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Objective: To analyze the core functional component groups (CFCG) in Yinchenhao Decoction (YCHD) and their possible pathways for treating hepatic fibrosis based on network pharmacology., Methods: PPI data were extracted from DisGeNET, Genecards, CMGRN and PTHGRN to construct a weighted network using Cytoscape 3.9.1. The data of the chemical components in YCHD were obtained from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform (TCMSP), and the potential active components and targets were selected using PreADMET Web server and SwissTargetPrediction. A fusion model was constructed to obtain the functional effect space and evaluate the effective proteins to identify the CFCG followed by GO and KEGG pathway enrichment analyses for all the targets. In cultured human hepatic stellate cells (LX-2 cells), the cytotoxicity of different compounds in YCHD was tested using CCK-8 assay; the effects of these compounds on collagen α1 (Col1a1) mRNA expression and the pathways in 20 ng/mL TGF-β1-stimulated cells were analyzed using RT-qPCR and Western blotting., Results: A total of 1005 pathogenic genes, 226 potential active components and 1529 potential targets in YCHD and 52 potential targets of CFCG were obtained. Benzyl acetate, vanillic acid, clorius, polydatin, lauric acid and ferulic acid were selected for CCK-8 verification, and they all showed minimal cytotoxicity below the concentration of 200 μmol/L. Clorius, polydatin, lauric acid and ferulic acid all effectively inhibited TGF-β1-induced LX-2 cell activation. At the concentration of 200 μmol/L, all these 4 components inhibited PI3K, p-PI3K, AKT, p-AKT, ERK, p-ERK, P38 MAPK and p-P38 MAPK expressions in TGF-β1-induced LX-2 cells., Conclusion: The therapeutic effect of YCHD on hepatic fibrosis is probably mediated by its core functional components including benzyl acetate, vanillic acid, clorius, polydatin, lauric acid and ferulic acid, which inhibit the PI3K-AKT and MAPK pathways in hepatic stellate cells.

Objective: To evaluate the predictive value of combined serum levels of trimethylamine N-oxide (TMAO) and trimethyllysine (TML) for poor prognosis in patients with heart failure. Methods: This single-center prospective cohort study included hospitalized patients with heart failure and complete baseline data from the Department of Cardiology at Ruijin Hospital, Shanghai Jiao Tong University School of Medicine from June 2017 to December 2020. Patients were categorized into four groups based on median serum levels of TMAO and TML after admission: TMAO low level TML low level group (TMAO<9.7 μmol/L, TML<0.73 μmol/L), TMAO low level TML high level group (TMAO<9.7 μmol/L, TML≥0.73 μmol/L), TMAO high level TML low level group (TMAO≥9.7 μmol/L, TML<0.73 μmol/L) and TMAO high level TML high level group (TMAO≥9.7 μmol/L, TML≥0.73 μmol/L). The primary endpoint was a composite endpoint of cardiovascular death and readmission for heart failure. Multiple factor Cox regression analysis was conducted to evaluate the correlation between serum TMAO and TML levels and poor prognosis in patients with heart failure. Results: A total of 471 patients with heart failure were included, with an mean age of (62.5±12.0) years and a median follow-up time of 1.61 (1.06, 2.90) years. Multivariate Cox regression analysis showed that after adjusting for age, gender, and traditional risk factors, the TMAO high level TML high level group had a higher incidence of primary endpoint events compared to the TMAO low level TML low level group ( HR =1.71, 95% CI 1.05-2.77, P =0.03). Conclusion: Elevated serum levels of both TMAO and TML can effectively predict the occurrence of long-term adverse events in patients with heart failure.

Objective: To analyze the efficacy of second-line regimens and prognostic factors in patients with first-relapsed multiple myeloma (MM) treated with bortezomib, cyclophosphamide, and dexamethasone (BCD). Methods: A retrospective cohort study. Clinical data were collected in first-relapsed MM patients after BCD treatment from three tertiary hospitals in north China from July 2009 to October 2022. Patients were classified according to the second-line regimen into the immunotherapy group, single novel agent group [either proteasome inhibitor (PI) or immunomodulatory drug (IMiD)], combination treatment group (both PI+IMiD), and traditional treatment group. Responses to second-line regimens and survival data were analyzed. The Kaplan-Meier method was used for survival analysis and the Cox proportional risk model was used for univariate and multivariate analyses. Results: A total of 217 patients were enrolled including 8.8% (19/217) in the immunotherapy group, 48.4% (105/217) in the PI/IMiD group, 29.9% (65/217) in the PI+IMiD group, and 12.9% (28/217) in the traditional treatment group. The median age was 62 years (range 31-83 years) and 56.2% (122/217) were males. The overall response rates (ORRs) in the four groups were 94.7% (18/19) vs. 56.2% (59/105) vs. 73.8% (48/65) vs. 32.1% (9/28) ( χ 2 =24.55; P <0.001), respectively. The progression-free survival (PFS) of the second-line regimens (2ndPFS) was 17.7 vs. 9.0 vs. 9.2 vs. 4.6 months ( χ 2 =22.74; P <0.001), respectively, among which patients in the PI/IMiD and PI+IMiD groups had comparable 2ndPFS ( χ 2 =1.76; P =0.923). Patients with high-risk cytogenetic abnormalities (HRCAs) achieved the longest 2ndPFS of 22.0 months in the immunotherapy group ( χ 2 =15.03; P =0.002). Multivariate analysis suggested that immunotherapy ( HR =0.11, 95% CI 0.05-0.27), achievement of efficacy of partial response or better ( HR =0.47, 95% CI 0.34-0.66), and non-aggressive relapse ( HR =0.25, 95% CI 0.17-0.37) were independent prognostic factors of 2ndPFS. Conclusion: In this real-world study, immunotherapy was associated with a more favorable efficacy and PFS for first-relapsed MM patients after BCD treatment, with similar outcomes in patients with HRCAs.

Objective: To identify the transmission relationship between HIV infection cases the non-marital non-commercial heterosexual contact in Zhejiang Province. Methods: When HIV positive was informed during January 2020 to January 2022, the staff conducted an epidemiological investigation to collect cases information on sociodemographic characteristics, mobility information, past HIV testing history, high-risk sexual behaviors, sexual partners, and etcetera. At the same time, 6-8 ml of blood from the new diagnosis of people infected with HIV before antiviral treatment was collected to separate the bleeding plasma. pol gene was amplified by nucleic acid extraction and PCR, sequenced by Sequencer 5.0 software, and Cytoscape 3.6.0 software was used to draw HIV molecular transmission network. Results: From January 2020 to January 2022, 88 HIV infected individuals were found in Pujiang County, of which 74 were transmitted through heterosexual transmission, of which 31 were infected through non-marital non-commercial heterosexual contact. Preliminary case studies have found that three female cases have engaged in unprotected non-marital non-commercial heterosexual contact with one male case. Among the 4 infected individuals, 2 of their spouses tested positive for HIV antibodies. Molecular transmission network monitoring was carried out on 65 newly diagnosed cases of heterosexual transmission with acquired sequences, forming 9 transmission clusters. The largest cluster contained 10 cases. A total of 11 HIV-infected individuals were involved in this HIV cluster epidemic. They were 3 males and 8 females, all over 50 years old and were farmers or rural housewives. They were traced to 7 sexual partners (6 negatives of HIV, 1 undetected). Among the 18 respondents' sexual social network relationships, there were 6 couples, 8 permanent partners, and 3 temporary partners. Among 11 HIV infected individuals, there were 9 cases of non-marital non-commercial heterosexual transmission and 2 cases of intramarital transmission. The epidemiological association between 7 non-married non-commercial heterosexual partners and case 2 (56-year-old male farmer), 3 cases confirmed by epidemiological investigation and molecular transmission cluster results, 3 cases confirmed by molecular transmission cluster and epidemiological investigation results, and 1 case confirmed by epidemiological investigation results. Conclusions: The transmission mode of this cluster epidemic was to spread HIV through heterosexual sex with a male case as the core, then cause the transmission within marriage and between fixed sexual partners. The combination of epidemiological investigation and molecular transmission network traceability survey supports the conclusion of this study.

