Objective:To analyze the growth characteristics of different genotypes of Japanese encephalitis virus (JEV) in different cell lines, and to provide scientific basis for the selection of cell lines in the study of JEV.Methods:BHK-21, Vero, C6/36, PK-15, DF-1, N2a, SH-sy5y and MDCK cell lines were selected. The proliferation ability of genotype 1 (NX1889 strain), genotype 3 (P3 strain) and genotype 5 (XZ0934 strain) JEV in these cell lines was evaluated by plaque assay and RT-qPCR.Results:Significant cytopathogenic effects (CPE) were observed in BHK-21, Vero, C6/36, DF-1, N2a and PK-15 cell lines across all three JEV genotypes. However, no significant differences in CPE characteristics were observed within the same cell line. SH-sy5y and MDCK cell lines did not show significant CPE, but virus proliferation was detected in SH-sy5y cell line, while MDCK cell line were found to be insensitive to JEV. No significant difference was observed in the proliferation curves of G1, G3 and G5 JEV in BHK-21, Vero and SH-sy5y cell lines. In C6/36 and PK-15 cell lines, the titer of G1 JEV was higher than that of G3 and G5. In DF-1 cell line, G5 demonstrated a higher titer than the other two genotypes, whereas in N2a cell line, G5 showed a lower titer than the other two.Conclusions:There are differences in the proliferation of three different genotypes of JEV in different cell lines, which can provide reference for the study of JEV in different directions.

Through reviewing ancient and modern literature， the textual research of Anemarrhenae Rhizoma（AR） has been conducted to verify the name， origin， changes in production areas， quality evaluation， harvesting and processing methods， so as to provide reference for the development and utilization of the famous classical formulas containing AR. Through the herbal textual research， AR was first published in Shennong Bencaojing， and has been used as the proper name for this herb for generations， and the mainstream source of AR used for generations is the rhizome of Anemarrhena asphodeloides. The high-quality production areas that have been revered throughout the ages are Hebei， Shanxi， Shaanxi， Inner Mongolia and Fangshan district of Beijing， etc. In recent times， AR produced in Yixian county of Hebei province（Xiling Zhimu）， is better known and is regarded as a very good source. At present， cultivated AR is mainly produced in Yixian county and Anguo of Hebei province， Bozhou of Anhui province and other places. The medicinal parts of AR in ancient and modern times are all rhizomes， and the quality is better if it has thick flesh， hard wood， yellow outer color and white section color. The harvesting time recorded in ancient medical books is usually in lunar February and August， with exposure to dryness， while modern harvesting is spring and autumn. The processing methods of the past dynasties were mainly to remove the hair when using， avoid iron when cutting， process with wine or salt water， while the two main specifications in modern times are raw and salted products. Based on the systematic research， it is recommended that the dried rhizome of A. asphodeloides in the famous classical formulas be used for AR. If the original formula specifies processing requirements， it should be operated according to the requirements， if the processing requirements are not indicated， the raw products can be used as medicine.

【Objective】 To investigate the sperm retrieval rate (SRR) of microdissection testicular sperm extraction (M-TESE) in patients with non-obstructive azoospermia (NOA) caused by different causes. 【Methods】 A retrospective analysis was performed on 225 NOA patients during Jan.2020 and Dec.2022. The relation between SRR and patients’ age,body mass index (BMI),testicular volume,endocrine hormones and different etiological classifications were analyzed. 【Results】 According to whether sperm was obtained by surgery,the patients were divided into two groups,including 107 cases in the sperm group and 118 cases in the non-sperm group. There were no significant differences in patients’ age,testicular volume and levels of endocrine hormones between the two groups (P>0.05). According to the different causes,NOA patients with mumps history,cryptorchidism history,AZFc deletion or Klinefelter syndrome (KS) had higher SRR,while idiopathic NOA patients had the lowest SRR (P<0.05). 【Conclusion】 M-TESE is an effective treatment of NOA. There is no correlation between SRR and patients’ age,MBI,testicular volume and levels of endocrine hormones. NOA caused by different etiological classifications may have different SRR.

Objective:In this study, the collected mosquito samples were subjected to viral isolation to identify the species and branch characteristics of arboviruses in five regions of Shanxi Province.Methods:Eight arboviruses in mosquito samples collected from July to September 2020 were detected by real-time fluorescent quantitative PCR, and virus isolation was carried out through cell culture. Virus isolates were identified and analyzed by molecular biology and bioinformatics method.Results:We detected 1 batch of positive samples of Japanese encephalitis virus, 2 batches of positive samples of Culex flavivirus and 8 batches of positive samples of Nam Dinh virus among 121 batches of mosquito samples. Seven virus isolates were isolated, numbered: SX-YJ-Cxp-4、SX-YJ-Ars-2、SX-YJ-Cxp-1、SX-LY-Cxp-10、SX-GP-Ars-5、SX-GP-Cxp-2、SX-GP-Cxp-4, all of which were identified as Nam Dinh virus, and the whole genome sequencing was performed on one of them, and the result showed that Shanxi Nam Dinh virus isolate and Yunnan Nam Dinh virus isolate belonged to the same evolutionary branch.Conclusions:Nam Dinh virus was isolated and identified on the specimen from Shanxi province for the first time.

