DNA barcoding has been widely used for herb identification in recent decades,enabling safety and innovation in the field of herbal medicine.In this article,we summarize recent progress in DNA bar-coding for herbal medicine to provide ideas for the further development and application of this tech-nology.Most importantly,the standard DNA barcode has been extended in two ways.First,while conventional DNA barcodes have been widely promoted for their versatility in the identification of fresh or well-preserved samples,super-barcodes based on plastid genomes have rapidly developed and have shown advantages in species identification at low taxonomic levels.Second,mini-barcodes are attractive because they perform better in cases of degraded DNA from herbal materials.In addition,some mo-lecular techniques,such as high-throughput sequencing and isothermal amplification,are combined with DNA barcodes for species identification,which has expanded the applications of herb identification based on DNA barcoding and brought about the post-DNA-barcoding era.Furthermore,standard and high-species coverage DNA barcode reference libraries have been constructed to provide reference se-quences for species identification,which increases the accuracy and credibility of species discrimination based on DNA barcodes.In summary,DNA barcoding should play a key role in the quality control of traditional herbal medicine and in the international herb trade.

Objective: To analyze the genetic characteristics of the hemagglutinin(HA) and neuraminidase(NA) genes of the influenza A/H1N1(09pdm) viruses isolated in the city of Yancheng in 2014-2017. Methods: The throat swab specimens of patients with influenza-like illness (ILI) from sentinel surveillance hospitals and outbreak sites were detected using the method of real time RT-PCR. The influenza A/H1N1(09pdm) viruses were isolated using MDCK cell culture method in 2014-2017. The strains in 2014-2017 were selected randomly and their sequences of the HA1 and NA genes were amplified through one step RT -PCR method and the PCR products were sequenced. The mutations of genes and acid locus were analyzed and the evolutional trees were generated using bioinformatics software. Results: The clustering relationships of the respective branches of HA1 and NA genes of seventeen A/H1N1(09pdm) strains isolated in Yancheng area were basically the same and the phylogenetic trees of HA1 and NA genes were respectively clustered into four evolutionary branches. Compared with the vaccine strain A/California/07/2009(H1N1pdm)in the Northern Hemisphere, a total of three antigen epitopes (Ca, Sa, Sb) in HA1 genes of strains in Yancheng area were involved in six antigenic sites (K154R, S162N, K163Q, S185T, L191I, S203T); there were three mutations (D222G/N, G223R, E224K) in the 220 ring and one locus (L191I) in the 190 helix of the receptor binding sites; the two strains (A/Jiangsu-YC/SWL1540/2017, A/Jiangsu-YC/SWL1545/2017) isolated in 2017 increased the 162NQS glycosylation site. Because the strains of the antigen epitopes, receptor binding sites and glycosylation sites in the HA1 genes had a certain degree of variations in Yancheng area in 2014-2017, the protective effects of vaccine strain A/California/07/2009 (H1N1pdm) was limited at the gene level. The two strains (A/Jiangsu-YC/SWL1540/2017 and A/Jiangsu-YC/SWL1545/2017) isolated in 2017 were clustered with vaccine strain A/Michigan/45/2015(H1N1pdm) and had better protective effects. Seventeen A/H1N1(09pdm) strains had no mutations in catalytic residues and drug resistant sites of NA genes, but a part of strains had a certain degree of variations in glycosylation sites of NA genes. Conclusions These results indicated the HA1 and NA genes of influenza A/H1N1(09pdm) viruses circulated in Yancheng area in 2014-2017 changed gradually. The accumulation of these mutations would result in antigenic drift of influenza A/H1N1(09pdm) viruses.

Objective: To analyze the genotypes and the genetic evolution of the hemagglutinin genes of measles viruses in the city of Yancheng in 2016. Methods: Using a set of primers and probes for screening positive for measles viruses, specimens of throat swab were detected using the method of real time RT-PCR. The sequences of the nucleoprotein and hemagglutinin genes of measles viruses were amplified through one step RT-PCR method and the PCR products were sequenced. The sequences of nucleotide and amino acid of the nucleoprotein and hemagglutinin genes of measles viruses were analyzed and the evolutional trees were generated using bioinformatics software. Results: The genotypes of measles viruses in the Yancheng area in 2016 included subgenotype H1a and genotype D8. Phylogenetic trees analysis showed that the five representative strains of subgenotype H1a in Yancheng area and Jiangxi representative strain (KJ136545) clustered into independent evolutionary branches, belonged to the clade of H1a -1 evolutionary genes. The seven representative strains of genotype D8 in Yancheng area were clustered with the American representative strain in 2009 (JN635404), belonged to the D8-3-2 small clade genes. Compared with vaccine strain of Shanghai S191, the amino acid site in 240^thof the five representative strains of subgenotype H1a in Yancheng area mutated from serine to asparagine, leading to a loss of the N-glycosylation site NLS_238-240. The seven representative strains of genotype D8 in Yancheng area had no change in N-glycosylation. Conclusions In 2016, the prevalent strains of measles viruses in Yancheng area were mainly Chinese H1a dominant subgenotype and D8 imported genotype. In addition to a loss of the N-glycosylation site NLS_238-240in 240^thof the five representative strains of subgenotype H1a, most of the major neutralizing antigen sites of hemagglutinin gene of measles viruses in Yancheng area did not mutate. The Chinese vaccine of Shanghai S191 can effectively prevent infection caused by subgenotype H1a and subgenotype D8 strains.