Dracaena marginata is a widely cultivated horticultural plant in the world, which has high ornamental and medicinal value. In this study, the whole genome of leaves from D . marginata was sequenced by Illumina HiSeq 4000 platform. The chloroplast genome were assembled for functional annotation, sequence characteristics and phylogenetic analysis. The results showed that the chloroplast genome of D . marginata composed of four regions with a size of 154 926 bp, which was the smallest chloroplast genome reported for Dracaena species to date. A total of 132 genes were identified, including 86 coding genes, 38 tRNA genes and 8 rRNA genes. Codon bias analysis found that the codon usage bias was weak and there was a bias for using A/U base endings. 46 simple sequence repeat and 54 repeats loci were detected in the chloroplast genome, with the maximum detection rate in the large single copy region and inverted repeat region, respectively. The inverted repeats boundaries of D . marginata and Dracaena were highly conserved, whereas gene location differences occurred. Phylogenetic analysis revealed that D . serrulata and D . cinnabari form a monophyletic clade, which was the closest relationship and conformed to the morphological classification characteristics. The analysis of the chloroplast genome of D . marginata provides important data basis for species identification, genetic diversity and chloroplast genome engineering of Dracaena .

Objective: To study the clinical efficacy of Jujing Decoction (JJD) based on the theory of "yin forming" in the treatment of teratospermia and its effects on semen parameters, sperm morphology, sperm DNA fragmentation index (DFI) and the ultrastructure of sperm mitochondria., Methods: We randomly divided 40 patients with teratospermia into a blank control group (n = 16) and a JJD group (n = 24), the former treated by reproductive health guidance and the latter with JJD, respectively. After 12 weeks of intervention, we observed the clinical effects and changes in the semen volume, sperm concentration, percentages of progressively motile sperm (PMS) and morphologically abnormal sperm (MAS), sperm DFI and the ultrastructure of sperm mitochondria, and analyzed the data obtained based on the "yin forming" theory., Results: The total effectiveness rate was significantly higher in the JJD than in the blank control group (87.50% vs 6.66%, P < 0.05). Compared with the baseline, the patients after treated with JJD showed significant improvement in semen volume (［3.27 ± 1.14］ vs ［4.16 ± 1.84］ ml, P < 0.05), sperm concentration (［38.85 ± 15.88］ vs ［39.35 ± 14.58］ × 10⁶/ml, P < 0.05), PMS (［37.47 ± 2.74］% vs ［42.55 ± 7.07］%, P < 0.05), MAS (［97.43 ± 1.01］% vs ［95.52 ± 0.85］%, P < 0.05), acrosomal integrity (［45.27 ± 5.14］% vs ［51.88 ± 4.48］%, P < 0.05), and DFI (［25.53 ± 6.89］% vs ［17.68 ± 2.38］%, P < 0.05), while the blank controls exhibited only slightly increased semen volume (［3.67 ± 1.77］ vs ［4.40 ± 1.78］ ml) and sperm concentration (［37.24 ± 9.31］ vs ［38.32 ± 10.54］ ×10⁶/ml), but no significant improvement in the other parameters (P> 0.05). Before treatment, folded and discontinuous sperm acrosomal membrane was observed under the transmission electron microscope in the sperm mitochondria of the two groups of patients, the mitochondria stacked and differently sized in the neck and disorderly arranged and some obviously swollen around the tail microtubule. After treated with JJD, the ultrastructure of sperm mitochondria was significantly improved, with relative continuity and integrity of the acrosomal membrane and well-arranged and relatively intact and uniform-sized mitochondria in the neck and tail., Conclusion: Jujing Decoction can increase the semen volume, sperm concentration and percentage of progressively motile sperm, reduce the percentage of morphologically abnormal sperm and sperm DFI, improve sperm acrosomal integrity, maintain the stability of sperm ultrastructure and protect the structure of sperm mitochondria.

Synthetic biology, a course with a sound theoretical system and a wide application range, plays a role in the cultivation of innovative talents in the field of bioengineering. To this end, we have set up a synthetic biology course in our university. First, according to the concept of imparting basic knowledge, highlighting innovative practice, and keeping up with cutting-edge progress, we assembled a high-level teaching team for synthetic biology. The team constantly adjusted and optimized the course contents and achieved a novel and reasonable course system. Second, we introduced frontier cases of synthetic biology reported in high-level journals, as well as breaking news in this field in classroom teaching, which enriched the teaching contents and aroused students' interest. Third, taking these cases as the breakthrough point, we guided students to in-depth discussions through the learning-centered teaching mode to improve students' abilities of critical thinking and theoretical innovation. In summary, the course has achieved good teaching outcomes and improved the cultivation of innovative talents. Therefore, we share our work with peer teachers, aiming to give new insights into the teaching reform of synthetic biology and other related courses.

Percutaneous pulmonary puncture guided by computed tomography (CT) is one of the most effective tools for obtaining lung tissue and diagnosing lung cancer. Path planning is an important procedure to avoid puncture complications and reduce patient pain and puncture mortality. In this work, a path planning method for lung puncture is proposed based on multi-level constraints. A digital model of the chest is firstly established using patient's CT image. A Fibonacci lattice sampling is secondly conducted on an ideal sphere centered on the tumor lesion in order to obtain a set of candidate paths. Finally, by considering clinical puncture guidelines, an optimal path can be obtained by a proposed multi-level constraint strategy, which is combined with oriented bounding box tree (OBBTree) algorithm and Pareto optimization algorithm. Results of simulation experiments demonstrated the effectiveness of the proposed method, which has good performance for avoiding physical and physiological barriers. Hence, the method could be used as an aid for physicians to select the puncture path.

Edible bird's nests (EBNs), a food of animal origin, are temporary nests built by swiftlets to foster their offspring. As EBNs and their products are widely accepted by consumers, the safety of hormones in EBNs has also received increasing attention. The establishment of a method for hormone analysis in EBNs and the investigation of hormone levels based on the analytical method are the most effective measures to eliminate any safety concerns. In this study, a multi-residue method was developed for the simultaneous determination of 45 hormones in EBNs, including estrogens, progesterones, androgens, and cortical hormones. EBN samples (1.0 g) were weighed into 50 mL polypropylene centrifuge tubes and mixed with 8 mL of pure water. Then, the samples were extracted twice with 15 mL of acetonitrile and ethyl acetate (1∶1, v/v) under ultrasonic-assisted conditions for 30 min, and the protein in the EBN samples was precipitated at 4000 r/min for 5 min. The clear supernatants were loaded onto a hydrophilic-lipophilic balanced (HLB) SPE column, which was previously activated with methanol (3 mL) followed by pure water (3 mL). The cartridge was washed with 3 mL of pure water and 3 mL of 50% methanol aqueous solution. The hormones were eluted with 3 mL of methanol. A rapid analysis was performed using high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The hormones in the extracting solution were separated on a Phenomenex Kinetex C 18 column (100 mm×2.1 mm, 2.6 μm) and eluted by gradient elution with acetonitrile-0.1% formic acid aqueous solution (ESI + ) or acetonitrile-water (ESI - ). Qualitative and quantitative analyses were performed using the multiple reaction monitoring (MRM) mode. The HPLC-MS/MS results showed good linearity in the range of 0.20-20.0 μg/L with correlation coefficients ( R 2 ) ≥0.9990. For the 45 hormones, the limits of detection (LODs, S/N ≥3) were 0.04-0.70 μg/kg and the limits of quantification (LOQs, S/N ≥10) were 0.16-2.00 μg/kg. The recoveries of five hormones, namely, fluorometholone, budesonide, aldosterone, fluocinonide, and ethinylestradiol, were 40.2%-63.6%. Owing to the low recoveries, this method might be suitable only for qualitative testing of the five hormones. The recoveries of the other 40 target analytes were between 72.2% and 112.3% at spiked levels of 2.0, 4.0, and 20.0 μg/kg with relative standard deviations (RSDs) of 2.5%-11.6%. The method is characterized by easy operation, rapidness, high precision, and high sensitivity for the 40 compounds. Thus, this method is suitable for determination of the 40 hormones from EBNs for qualitative testing and quantitation. The proposed method was used to analyze 1021 EBN samples from Malaysia, Indonesia, Thailand, and Vietnam from 2017 to 2021. Only three hormones, progesterone, boldenone, and androstenedione, were identified in the EBN samples, while the levels of the other hormones were lower than their individual LODs. The detected rates of progesterone, boldenone, and androstenedione were 100%, 79%, and 89%, respectively. The contents of progesterone, boldenone, and androstenedione in the EBN samples were 0.097-3.58 μg/kg, 0-0.096 μg/kg and 0-1.77 μg/kg, respectively. The levels of hormones in EBNs were compared with those in eggs, pure milk, and dairy products, which were also animal-derived foods. Androstenedione was detected in all egg samples monitored, and its content was higher than that in EBN samples, pure milk, and dairy products. The content of boldenone was similar among the four products investigated in this study. Based on risk assessment using progesterone, the dietary intake was found to be 3.54 μg/d from milk >1.09 μg/d from eggs >0.0030 μg/d from EBNs. The results showed that the levels of hormones in EBNs were much lower than those in eggs, milk, and dairy products for daily consumption. Based on this investigation, the health effect of the hormones in EBNs may be insignificant.