Objective:To understand the prevalence of human parechovirus (HPeV) in neonates of Hunan Provincial People’s Hospital, and analyze genetic evolutionary characteristics.Methods:From June to September 2021, fecal samples of inpatient neonates were collected in Hunan Provincial People′s Hospital. TaqMan real-time qPCR and RT-PCR were used for HPeV screening and genotyping. High-throughput sequencing and PCR were used to obtain whole genomes. Phylogenetic analysis was performed after sequencing.Results:A total of 123 fecal samples of neonates were collected, of which 22 were HPeV positive, with 17.89% positive rate. All the strains belonged to the HPeV-1 genotype. One full-length genomic sequence of 7 269 bp were obtained, and provisionally named Hunan/HPeV/2021, which has the highest nucleotide identity with known HPeV-1 genotype, with 86.6%-91.9% nucleotide identity. The nucleotide and amino acid identity of open reading frame (ORF) with known similar sequences were 90.3%-92.6% and 97.3%-98.3%, respectively. The phylogenetic analysis showed that Hunan/HPeV/2021 belongs to the HPeV-1 genotype, which is clustered into the same clade as the popular HPeV-1 strains in China.Conclusions:HPeV has a high prevalence in inpatient neonates of Hunan Provincial People’s Hospital and belong to the HPeV-1 genotype.

Objective : To investigate the effect of long non⁃coding RNA (LncRNA) anoctamin 1 antisense RNA⁃1 (ANO1⁃AS1) on the proliferation and apoptosis of esophageal squamous cell carcinoma (ESCC) cells and its possible mechanisms. Methods : Silenced ANO1⁃AS1 lentivirus was transfected in ESCC cells TE⁃1 and EC109. Subsequently, the expression levels of ANO1⁃AS1 and calcium⁃activated chloride channel protein 1 (ANO1) in the cells were detected by qRT⁃PCR. CCK⁃8 and colony formation assays were used to detect the proliferation of TE⁃1 and EC109 cells. ANO1 positively related expressed genes were obtained from the LinkedOmics database and then the gene set was enriched for pathways and possible pathways were validated. The expression levels of proliferating cell nuclear antigen(PCNA), P53 protein, apoptosis⁃related protein ( Bax and Bcl⁃2), ANO1 protein and phosphatidylinositol⁃3⁃kinase/protein kinase B (PI3K/Akt) pathway⁃related protein were assessed by Western blot. Results: After transfection of lentivirus with silent expression function, the expression level of ANO1⁃AS1 was significantly reduced in TE⁃1 and EC109 cells (P < 0. 05); After down⁃regulation of ANO1⁃AS1, compared with the negative control group, the proliferation ability of ESCC cells was reduced (P < 0. 05) and the rate of clone formation decreased (P < 0. 05); Western blot results showed that, compared with negative controls, the expression of PCNA decreased, the expression of oncogene P53 protein increased ( P < 0. 05 ), the expression of proteins ( Bax) increased, Bcl⁃2 decreased and the levels of phosphorylation of the pathway proteins PI3K and Akt decreased (P < 0. 05) . Conclusion Knockdown of ANO1⁃AS1 can decrease proliferation and promote apoptosis in ESCC, which may be achieved by affecting PI3K/Akt pathway activation.

Background::Pre-operative assessment with high-resolution magnetic resonance imaging (MRI) is useful for assessing the risk of local recurrence (LR) and survival in rectal cancer. However, few studies have explored the clinical importance of the morphology of the anterior mesorectum, especially in patients with anterior cancer. Hence, the study aimed to investigate the impact of the morphology of the anterior mesorectum on LR in patients with primary rectal cancer.Methods::A retrospective study was performed on 176 patients who underwent neoadjuvant treatment and curative-intent surgery. Patients were divided into two groups according to the morphology of the anterior mesorectum on sagittal MRI: (1) linear type: the anterior mesorectum was thin and linear; and (2) triangular type: the anterior mesorectum was thick and had a unique triangular shape. Clinicopathological and LR data were compared between patients with linear type anterior mesorectal morphology and patients with triangular type anterior mesorectal morphology.Results::Morphometric analysis showed that 90 (51.1%) patients had linear type anterior mesorectal morphology, while 86 (48.9%) had triangular type anterior mesorectal morphology. Compared to triangular type anterior mesorectal morphology, linear type anterior mesorectal morphology was more common in females and was associated with a higher risk of circumferential resection margin involvement measured by MRI (35.6% [32/90] vs. 16.3% [14/86], P = 0.004) and a higher 5-year LR rate (12.2% vs. 3.5%, P = 0.030). In addition, the combination of linear type anterior mesorectal morphology and anterior tumors was confirmed as an independent risk factor for LR (odds ratio = 4.283, P = 0.014). Conclusions::The classification established in this study was a simple way to describe morphological characteristics of the anterior mesorectum. The combination of linear type anterior mesorectal morphology and anterior tumors was an independent risk factor for LR and may act as a tool to assist with LR risk stratification and treatment selection.