Background and purpose:Laparoscopic retroperitoneal adrenalectomy makes access to the adrenal glands easier and less invasive than open surgery. The aim of this study was to evaluate the clinical efifcacy of laparoscopic retroperitoneal adrenalectomy.Methods:A total of 130 patients who underwent retroperitoneal adrenalectomy for adrenal mass from Jan. 2007 to Dec. 2012 in Fudan University Shanghai Cancer Center were retrospectively assessed. Their clinicopathological factors, perioperative complications and short-term prognostic data were retrieved from the medical records.Results:One hundred and twenty-seven of 130 patients underwent retroperitoneal adrenalectomy successfully, and 3 patients were converted to open surgery due to severe bleeding. Among 130 patients, 63 were male and 67 were female, with the mean age 50.0 years. The pathological results of the 130 patients indicated adrenocortical adenoma in 68, pheochromocytoma in 15, medullary lipoma in 13, adrenal cysts in 10 , ganglioneuroma in 7, metastatic cancer in 5, adrenal hyperplasia in 4, schwannoma in 3, lymphangioma in 2, adrenal hematoma in 1, adrenal cortical carcinoma in 1, adrenal angiosarcoma in 1 and the deputy spleen in 1 (one patient suffering from both pheochromocytoma and ganglioneuroma). The maximum diameters were ranging from 0.5 to 9.0 cm, and mean diameter was 3.48 cm. The average blood loss in surgery was 62.73 mL. Mean length of stay in hospital was 7 d. GradeⅠ complications occurred in 5 patients, including 2 of fever, 1 of food allergy, 1 of drug allergy and 1 of hypokalemia.Conclusion: Retroperitoneal adrenalectomy should be considered as the procedure of choice for the resection of most adrenal tumors in skilled centers with the advantages of minimal invasion, increased safety and faster recovery.

In this paper, the chloroplast psbK-psbI intergenic spacers of 18 species of Dendrobium and their adulterants were amplified and sequenced, and then the sequence characteristics were analyzed. The sequence lengths of chloroplast psbK-psbI regions of Dendrobium ranged from 474 to 513 bp and the GC contents were 25.4%-27.6%. The variable sites were 71 while the informative sites were 46. The inter-specific genetic distances calculated by Kimura 2-parameter (K2P) of Dendrobium were 0.006 1-0.058 1, with an average of 0.028 4. The K2P genetic distances between Dendrobium species and Bulbophyllum odoratissimum were 0.093 2-0.120 4. The NJ tree showed that the Dendrobium species can be easily differentiated from each other and 6 samples of the inspected Dendrobium species were identified successfully through sequencing the psbK-psbI intergenic spacer. Therefore, the chloroplast psbK-psbI intergenic spacer can be used as a candidate marker to identify Dendrobium species and its adulterants.

There is a high degree of information asymmetry in medical services.Public hospitals are characteristic of the nature of multi-layer principal-agent relationships.Asymmetric information can lead to inconsistent value goals between the principal and the agent,giving birth to the government regulation capture as a result.Such capture ranks a fundamental cause for diluted public welfare nature of public hospitals.To offset such capture at public hospitals calls for reconstruction of government regulation system within the framework of public hospital corporate governance at an institution level.It is also necessary to eliminate the information asymmetry and its negative results,bringing public hospitals back on track to public welfare.

Objective To investigate the effect of Terpinen-4-ol on the proliferation of lung adenocarcinoma A-549 cells and its related mechanism. Methods A-549 cells were treated with different concentrations of Terpinen-4-ol. The inhibitory effect of Terpinen-4-ol on A-549 cells was tested by MTT method. Cell grow ability was determined by CCK-8 colorimetry. The ultrastructure of A549 cells were observed by transmission electron microscopy before and after Terpinen-4-ol treatment. The changes of cell cycle, apoptosis, and the level of intracel-lular calcium were inspected by flow cytometry. Inoculated the lung adenocarcinoma A-549 cells on the nude mice to form transplantation tumor. The experimental nude mice with transplantation tumors were divided into three groups:negative control group,high dose positive con-trol group and low dose positive control group. The mice were given continuously intraperitoneal injection for 10 days, and then the transplan-tation tumors were taken and the size and weight of them were detected. Results After Terpinen-4-ol treatment for 24 h,MTT assay showed that the IC50 value of A549 cells was 0. 067% v/v. The growth curves of positive control groups were significantly smooth than the negative control group. The formation of autophagosome increased after treatment with Terpinen-4-ol. The results of flow cytometry showed that the cell cycle was arrested in S phase,Terpinen-4-ol could induce apoptosis of A549 cell, The intracellular calcium concentrations in positive control groups were significantly higher than the negative control group(P<0. 05). Low dose group and high dose group restrained the growth of the transplantation tumor obviously, and the tumor inhibitory rate were 53. 33% and 77. 76% respectively. Conclusion Terpinen-4-ol has inhibitory effect on the proliferation of A-549 cells in vitro and in vivo.