Objective: To observe the clinical efficacy of Jujing Decoction combined with lipoic acid in the treatment of asthenospermia and teratospermia., Methods: Fifty patients with asthenospermia and teratospermia meeting the inclusion criteria were randomly divided into a blank control (n = 10) and a medication group (n = 40), the former provided with fertility guidance and the latter treated with Jujing Decoction combined with lipoic acid. After 12 weeks of treatment, the clinical therapeutic effects were evaluated by comparing the semen volume, sperm concentration, percentages of progressively motile sperm (PMS) and morphologically abnormal sperm (MAS), rate of acrosome integrity, sperm DNA fragment index (DFI) and pregnancy rate between the two groups., Results: Compared with the control group, the medication group achieved a significantly higher overall effectiveness rate (10% ［1/10］ vs 88.89% ［32/36］, P < 0.05) and pregnancy rate (0% ［0/10］ vs 8.33% ［3/36］, P < 0.05) after treatment. The medication group also showed remarkably increased PMS from (21.04 ± 6.49)% to (32.66 ± 7.05)%, decreased MAS from (98.31 ± 1.28)% to (96.52 ± 1.11)%, elevated acrosome integrity from (42.18 ± 16.67)% to (60.42 ± 11.61)%, and reduced sperm DFI from (21.92 ± 6.96)% to (12.37 ± 3.79)%, all with statistically significant differences compared with the blank control group., Conclusion: Jujing Decoction combined with lipoic acid can significantly improve sperm motility, reduce MAS and DFI and increase the pregnancy rate through antioxidant stress, and has a high clinical safety.

Objective: To investigate the relationship between HIV-infected men who have sex with men and their sexual partners in Zhejiang province. Methods: A cross-sectional survey was conducted to recruit newly confirmed HIV/AIDS among MSM from 2015 to 2017, including sexual partner identification and molecular epidemiological study. Plasma was collected to extract RNA, and the pol gene of HIV-1 was amplified by RT-PCR/nested PCR. Phylogenetic tree and molecular transmission cluster were analyzed to identify the transmission relationship between sexual partners. Results: A total of 937 HIV/AIDS among MSM were recruited to promote HIV testing for their sexual partners, and 173 positive sexual partners were identified. 50.8% (61/120) of the gene sequences were clustered among the positive sex partners. Seven pairs of clustered sex partners combined with the results of recent infection preliminarily determined the transmission direction. In the clusters, there were statistical differences between the partners who were diagnosed in the same year ( OR =12.190, 95% CI : 1.563-95.054) or with current residence in the different districts ( OR =17.054, 95% CI : 1.742-166.982). Conclusions: Combined with a molecular transmission network, HIV test for the sexual partners of HIV/AIDS among MSM can improve the accurate tracking of cases and preliminarily determine the direction of transmission, according to the results of recent infection. It is suggested that after HIV is confirmed for HIV/AIDS among MSM, HIV tests should be carried out as soon as possible for their sexual partners, including a cross-regional sexual partner tracking test, which is helpful to improve the tracing procedure.

Objective: To analyze the relationship between infection and transmission of HIV/AIDS, explore the characteristics of the HIV-1 transmission network, and provide evidence for formulating targeted prevention and control measures. Methods: The newly confirmed HIV/AIDS in Jiaxing from 2017 to 2018 were selected for the study. The sociodemographic and infection transmission-related information was gathered. The blood samples were collected and RNA was extracted, RT-PCR and nested PCR amplified the pol gene of HIV-1. The phylogenetic tree was constructed by Mega 6.0 software for subtype analysis. The genetic distance between strains was calculated, and the molecular transmission network was drawn by using Cytoscape 3.6.0 software. The results of the epidemiological study based on the molecular transmission network were analyzed. Results: There were fifteen HIV-1 gene subtypes in 517 cases of HIV/AIDS. The main subtypes were CRF01&#95;AE (37.1%)、CRF07&#95;BC (36.2%) and CRF08&#95;BC (11.8%). Under the 1.0% gene distance threshold, 87 molecular clusters were established, and the total clustering rate was 45.8% (237/517). The cluster risk factors were 60-81 age group (compared with 14-24 age group, OR =2.690, 95% CI :1.058-6.844), married (compared with unmarried, OR =1.698, 95% CI :1.003-2.875), CRF07&#95;BC subtype (compared with CRF01&#95;AE, OR =2.203, 95% CI :1.426-3.404). The largest molecular cluster, CRF07&#95;BC-1, consisted of 50 cases (including 21 cases confirmed in 2017 cases and 29 confirmed in 2018). Multivariate analysis showed that characteristics of unmarried (compared with married, OR =2.482, 95% CI :1.140-5.402), gays (compared with heterosexual, OR =3.163, 95% CI :1.543-6.483), cases with high-risk transmission (compared with other cases, OR =7.631, 95% CI :1.783-32.654), confirmed in Nanhu and Pinghu districts of Jiaxing (compared with other districts, OR =2.225, 95% CI :1.074-4.608) were risks for entering the largest molecular cluster. There were seven suspected high-risk transmission MSM in the largest molecular cluster. The first debut of homosexual behavior was from 2010 to 2018, and the median number of gay partners was 20 ( P 25 , P 75 ：10, 100) within two years before being diagnosed. Six reported a recent history of looking for gay partners in an MSM dating place in Nanhu district. Conclusions: The HIV-1 subtypes of newly confirmed HIV/AIDS were diverse, mainly sporadic in Jiaxing from 2017 to 2018. There were geographical aggregation characteristics and suspected some high-risk transmission cases, suggesting that rapid transmission and targeted intensive intervention are needed.

Objective: To retrospectively analyze the clinical outcomes of single unrelated cord blood transplantation (UCBT) in children with high risk and refractory acute myeloid leukemia (AML) . Methods: Between June 2008 and December 2018, a total of 160 consecutive pediatric patients with AML received single UCBT (excluding acute promyelocytic leukemia) . Myeloablative conditioning (MAC) regimen were applied. All patients received a combination of cyclosporine A (CsA) and mycophenolate mofetil (MMF) for the prophylaxis of graft -versus- host disease (GVHD) . Results: The cumulative incidence of neutrophil cells engraftment at day +42 and platelet recovery at day +120 was 95.0% (95% CI 90.0%-97.5%) at a median of 16 days after transplantation (range, 11-38 days) and 85.5% (95% CI 83.3%-93.4%) with a median time to recovery of 35 days (range, 13-158) , respectively. Incidence of grades Ⅱ-Ⅳ and Ⅲ-Ⅳ acute GVHD and chronic GVHD were 37.3% (95%CI 29.3%-45.2%) , 27.3% (95% CI 20.0%-35.0%) and 22.4% (95% CI 15.5%-28.7%) , respectively. The transplant-related mortality (TRM) at 360 day was 13.1% (95% CI 8.4%-18.9%) . The 5-year cumulative incidence of relapse was 13.8% (95% CI 8.5%-20.3%) . The 5-year disease-free survival (DFS) and overall survival (OS) were 71.7% (95% CI 62.7%-77.8%) and 72.2% (95% CI 64.1%-78.7%) , respectively. The 5-year GVHD and relapse free survival (GRFS) was 56.1% (95% CI 46.1%-64.9%) . The 5-year cumulative recurrence rates of CR1, CR2, and NR groups were 5.3%, 19.9%, and 30.9% ( P =0.001) , and the 5-year OS rates were 79.9% (95% CI 70.3%-86.7%) , 71.1% (95% CI 50.4%-84.4%) and 52.9% (95% CI 33.0%-69.3%) ( χ (2)=7.552, P =0.020) , respectively. Conclusions: For pediatric patients with high risk and refractory AML, UCBT is a safe and effective treatment option, and it is favorable to improve the survival rate in CR1 stage.

Danggui Buxue Decoction is a classic prescription of Qi and blood tonification, which is mainly applied in treating fatigue, internal damage Qi weakness, blood deficiency, and outward going of floating Yang. Modern pharmacology shows that it can promote hematopoiesis, regulate immunity, and protect heart and cerebral vessels. The prescription, often used for the treatment of anemia and other diseases in clinic, is composed of Astragali Radix and Angelicae Sinensis Radix at a dosage ratio of 5∶1. It is a modern compound prescription for invigorating Qi and generating blood. Based on the review of the chemical constituents, pharmacological effects, and clinical applications of Danggui Buxue Decoction, its Q-marker was predicted and analyzed according to the &quot;five principles&quot; of Chinese medicine Q-marker--quality transmissibility and traceability, ingredient specificity, component validity, component measurabi-lity, and formula compatibility environment. The results suggested that calycosin, calycosin-7-O-β-D-glucoside, formononetin, ononin, astragaloside A, ferulic acid, and ligustilide could be used as Q-markers of Danggui Buxue Decoction, which provides reference for establishing the quality system of Danggui Buxue Decoction.

