluate the feasibility and efficacy of 68Ga-PSMA PET/CT-guided targeted prostate biopsy for the diagnosis of clinically significant prostate cancer(csPCa). Methods:This retrospective analysis allocated 89 patients with elevated PSA levels between 4.0-20.0 ng/ml to PET group(n=48) or TRUS group(n=41) between September 2017 and June 2019. Patients with PSMA-avid lesions (SUV max≥8.0) underwent PET-TB via a single-puncture percutaneous transgluteal approach (n=19), while patients with negative PSMA-PET underwent systematic TRUS-GB (n=29). Patients in the TRUS group who did not get 68Ga-PSMA PET/CT examination underwent TRUS-GB directly (n=41). The mean age, prostate volume, PSA value of PET group and TURS group were (71.2±9.1) years vs. (68.0±12.0) years, (62.9±29.1)ml vs. (65.4±38.9)ml , 8.8(6.6, 13.6) ng/ml vs. 9.8(7.1, 13.1)ng/ml, respectively (all P>0.05). The diagnostic efficacy and difference of PCa and csPCa between the two groups were compared. PET-TB adopts a new mode of percutaneous gluteus approach and carries out precise image fusion of PSMA-PET/CT and pelvic CT in the same machine and in the same position (prone position). Under the direct guidance of CT, the biopsy is performed with only one precise puncture. Results:PCa and csPCa were detected in 27/89(30.3%)and 20/89(22.5%)in all patients. PET group detected significantly more cases of PCa and csPCa than those of TRUS group [PCa: 41.7%(20/48) vs. 17.1%(7/41), χ2=6.328; csPCa: 33.3%(16/48) vs. 9.8%(4/41), χ2=7.055, P<0.01]. Of 19 patients with PSMA-PET positive, PET-TB detected 16 cases of PCa(84.2%) by a single needle puncture, and the proportions of cancer tissues were ≥80% in 2, 50%-79% in 8, and <50% in 6 cases. Among these, Gleason score was underestimated by biopsy histopathology in 2 patients. Of 3 patients with PET-TB negative, 1 case of low-risk PCa(Gleason 3+ 3) was detected by complementary TRUS-GB. The sensitivity, specificity, positive predictive value, negative predictive value, accuracy of 68Ga-PSMA PET/CT(SUV max≥8.0) for the diagnosis of csPCa were 73.9%(14/19), 93.1%(27/29), 87.5%(14/16), 81.3%(26/32)and 85.4%(41/48), respectively. For PET-TB, only one patient had slight symptoms of haematuria after the puncture, no cases of hematochezia, hemospermia, urinary retention or pelvic infection were observed. Conclusions:68Ga-PSMA PET/CT is a feasible novel puncture technique that may serve as a triage tool for prostate biopsy, and PET-TB may improve the detection rate of csPCa compared with TURS-GB, especially in patients with serum PSA 4.0-20.0 ng/ml.

Single-cell or low-input multi-omics techniques have revolutionized the study of pre-implantation embryo development.However,the single-cell or low-input proteomic research in this field is rela-tively underdeveloped because of the higher threshold of the starting material for mammalian embryo samples and the lack of hypersensitive proteome technology.In this study,a comprehensive solution of ultrasensitive proteome technology(CS-UPT)was developed for single-cell or low-input mouse oocyte/embryo samples.The deep coverage and high-throughput routes significantly reduced the starting material and were selected by investigators based on their demands.Using the deep coverage route,we provided the first large-scale snapshot of the very early stage of mouse maternal-to-zygotic transition,including almost 5,500 protein groups from 20 mouse oocytes or zygotes for each sample.Moreover,significant protein regulatory networks centered on transcription factors and kinases between the MII oocyte and 1-cell embryo provided rich insights into minor zygotic genome activation.