BACKGROUND: Postoperative chylous ascites is an infrequent condition after colorectal surgery and is easily treatable. However, its effect on the long-term oncological prognosis is not well established. This study aimed to investigate the short-term and long-term impact of chylous ascites treated with neoadjuvant therapy followed by rectal cancer surgery and to evaluate the incidence of chylous ascites after different surgical approaches. METHODS: A total of 898 locally advanced rectal cancer patients treated with neoadjuvant chemoradiotherapy followed by surgery between January 2010 and December 2018 were included. The clinicopathological data and outcomes of the patients with chylous ascites were compared with those of the patients without chylous ascites. The primary endpoint was recurrence-free survival (RFS). To balance baseline confounders between groups, propensity score matching (PSM) was performed for each patient with a logistic regression model. RESULTS: Chylous ascites was detected in 3.8% (34/898) of the patients. The incidence of chylous ascites was highest after robotic surgery (6.9%, 6/86), followed by laparoscopic surgery (4.2%, 26/618) and open surgery (1.0%, 2/192, P = 0.021). The patients with chylous ascites had a significantly higher number of lymph nodes harvested (15.6 vs. 12.8, P = 0.009) and a 3-day longer postoperative hospital stay (P = 0.017). The 5-year RFS rate was 64.5% in the chylous ascites group, which was significantly lower than the rate in the no chylous ascites group (79.9%; P = 0.007). The results remained unchanged after PSM was performed. The chylous ascites group showed a nonsignificant trend towards a higher peritoneal metastasis risk (5.9% vs. 1.6%, P = 0.120). Univariate analysis and multivariate analysis confirmed chylous ascites (hazard ratio= 3.038, P < 0.001) as an independent negative prognostic factor for RFS. CONCLUSIONS Considering the higher incidence of chylous ascites after laparoscopic and robotic surgery and its adverse prognosis, we recommend sufficient coagulation of the lymphatic tissue near the vessel origins, especially during minimally invasive surgery.
Chylous Ascites/etiology* ; Humans ; Incidence ; Laparoscopy ; Rectal Neoplasms/surgery* ; Retrospective Studies ; Risk Factors ; Robotic Surgical Procedures/adverse effects*

Objective:To investigate the effectiveness and safety of soothing moisturizing repair cream on acne depressed scar exfoliative fractional laser wound repair.Methods:From October 2018 to June 2020, the Department of Dermatology, Qingdao Haici Hospital Affiliated to Qingdao University took 33 patients with acne depressed scars as the research object, including 8 males and 25 females, aged from 20 to 36 years (29.6±8.6) years. The left and right face comparison method was adopted. After laser operation, the trial side was given a soothing moisturizing repair cream, and the control side was given a placebo. By collecting the patient's facial pictures and objective skin data before and after the laser operation, 1 h, 1 d, 3 d, 7 d, 21 d, and combined with the researcher's semi-subjective evaluation and patient's subjective evaluation the wound skin reaction and wound healing were observed.Results:At 1, 3, 7, 21 days after laser operation, the skin water content of the test side was higher than that of the control side ( P<0.05), and the skin water loss was lower than the control side ( P<0.05); at 3, 7, 21 days, the skin pigment of the test side was lower than the control side ( P<0.05); at 3, 7 d, the test side skin erythema index was lower than the control side ( P<0.05); at 1, 3, 7 d, the test side wound skin erythema, edema, dryness and tightness, etc. were better than the control side ( P<0.05). The duration of pain, crusting time, scab removal time, and complete healing time of the wound on the test side were shorter than those on the control side ( P<0.05). The patient's satisfaction with the moisturization and comfort of the nursing products on the trial side was better than that on the control side ( P<0.05). Conclusions:There is no adverse reaction to the soothing moisturizing repair cream after laser surgery, which can better inhibit skin inflammation, reduce post-inflammatory pigmentation, promote skin healing, and help repair the wound after laser surgery.