Brazilein is among the main chemical constituents of Caesalpinia sappan . It has diverse pharmacological activities. Modern pharmacological studies have shown that the compound has antitumor, anti-inflammatory, antibacterial, antioxidant, immunomodulatory, and other pharmacological activities. Brazilein is often used as a stain in various industries. The separation of brazilein by traditional column chromatography will not only result in contamination of the chromatographic column materials, but also lead to loss of the active ingredient. Countercurrent chromatography is an advanced liquid-liquid chromatographic separation technique. It has been widely used for natural product separation and isolation as it offers several advantages, such as low solvent consumption, a highly selective solvent system, and high recoveries. Typical countercurrent chromatography techniques include centrifugal partition chromatography (CPC), high-speed countercurrent chromatography (HSCCC), and high performance countercurrent chromatography (HPCCC). It is well known that choosing a suitable solvent system is vital in countercurrent separation. Therefore, two methods were introduced for choosing a suitable solvent system. One is the generally useful estimation of solvent systems (GUESS) method, which employs thin-layer chromatography (TLC) to identify a suitable solvent system with minimal labor for the rapid purification of target compounds, and another is the Shake-Flash method. The solvent system could be determined by observing the distribution of the sample in the upper and lower phases. Two kinds of solvent systems were screened using the TLC-GUESS and Shake-Flash methods, and tested through the analysis mode of the HPCCC instrument. The results showed that chloroform-methanol-water (4:3:2, v/v/v) was the optimal solvent system for HPCCC separation. A total of 15.2 mg of brazilein and 5.7 mg of caesappanin C were obtained from an ethyl acetate extract with high purities (95.6% and 89.0%, analyzed by HPLC) in one step using the preparation mode of HPCCC, the reversed-phase liquid chromatography mode with the apparatus rotated at 1600 r/min, a flow rate of 10 mL/min, separation temperature of 25℃, and detection wavelength of 285 nm. Their structures were determined by spectroscopic and spectrometric analyses. Brazilein stained the solid packing material in the column and was difficult to elute. The results showed that the use of HPCCC for the separation of brazilein can not only prevent the loss of target active ingredients in Caesalpinia sappan , but also shorten the separation and purification times and improve the operating efficiency. Therefore, HPCCC can be used for the separation and preparation of other pigment compounds in Caesalpinia sappan and other dye plants.

Objective: To investigate the characteristics of subtype diversity and transmission on HIV-1 among 12 to 30 years old student MSM in Zhejiang province. Methods: A total of 290 newly diagnosed HIV infected student MSM were selected as the research objects for molecular studies on HIV, in Zhejiang province during 2013 to 2015. Data on epidemiology and plasma samples of these people were collected. HIV-1 nucleotide sequences of pol gene regions were amplified using the RT-PCR/nested PCR method and sequenced. Phylogenetic analysis was performed to determine the HIV-1 genotypes. Characteristics of transmission mode among these cases were also analyzed. Results: A total of 290 cases, 50.3 % were diagnosed in Hangzhou and 81.0 % had college or above degrees. 178 sequences including 10 subtypes, were obtained, with the main subtypes as CRF01&#95;AE (49.4 % , 88/178) and CRF07&#95;BC (39.3 % , 70/178). A total of 18 molecular transmission clusters were formed (42 cases, cluster size from 2 to 4), with the proportions of clusters as 23.6 % (42/178). 61.9 % (26/42) of student MSM with their schools located in the same district within the transmission clusters. Their sexual partners would include both student MSM and non-student MSM. The proportion of clusters among middle school students was 38.2 % (13/34), higher than that of college students (20.1 % , 29/144) ( χ (2)=4.996, P <0.05). Conclusions: The HIV-1 subtypes of student MSM in Zhejiang province appeared diversity, which indicated with the diversity of sources of infection. The geographical distribution of cluster cases is relatively centralized. In order to effectively control the spread of AIDS, more attention should be paid to the sexual partners involved and to specific programs on intervention.

Suanzaoren Decoction is a classic prescription for nourishing the heart and liver, nourishing blood and tranquilizing the mind. It has the functions of sedation and hypnosis, anti-anxiety, anti-depression, anti-convulsion and so on. Modern clinic is mostly used to treat different types of insomnia, depression, neurasthenia, tension headache and vertigo. In this paper, the chemical consti-tuents, pharmacological effects and clinical application of Suanzaoren Decoction are reviewed. Based on this, the quality marker(Q-marker) of Suanzaoren Decoction was predicted and analyzed according to the &quot;five principles&quot; of Q-marker of traditional Chinese medicine--transmission and traceability, specificity, effectiveness, measurability and compatibility environment of compound prescriptions. The results indicated that jujuboside, spinosin, ferulic acid, senkyunolide Ⅰ, sarsasapogenin, mangiferin, liquiritoside and glycyrrhizic acid were predicted and analyzed, and those can be used as Q-markers of Suanzaoren Decoction. Subsequently, the above components can be selected as indicators to control and evaluate the quality of Suanzaoren Decoction and its preparations, and establish a quality traceability system.

Objective: To deliver macro understanding of the latest research progress on clinical trials and approved products of cancer drugs in China in 2019. Methods: The number of clinical trials and related investigational products by domestic and foreign enterprises in 2019 were acquired in the China Food and Drug Administration Registration and Information Disclosure Platform for Drug Clinical Studies, while listed drugs were obtained in the China Food and Drug Administration Query System for Domestic and Imported Drug. Characteristics on stage, scope, indication of those trials, classification and mechanism of involved products, as well as listed anticancer drugs were summarized and depicted. Results: There were 474 cancer drug trials registered in China in 2019, accounting for 21.8% of the total, and 397 (83.8%) were initiated by domestic pharmaceutical enterprises. Overall, international multicenter trials accounted for 13.1%, and phase I trials accounted for 47.3%. Compared with global enterprises, the proportion of international multi-center trials initiated by domestic companies is lower (4.8% vs. 55.8%, P <0.001), and the proportion of phase I clinical trials and bioequivalence trials is higher (51.9% vs. 23.4%, 19.4% vs. 1.3%, P <0.001). An accumulative of 27 cancer types were involved for all the cancer drug trials, and lung cancer, solid tumor, and breast cancer were the most common cancer types, with 103, 95 and 49 trials, respectively. For the three cancer types unique to Chinese population, gastric, liver and esophageal cancer, the total number of initiated trials was 47. For all those trials, there were 335 cancer drug varieties, with 86.0% developed by domestic pharmaceutical enterprises, including 300 therapeutic drugs, 30 adjunctive drugs and 5 preventive drugs. In terms of mechanism, targeted drugs and immune drugs were the most popular, accounting for 74.6% and 20.3%, respectively. In addition, 17 anticancer drugs targeting on 11 cancer types were approved in China in 2019. Conclusions: Clinical trials on cancer drugs in China have ushered a booming era, with large number of innovative agents represented by targeted drugs and immune drugs under clinical development or putting into clinical practice. Those local enterprises are playing more and more critical roles. Strengthening clinical research and development on Chinese unique cancer types is the key direction of future work.

To evaluate and compare of the immunogenicity differences of flagellins FliC and FljB of Salmonella abortus equi, and lay the experimental foundation for the further utilization of the two recombinant proteins, FliC and FljB recombinant proteins were induced, expressed and purified. The purified FliC and FljB were used to immunize mice separately. The antibody level, titer and subtype of mice serum were detected after immunization. Immune-related receptors and histopathological changes were observed in immunized mice after challenged. The recombinant proteins FliC and FljB were successfully induced and expressed. Proteins of about 52 kDa and 42 kDa were purified. High levels of specific IgG antibodies were induced in mice immunized with these two proteins, the antibody level of FljB-immunized group was higher than that of FliC-immunized group, and IgG1 was the dominant subtype of antibody. The challenge protection rate of the FljB-immunized group was 87.5%, higher than that of FliC immunized group. Bacterial loads and observation pathological of FljB-immunized group were better than that of FliC immunized group, the levels of TCR2, TCR4, MHC-I and TCR induced by FljB-immunized group were higher than those of FliC-immunized group. The of immune response induced by FljB group was better than that of FliC group.

Background: The clinical trials of new anti-tumor drugs are prospering in China. The acceptance of clinical trials in patients is an important factor affecting the speed and quality of clinical trials. Previous studies have investigated the acceptance of clinical trials in those cancer patients, who have never participated in a trial. This study is designed to investigate and compare the acceptance and related causes of clinical trials in cancer patients who have once participated in a clinical trial or not., Methods: From June 2018 to April 2019, a standardized questionnaire-based survey was conducted among two groups of cancer patients classified by history of clinical trial participation in Cancer hospital, Chinese Academy of Medical Science, mainly focusing on their overall acceptance of clinical trials and related considerations, including the role of attending doctors, as well as group differences between the two participants., Results: A total of 538 patients were enrolled with an average age of 53.5 years old, 51.1% of whom were males, and 43.3% of whom have never participated in a clinical trial. Overall, 502 patients (93.3%) were willing to or recommend their relatives or friends to participate in clinical trials, and patients with history of clinical trial participation had higher willingness (96.6% vs 90.8%, P=0.008). Patients were most likely to be motivated by expectation of optimal treatment (100.0% vs 99.3%) for both those who had once participated in a clinical trial or those not, respectively followed by financial burden reduction (56.0%) and recommendation by attending doctor (43.7%). The main reasons for unwillingness-to-participate for those who had once participated in a clinical trial were abandoning other treatment options, divided into control group or additional visits, while for those who had never participated in a clinical trial, ineffective treatment or serious adverse reactions were their main concerns. In the decision-making of clinical trial participation, 88% patients highly valued the role of recommendation by attending doctors. Among patients without trial participation history, 60.9% of those had no unwillingness-to-participate expressed that recommendation by attending doctors would change their decisions. The study also reported patients' preferences for information and access to clinical trials., Conclusions: The acceptance of clinical trials in cancer patients in our hospital is generally high, especially in patients who had a history of trial participation. It's of substantial significance to give full play to the role of doctors in improving the acceptance of clinical trials of cancer patients in China.

Background: Early investigation suggested patients' level of awareness regarding clinical trials was related with willingness to participation. This study was intended to evaluate the level of awareness of cancer patients regarding clinical trials and related influencing factors, and to compare the differences of awareness between patients who attended clinical trials before and not., Methods: From Jun, 2018 to April, 2019, standardized question-naires were gathered from cancer patients (attended clinical trials vs not attended clinical trials) in our hospital regarding basic information and 10 other questions about awareness. The level of awareness was evaluated and patients were classified into "low cognition" and "high cognition" groups. Logistic regression analysis was performed to determine whether certain characteristics would predict for awareness., Results: Of the 617 participants, 38.6% have attended clinical trials before. 338 (54.6%) patients had a correct overall understanding of clinical trials, while 44 (7.1%) patients still thought participants were the victim of scientific research. Except for the compensation of medical expenses (51.5% vs 48.7%) and related laws of clinical trials (52.3% vs 45.5%), other parts of understanding were elevated in patients attended clinical trials before comparing with patients who didn't, including significance (86.2% vs 77.6%), risk disclosure (91.2% vs 71.6%), confidentiality (73.2% vs 59.7%), voluntariness (95.8% vs 76.3%), withdrawal (86.6% vs 68.2%) and expenses (62.8% vs 39.2%). The proportion of participants who understand these components did not increase even in 239 patients who had attended clinical trials before. Participants who attended clinical trials before (OR=1.83, 95%CI: 1.11-3.00), unmarried/divorced (OR=5.04, 95%CI: 1.73-14.66), retired (OR=2.53, 95%CI: 1.16-5.50) had a higher level of awareness, while patients who had bad impression with doctors (OR=0.43, 95%CI: 0.26-0.72) had lower awareness., Conclusions: The current level of awareness for clinical trials of cancer patients in our hospital was relatively low, even in patients who had attended clinical trials before. It's necessary to improve patients' awareness of clinical trial by promoting harmony relationship between patients and doctors, as well as by enhancing related propagation. Strengthening the adequacy and efficacy of informed consent in clinical trials also needs to be achieved in the future.

Objective To prepare the monoclonal antibodies (mAb) against hemagglutinin of H4 subtype avian influenza virus (AIV), and develop a sandwich ELISA for the detection of H4 subtype AIV. Methods The BALB/c mice were immunized with inactive H4 subtype AIV. A mAb against H4 subtype AIV, designated as 6G4, was obtained by cell fusion, hemagglutination inhibition (HI) screening and subcloing. Immuofluorescence cytochemistry and Western blotting were used to detect the reactivity of 6G4 with H4 subtype AIV, and the specificity, broad spectrum and stability of 6G4 were characterized by HI assay. Subclass of 6G4 was determined by kit. With chicken polyclonal antibody against H4 subtype AIV as coated antibody, 6G4 mAb as capture antibody and HRP-labeled goat anti-mouse IgG as the enzyme-labeled antibody, a sandwich ELISA for the detection of H4 subtype AIV was established by optimization of the reaction conditions and serial verification. Results 6G4 belonged to IgG1 subclass, and the light chain belonged to κ. It could secrete antibody stably and had good reactivity, specificity, broad spectrum and stability. ELISA based on 6G4 was specific, sensitive, accurate and suitable for the detection of a large number of samples. Conclusion We successfully achieved the anti-H4 subtype AIV mAb, and developed the sandwich ELISA for the detection of H4 subtype AIV.

Objective: Using field epidemiological investigation and molecular analysis to construct the molecular transmission network of human immunodeficiency virus/acquired immunodeficiency syndrome cases (HIV/AIDS) newly diagnosed in Huzhou in 2017, Zhejiang Province. Methods: A total of 160 participants were obtained through a web-based system from Chinese Center for Disease Control and Prevention (CCDC) with the features of diagnosed in Huzhou in 2017 who also had been collected samples for the first follow-up. The basic information of demographic characteristics and risk factors was extracted from the website. RNA was extracted from plasma samples of untreated cases, followed by RT-PCR and nest-PCR for pol gene amplification, sequencing. Phylogenetic tree was constructed by MEGA software for HIV gene subtyping. TN93 model was used for calculating the distance between two sequences. Cytoscape software was used for drawing molecular transmission network. And then an epidemiological survey was conducted to cases in the primary cluster. Results: A total of 138 sequenced individuals (86.3%) were acquired from 160 individuals. Among which, 123 (89.1%) were male. The highest proportion of subtype was CRF07&#95;BC (60, 43.5%), followed by CRF01&#95;AE (46, 33.3%), and with four cases of Unique Recombinant Form (URF, CRF01&#95;AE and CRF07&#95;BC) and one case of URF (subtype B and C). A total of 18 molecular clusters included 56 individuals (40.6%) were found in the transmission network under the optimal genetic distance threshold (1.0%). The clustering proportion of CRF07&#95;BC (66.1%, 37 cases) was higher than that of CRF01&#95;AE. There were 9 clusters formed among CRF07&#95;BC, including 37 cases (accounting for 61.7%, 37/60). The primary transmission cluster contained 11 cases, among which 9 cases were transmitted by homosexual sex. The first time of the cases to have homosexual behavior is range from 2010 to 2016, whose media number ( P (25), P (75)) of partners was 6 (3.5, 8.5). Most of the cases come from Anhui Province and engaged in garment industry (5 cases), between which there were 8 cases used Blued software to seek for casual partners, 1 case seeking for casual partners in garden. Conclusion: With CRF07&#95;BC and CRF01&#95;AE predominantly circulating, HIV genetic diversity had been noticed in this area. The primary cluster was consisted of high proportion of locally new infections, and a specific population aggregation in limited place existed.

Perilla frutescens,an annual plant in Labiatae family,is grown throughout China and can be used for medicine purposes and as food additives. The present field experiment was carried out to study the effects of different fertilizer treatments on the concentrations and accumulations of antioxidant components,including flavonoids and polyphenols,growth,seed yields and qualities of this plant.The main aim of this study is to provide farmers some advice for improving the yields and qualities of P. frutescens in theory and practice.Five treatments were set up,including a no fertilizer control(CK),chemical fertilizers(CF),organic fertilizers(M),organic fertilizers plus chemical fertilizers at the rates of 1 ∶1 and 1 ∶3 in terms of nitrogen(50 M,25 M). Plant growth parameters were recorded and total flavonoids and polyphenols were determined in three key growth stages of P. frutescens. At the fast growth period,samples of roots,leaves,and stems were collected for determining a total of flavonoids and polyphenols as well as DPPH removal rate of ethanol extracts. Seed yields and qualities were also recorded at harvest. The results showed fertilization enhanced growth and seed yields although no significant difference was observed in growth and seed yields in inorganic-organic fertilizer treatments. The total flavonoids,polyphenols,and DPPH removal rate of ethanol extracts followed the sequence leaves>stems>roots,indicating synthesis of these metabolites in the leaves. DPPH removal rate showed a positive linear correlation with total flavonoid and polyphenol concentrations. In addition,organic-inorganic fertilization significantly increased the numbers of both effective panicles and paniclegrains. Fertilizer treatments had no effect on seed qualities of P. frutescens,while 50 M achieved the highest yield,which increased by 14. 73% compared to CF alone. In general,50 M increased antioxidant components,biomass,and seed yield of P. frutescens,meriting advocate in cultivation.

Objective: To explore the psychological factors and blood supply to the penis in ED patients., Methods: From August to November 2018, we obtained Symptom Checklist 90 (SCL-90) scores and conducted an Eysenck Personality Questionnaire (EPQ) investigation among 106 ED patients in our Department of Andrology, followed by the test of intracavernosal injection of the vasoactive agent (ICI-VAA) and color duplex Doppler ultrasonography (CDDU). According to the results of ICI-VAA and CDDU, we divided the patients into a normal and an abnormal group, compared their item and total scores on SCL-90 with the national norm by t-test and analyzed the distribution of personality types in different groups by Chi-square test., Results: The results of ICI-VAA and CDDU showed 42 cases to be normal and the other 64 abnormal. Compared with the normal group, the abnormal cases exhibited significantly decreased peak systolic velocity (PSV) in the left and right corpus cavernosum (［30.90 ± 6.83］ vs ［19.39 ± 5.07］ cm/s, P < 0.01; ［30.74 ± 7.00］ vs ［18.98 ± 4.57］ cm/s, P < 0.01), but increased end-diastolic velocity (EDV) in the left corpus cavernosum (［1.77 ± 2.38］ vs ［3.21 ± 2.78］ cm/s, P < 0.01). The ED patients undergoing ICI-VAA and CDDU obtained remarkably higher SCL-90 scores than the national norm on depression (1.70 ± 0.64 vs 1.50 ± 0.59, P < 0.01), anxiety (1.62 ± 0.56 vs 1.39 ± 0.43, P < 0.01), compulsion (1.86 ± 0.61 vs 1.62 ± 0.58, P < 0.01), hostility (1.65 ± 0.66 vs 1.48 ± 0.56, P < 0.01), phobia (1.33 ± 0.45 vs 1.23 ± 0.41, P = 0.02), and psychoticism (1.61 ± 0.55 vs 1.29 ± 0.41, P < 0.01) as well as a higher total score (1.61 ± 0.48 vs 1.44 ± 0.48, P < 0.01). The patients with normal results of CDDU got markedly increased scores on anxiety (1.60 ± 0.53, P < 0.01), compulsion (1.83 ± 0.63, P = 0.02) and psychoticism (1.61 ± 0.48, P < 0.01), and so did those with abnormal results of CDDU on depression (1.73 ± 0.65, P < 0.01), anxiety (1.64 ± 0.59, P < 0.01), compulsion (1.88 ± 0.60, P < 0.01), hostility (1.68 ± 0.75, P < 0.01), phobia (1.35 ± 0.44, P = 0.02), psychoticism (1.61 ± 0.59, P < 0.01) and an increased total score as well (1.63 ± 0.51, P < 0.01). Statistically significant differences were not found in either the item scores or the total score on SCL-90 (P > 0.05), nor in the distribution of the EPQ personality types between the normal and abnormal cases (χ2 = 1.12, P = 0.77)., Conclusions: The personality types of ED patients are mainly phlegmatic and melancholic, and their psychological problems chiefly include depression and anxiety, which are more serious in those with insufficient penile blood supply.

Objective: Screen the pathogenic genes of a pedigree with clinical manifestation of familial dilated cardiomyopathy in Inner Mongolia. Methods: A total of 3 patients with dilated cardiomyopathy and 20 family members from the same family were examined in Ordos Central Hospital in Inner Mongolia from October, 2003 to August, 2017. Data on medical history, physical examinations, electrocardiograms, and echocardiography were obtained. 5 ml peripheral blood was sampled for per person. Chip Capture Sequencing technology was used to capture all the exons and splice sites of the genes that associated with hereditary cardiomyopathy and hereditary arrhythmia. The mutations in these genes were detected by high-throughput sequencing. All suspected pathogenic loci identified by high-throughput sequencing were verified by Sanger sequencing used for mutation detection. One hundred and fifty gender, age and race matched healthy people were included as the control group. Results: Pathogenic gene variations were detected in 3 symptomatic family members and 1 carrier from the pedigree. Five pathogenic gene variations were identified in the proband (Ⅱ1), a pSer236Gly and a pArg215Cys variation in the MYBPC3 gene, a pGln90Arg variation in the DSP gene, and pAsn2912Asp and pGlu2910Val variation in the DMD gene. One pathogenic variation was detected in Ⅲ3, which was a pArg215Cys variation in the MYBPC3 gene. Two pathogenic variations were detected in Ⅲ7, a pSer236Gly variation in the MYBPC3 gene and a pGln90Arg variation in the DSP gene. Two pathogenic variations were detected in the Ⅳ7, a pSer236Gly variation in the MYBPC3 gene and a pGln90Arg variation in the DSP gene. No gene variation loci were detected in the other family members and the control group. Conclusion: MYBPC3 gene, DSP gene and DMD gene variations are present in the familial dilated cardiomyopathy pedigree from Inner Mongolia, and these variations may be related with familial dilated cardiomyopathy.

Objective: To explore the role of miR-593 in regulating the proliferation of colon cancer cells and the molecular mechanism., Methods: Bioinformatics analysis identified PLK1 as the possible target gene of miR-593. Luciferase assay was employed to verify the binding between miR-593 and PLK1, and qRT-PCR and Western blotting were used to verify that PLK1 was the direct target gene of miR-593. CCK-8 assay was performed to test the hypothesis that miR-593 inhibited the proliferation of colon cancer cells by targeting PLK1., Results: Luciferase assay identified the specific site of miR-593 binding with PLK1. Western blotting showed a significantly decreased expression of PLK1 in the colon cancer cells transfected with miR-593 mimics and an increased PLK1 expression in the cells transfected with the miR-593 inhibitor as compared with the control cells ( P &lt; 0.05). The results of qRT-PCR showed no significant differences in the expression levels of PLK1 among the cells with different treatments ( P &gt; 0.05). The cell proliferation assay showed opposite effects of miR-593 and PLK1 on the proliferation of colon cancer cells, and the effect of co-transfection with miR-593 mimic and a PLK1-overexpressing plasmid on the cell proliferation was between those in PLK1 over-expressing group and miR-593 mimic group., Conclusions: miR-593 inhibits the proliferation of colon cancer cells by down-regulating PLK1 and plays the role as a tumor suppressor in colon cancer.

Objective: To understand the epidemiological characteristics of one large HIV molecular transmission cluster in Jiaxing city, Zhejiang province, 2017 in order to select those people under high-risk and providing basis for programs on prevention. Methods: During 2017, newly diagnosed HIV/AIDS cases in this city were recruited. Plasma samples were collected from subjects, followed by RNA extraction, RT-PCR and nest-PCR for pol gene amplification, before being sequenced and aligned. Mega 6.0 software was used to construct phylogenetic tree, and Cytoscape 3.6.0 software was used to identify HIV molecular transmission clusters. Cases within the large transmission clusters were investigated, using a field-epidemiology-questionnaire. Data related to socio-demographics and previous sexual behaviors were collected and EpiData 3.0 and SPSS 20.0 software were used. Results: In the large transmission cluster with subtype identified as CRF07&#95;BC, in Jiaxing, 2017, 26 cases of the total 30 cases were investigated. A total of 80.8% (21/26) could be identified as newly infected within the last two years and 30.8%(8/26) could be identified as newly infected within the last one year, including 22 cases infected locally. Among several infected cases who were at age 45 years or older, they admitted that they had experienced unprotected sexual contacts in local city for long time and having had more than 10 disclosed sexual contacts within the last two years at the local venues. Conclusions: This molecular cluster had been formed and scaled up quickly in recent two years, it has played an important role in promoting and scaling up the HIV transmission. Three cases identificed as high risk played an importantrde role in scaling up this cluster.

Modern pharmacological studies have shown that Shengmai San has the effects of enhancing immunity and improving blood circulation, and Curcumae Longae Rhizoma(Jianghuang) has anti-inflammatory, anti-cancer, anti-oxidation and other functions. Shengmai San combined with Jianghuang is a new research direction in the study of anti-tumor of traditional Chinese medicines. The main treatment for nasopharyngeal carcinoma is radiation therapy, but radiation therapy can cause a variety of side effects, and it also changes the composition of the intestinal flora. In this study, the 16 s rDNA sequencing platform was used to perform macro-sequence sequencing of the intestinal flora samples of nude mice bearing the veins of Shengmai Jianghuang San, and then the results of intestinal flora data were analyzed to investigate the effect of Shengmai Jianghuang San on tumors. The results showed that Shengmai Jianghuang San combined with irradiation could enhance the therapeutic effect of tumor treatment. Radiation therapy would reduce the total number and diversity of intestinal flora in nude mice, and also change the structure of the flora. Shengmai Jianghuang San could protect the diversity of colonies, and also partially restore the colony imbalance caused by irradiation. This study provides a research idea for Shengmai Jianghuang San as a sensitizing adjuvant for radiotherapy of nasopharyngeal carcinoma.

Objective: To investigate the role of miR-144-3p in regulating osteogenic differentiation of bone marrow mesenchymal stem cells and predict its target genes., Methods: Rat bone marrow mesenchymal stem cells (BMSCs) with induced osteogenic differentiation were examined for the expressions of Runx2, OCN and miR-144-3p. The effects of transfection with a miR-144-3p mimic or a miR-144-3p inhibitor were tested on the osteogenic differentiation of the BMSCs. The changes in the expressions of the predicted target of miR-144-3p in the BMSCs during induced osteogenic differentiation were examined using Western blotting and qRT-PCR., Results: Rat BMSCs with induced differentiation into osteoblasts exhibited a progressive increase in the expressions of Runx2 and OCN (two markers of osteogenic differentiation), while the expression of miR-144-3p gradually decreased during the differentiation till reaching the lowest level at 21 days of induction. In rat BMSCs, transfection with the miR-144-3p mimic significantly decreased ALP activity ( P &lt; 0.05) wile transfection with the miR-144-3p inhibitor significantly increased ALP activity ( P &lt; 0.05) in rat BMSCs. Analysis based on miRanda, microRNA.org database and TargetScan suggested that Smad4 was the most likely target gene of miR-144-3p. The results of qRT-PCR showed no significant differences in expression levels of Smad4 among the cells with different treatments ( P &gt; 0.05), while Western blotting revealed a significantly decreased expression of Smad4 in the cells transfected with miR-144-3p mimics and an increased Smad4 expression in the cells transfected with the miR-144-3p inhibitor as compared with the control cells ( P &lt; 0.05)., Conclusions: miR-144-3p participates in the regulation of osteogenic differentiation of rat BMSCs, and its inhibitory effect on osteogenic differentiation is achieved probably by decreasing the expression level of Smad4.

Objective: To understand the molecular subtypes and recombinant structure of HIV-1 infection in Zhejiang Province. Methods: According to the method of stratified random sampling method, a total of 302 newly HIV infected people (excluding AIDS patients) were selected as the subjects of the investigation from April to June, 2015, who were all newly HIV positive patients reported in Zhejiang Province and were diagnosed as HIV positive and had not received antiretroviral treatment. Questionnaires were used to collect information such as gender, residency, marital status, route of transmission and case reporting area. At least 8 ml of venous blood was collected from HIV infected people, reverse transcription-polymerase chain reaction (RT-PCR) and nested PCR were used to amplify the pol gene sequence of HIV-1 strains and circulating recombination forms (CRF) and unique recombinant form (URF) strains were screened using phylogenetic tree, and molecular subtype analysis and recombinant analysis were performed. Results: Of the 302 HIV infected people, 18 cases were infected with new CRFs or URFs, among which 15 were male (83.3%), 3 were heterosexual transmission (17.7%); 8 cases (44.4%) were infected with new CRFs, which included CRF55&#95;01B 5(27.8%), CRF67&#95;01B 2 (11.1%), CRF69&#95;01B 1 (5.6%); 10 cases (55.6%) were infected with URFs, in which 6 cases were infected with two groups of recombinant strains composed of CRF01&#95;AE subtype and CRF07&#95;BC subtypes in pol region, 3 cases were infected with three groups of recombinant strains of the two subtypes in pol region, and 1 case could not be accurately determined of the recombinant structure. The results of recombinant source analysis showed that the newly recombinant strains found from Zhejiang province were highly homologous to the strains from Guangdong, Anhui, Hebei, Yunnan, Liaoning province, and the sequence similarity was 94.4%-98.0%. Conclusion: The new recombinant strains of Zhejiang province included CRFs strains and URFs strains with CRF01&#95;AE and CRF07&#95;BC as the main recombinant structure mode, and all of them were introduced from other places and formed a trans-regional chain of HIV transmission.

Objective: To understand the characteristics of distribution on HIV-1 subtypes and the transmission clusters in Yiwu in Zhejiang province. Methods: A cross-sectional study of molecular epidemiology was carried out on newly reported HIV/AIDS cases in Yiwu. RNA was extracted from 168 plasma samples, followed by RT-PCR and nest-PCR for pol gene amplification, sequencing, phylogenetic tree construction used for analyzing the subtypes and transmission clusters. Mutations on drug resistance was analyzed by CPR 6.0 online tool. Results: Subjects were mainly males (86.3%, 145/168), with average age as (39.1±13.4) years old and most of them were migrants (66.7%, 112/168). The major routes of transmission included homosexual (51.2%, 86/168) and heterosexual (48.8%, 82/168) contacts. The rate of success for sequence acquisition was 89.9% (151/168). The dominant subtypes showed as CRF01&#95;AE (74, 49.0%) and CRF07&#95;BC (64, 42.4%), followed by CRF08&#95;BC (5, 3.3%), CRF55&#95;01B (3, 2.0%), each case of subtype B, CRF45&#95;cpx, CRF59&#95;01B, CRF85&#95;BC and URF (B/C). CRF45&#95;cpx and CRF85&#95;BC were discovered the first time in Zhejiang province. Twenty-six transmission clusters involving 65 cases were found, with the total clustered rate as 43.0% (65/151), in which the CRF01&#95;AE clustered rate appeared as 54.1% (40/74), higher than that of CRF07&#95;BC (21/64, 32.8%). The average size of cluster was 2.5 cases/cluster, with average size of cluster in CRF01&#95;AE patients infected through heterosexual transmission as the largest (3.5 cases/cluster). The prevalence of transmitted drug resistance was 4.6% (7/151). Seven cases with surveillance drug resistant mutations (SDRM) were found, including 5 cases of M46L (3.3%), and one case of F77L or Y181C. Conclusion: HIV genetic diversity and a variety of transmission clusters had been noticed in this study area (Yiwu). Programs on monitoring the subtypes and transmission clusters should be continued and strengthened.

Objective: To compare the efficacy differences between ginger-separated moxibustion at Baliao points combined with Bushen Huoxue formula and Bushen Huoxue formula alone on patients with decreased ovarian reserve function., Methods: Fifty patients of decreased ovarian reserve function were randomly divided into an observation group and a control group, 25 cases in each one. The patients in the observation group were treated with ginger-separated moxibustion at Baliao points combined with Bushen Huoxue formula; the moxibustion was given for 1.5 h, once every seven days, and no treatment was given during menstrual period. The patients in the control group were treated with Bushen Huoxue formula. One-month treatment was taken as one treatment course, and totally three courses were given. The change of follicle-stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E 2 ), anti-mullerian hormone (AMH), antral follicle count (AFC), peak systolic velocity (PSV), resistance index (RI) were observed before and after treatment in the two groups., Results: After treatment, the FSH, FSH/LH and RI were significantly lowered, but the E 2 , AFC, PSV were significantly increased in the two groups (all P <0.05); the FSH, FSH/LH and E 2 in the observation group were lower and AFC was higher than those in the control group (all P <0.05)., Conclusion: The ginger-separated moxibustion at Baliao points combined with Bushen Huoxue formula are superior to Bushen Huoxue formula alone in improving ovarian reserve function.

Objective: To study the regulation of sporulation controlled by two-component system (TCS) YvcPQ., Methods: β-galactosidase experiment was used to verify the regulation of YvcP on kapD expression; bacterial one-hybrid assay, EMSA and RT-qPCR were applied to study the regulation of AbrB on yvcPQ expression; markerless gene deletion coupled with spore count was used to reveal the influence of yvcPQ and kapD expressions on sporulation., Results: transcriptional regulator AbrB up-regulated the expression of yvcPQ; YvcP promoted the expression of kapD to inhibit sporulation., Conclusion: AbrB up-regulated the transcription of yvcPQ operon, then the increased YvcP strengthened the transcriptional acitivation of sporulation inhibitor gene kapD, and subsequently inhibited sporulation.

Objective: To investigate the clinical effect of Jujing No.2 Decoction on male infertility induced by varicocele., Methods: We equally randomized 70 male patients with varicocele-induced infertility into a treatment group and a control group, the former treated with modified Jujing No.2 Decoction (1 dose a day, bid) and the latter given Danshen Tablets (4 tablets once, tid) plus Liu Wei Di Huang Wan (8 pills once, tid), both for 3 months. We examined the spermatic veins of the patients by ultrasonography and performed seminal analysis before and after medication., Results: Compared with the controls, the patients in the treatment group showed significant improvement after medication in sperm concentration (［12.35±2.12］ vs ［18.56±4.16］ ×10⁶/ml, P<0.05), progressively motile sperm (［16.18±6.34］ vs ［24.63±8.51］ %, P<0.05), and morphologically normal sperm (［11.59±5.31］ vs ［14.17±6.02］ %, P<0.05). In the control group, sperm concentration was increased from (12.29±2.07) ×10⁶/ml at the baseline to (13.13±3.13) ×10⁶/ml after medication, progressively motile sperm from (16.13±6.45) % to (20.55±7.30) % (P<0.05), and morphologically normal sperm from (11.62±5.35) % to (12.69±5.77) %. The total effectiveness rate was significantly higher in the treatment than in the control group (71.88% vs 46.67%, P<0.05). The inner diameter of the left spermatic vein was decreased from (0.32±0.05) mm at the baseline to ［0.26±0.05］ mm after medication in the treatment group (P<0.05) and from (0.32±0.03) mm to (0.29±0.04) mm in the control (P<0.05), with statistically significant difference between the two groups after medication (P<0.05)., Conclusions: Jujing No.2 Decoction is effective in the treatment of varicocele-induced male infertility.

4-coumarate coenzyme A ligase is a key enzyme of phenylpropanoid metabolic pathway in higher plant and may regulate the biosynthesis of ferulic acid in Angelica sinensis. In this study, the homology-based cloning and rapid amplification of cDNA ends (RACE) technique were used to clone a full length cDNA encoding 4-coumarate coenzyme A ligase gene (4CL), and then qRT-PCR was taken for analyzing 4CL gene expression levels in the root, stem and root tissue at different growth stages of seedlings of A. sinensis. The results showed that a full-length 4CL cDNA (1,815 bp) was obtained (GenBank accession number: KT880508) which shares an open reading frame (ORF) of 1 632 bp, encodes 544 amino acid polypeptides. We found 4CL gene was expressed in all tissues including leaf, stem and root of seedlings of A. sinensis. The expressions in the leave and stem were increased significantly with the growth of seedlings of A. sinensis (P < 0.05), while it in the root showed little change. It indicates a time-space pattern of 4CL gene expression in seedlings of A. sinensis. These findings will be useful for establishing an experiment basis for studying the structure and function of 4CL gene and elucidating mechanism of ferulic acid biosynthesis and space-time regulation in A. sinensis.

Objective: To evaluate the success rate of dental implants and the changes of sinus membrane thickness after sinus lift in cases of thickened sinus membrane., Methods: Sixteen patients without maxillary sinusitis and with 5-8 mm residual alveolar bone heights were included in this study. The sinus membrane thickeness of these patients were more than 2 mm. All patients received sinus lift surgery and dental implants insertion. The changes of sinus membrane was evaluated by cone-beam CT (CBCT) pre-surgery and 6 months after sinus lift surgery, and the short term success rate of dental implants was also evaluated., Results: A total of 18 implants from 16 subjects were inserted. The thickness of membrane was decreased in 14 cases after sinus lift and increased in 2 cases. All the orifice of maxillary sinus was unobstructed before surgery, one case was obstructed after surgery without inflammation. All the dental implants succeeded., Conclusions: Sinus augmentation with thickened sinus membrane is not contraindication of sinus lift.

Carbon and nitrogen removal performance and microbial community structure under different dissolved oxygen (DO) conditions (3, 2, 1 and 0. 5 mg . L -1) in an anoxic/oxic (A/O) system were investigated. The results showed that the A/O activated sludge system still had an excellent performance in removing carbon and nutrient under low DO condition (0. 5 mg . L-1). The removal rates of chemical oxygen demand (COD), ammonia (NH4+ -N) and total nitrogen (TN) were 89. 7%, 98. 3% and 88. 0% respectively. The PCR-DGGE analysis showed that the bacterial community structure changed greatly under different DO conditions. However, there was still a high bacterial diversity even at low DO level, which ensured the functional stability of the A/O system. On the basis of the results of the phylogenetic tree, bacterial communities were observed to be very abundant, and Proteobacteria was identified as the dominant bacteria.

Objective: To investigate the clinicopathologic characteristics of epithelioid hemangioendothelioma (EHE)., Methods: Thirteen cases of EHE were analyzed by gross examination, light microscopy and immunohistochemical staining., Results: The patients included 6 males and 7 females. The age of patients ranged from 11 to 74 years (mean = 43 years). The tumors were located in soft tissue, liver, lung, mediastinum, sacrum, and pleura. The tumor cells were epithelioid, spindled or polygonal-shaped and exhibited cord-like or small nest growth pattern. Some tumor cells formed cytoplasmic vacuoles, which contained erythrocytes. The tumor cells showed abundant eosinophilic cytoplasm. The stroma varied from highly myxoid to hyaline. In 3 cases, the tumors contained areas with significant atypia, brisk mitotic activity and necrosis. Immunohistochemical study showed that the tumor cells were positive for CD31, ERG, FLI-1, CD34, factor VIII and CKpan to different extents. Follow-up information was available in 10 patients with duration ranging from 8 to 65 months. Seven patients were alive and three died., Conclusions: EHE is easily a rare maligant vascular tumor,occurs in many organs other than soft tissue.EHE is misdiagnosed as carcinoma for its significant epithelioid morphology. Recognizing its clinicopathologic characteristics and combined application of specific vascular endothelial immunohistochemical markers are important to avoid confusion with other lesions.

Objective: To investigate the impacts of specific serines phosphorylation of αA Crystallin on anti-oxidation and anti-apoptotic ability of human lens epithelial cells (HLEC)., Methods: Experimental Study. Using site-directed mutagenesis, serines on specific sites (Ser45, Ser66, Ser122) were replaced respectively by aspartic acid (D) to mimic phosphorylation. Target genes were inserted into pEGFP-C1 plasmid and transfected into HLEC by Lipofectamine™ 2000. HLEC transfected with blank plasmid and plasmid over-expressing wide-type (WT) αA Crystallin were defined as control group and WT group. After H2O2 induction, Dihydroethidium (DHE) and Annexin V-PE/7AAD staining were used to detect the reactive oxygen species level and apoptotic rate. T-test, Wilcoxon-two sample test, analysis of variance and Kruskal-Wallis test were used for statistical analysis., Results: The DHE intensity of three phosphorylation groups were 167.53±18.21, 143.54±4.78 and 143.40±30.19, which reduced significantly comparing to control group (274.42±3.71, t=9.963, P=0.001; t=34.961, P<0.01; Z=1.993, P=0.046); The same tendency were found when compared with cell group (WT) over-expressing wide-type αA Crystallin (192.67±1.47, Z=1.964, P=0.049; t=17.887, P<0.01; Z=1.964, P=0.049). After H2O2 induction for apoptosis, early apoptotic rates for three phosphorylation groups were 15.08%±0.90%, 16.59%±5.37% and 15.36%±2.62% respectively, which significantly decreased comparing with control group (41.28%±2.70%; t=21.632, P<0.01; t=8.855, t=15.450, P<0.01) and WT group (31.40%±1.64%, t=15.117, P<0.01; t=4.524, P=0.006; t=8.988, P=0.001). No significant differences were found in both measurements between the three phosphorylation groups (F=1.08, P=0.407; x2=4.345, P=0.114)., Conclusion: The Ser45, Ser66 and Ser122 phosphorylation enhanced the anti-oxidative stress and anti-apoptotic capability of HLEC.

Objective: To evaluate the effect of "one village, one control strategy" on schistosomiasis control in Qianjiang City., Methods: The villages with human and/or bovine schistosome infection rate being more than 1.5% in 2011 were chosen as the trial villages. According to the epidemic characteristics of schistosomiasis and control difficulties of the villages, the "one village, one control strategy" was performed and the control effects were evaluated., Results: By the end of 2013, the infection rates of human and bovine in the trial villages decreased by 45.91% (χ2 = 469.19, P < 0.01) and 49.38% (χ2 = 11.46, P < 0.01), respectively, and all the infection rates of human and bovine in those villages were less than 1%., Conclusion: The "one village, one control strategy" is effective for schistosomiasis control, which can rapidly reduce the prevalence of schistosomiasis.