Objective To validate the Memorial Sloan-Kettering Cancer Center(MSKCC)score model and evaluate the clinical efficacy of vascular endothelial growth factor(VEGF)-targeted agents in the treatment of advanced renal cell carcinoma(RCC)in China.Methods Three hundred and forty-five patients with advanced RCC and average age of 57(17-90)years were treated with VEGF-targeted agents.There were 306 cases of clear cell RCC,20 cases of papillary RCC,4 cases of chromophobe RCC,5 cases of renal collecting duct carcinoma,3 cases of medullary carcinoma and 7 cases of unclassified RCC.The main metastatic lesions were located at lung,bone and lymph nodes.Of them,205 cases were given the treatment of sorafenib 400 mg bid without off treatment,while 140 cases received sunitinib treatment in repeated six week cycles consisting of four weeks of sunitinib 50 mg daily followed by two weeks off treatment.Overall survival(OS)was estimated by the Kaplan-Meier method.Log-rank test and Harrell concordance index analysis were used to validate the MSKCC score model.Results The median follow-up period were 23(1-68)months in the whole group.The OS was 33 months,and survival rates at 1,2,3 year were 77.6％,59.3％,46.6％,respectively.According to the MSKCC score model,the patients were segregated into three risk categories: the favorable-risk group(no prognostic factors;n =169;49.0％),in which median OS(mOS)was 46 months and 2 year OS was 75.8％;the imtermediate-risk group(one or two prognostic factors;n =150;43.5％),in which mOS was 24 months and 2 year OS was 47.7％;and the poorrisk group(three to five prognostic factors;n =26;7.5％),in which mOS was 8 months and 2 year OS was 10.1％(log-rank P ＜ 0.01).The concordance index was 0.687.Conclusions VEGF-targeted agents are effective in Chinese advanced RCC patients.The MSKCC score model can be incorporated into judging individualizing tumor prognosis and communicating about the treatment options with patients who are using VEGF-targeted agents.

Medicinal animals are important part of Traditional Chinese medicine resources in China. Cytochrome c oxidase subunit I (COI) was selected as the standard DNA barcoding sequence for animal medical materials. In this study, the 51 animal species from 45 animal medical materials in the Chinese Pharmacopoeia were selected and the intra-specific variation and the inter-specific divergence, the barcoding gap, the identification efficiency of their COI sequences were analyzed. The results showed that the inter-specific divergence is higher than intra-specific distance. The barcoding gap existed between inter-specific sequence divergence and intra-specific dis-tance. The identification efficiencies were 100% both at the genus and species level except the Arthropoda. The cluster dendrogram exhibited that different species distinguished from others. Therefore, COI sequence as a bar-code is suitable to identify the species of animal medical materials in Chinese Pharmacopoeia.

Background and purpose: Platelet to lymphocyte ratio (PLR) is an important factor reflected systematic inflammation. The clinical value of PLR has not been confirmed. The present study was to explore the value of preoperative PLR in predicting clinical stage and prognosis in upper tract urothelial carcinoma. Methods:Patients who underwent surgical therapy with postoperative pathology upper tract urothelial carcinoma without metastasis from Jan. 2007 to Mar. 2012, were collected. Following up was done by telephone and clinic work, 150 vs 1 was taken as the threshold value of PLR, and the association of PLR with tumor stage, whether suffered bladder cancer as comorbidity, recurrent or metastasis, overall survival, tumor lesion, preoperative hematuria, gender and age was analyzed. We further analyzed the association difference of disease free survival (DFS) time and overall survival (OS) time between different PLR groups. Results:Fifty-one cases of UTUC were collected, and the postoperative mean following up time is 21 (9–51) months. Twenty cases recurred or metastasis and 9 cases died. The mean DFS time was 15 (2–51) months,and the mean OS time was 21 (9–51) months. One-factor analysis of variance showed that preoperative PLR was associated with tumor stage, overall survival rate, hematuria and gender, and the P value were 0.028, 0.008, 0.045, 0.036 respectively. High PLR group was intended to be non-organ confined disease, the sensitivity was 57%and the specificity was 74%. Survival analysis by Kaplan-Meier method showed there is no statistical difference in DFS between high and low PLR groups (P=0.155). But OS time in high PLR group was significantly less than that in low PLR group (P=0.006). Cox regression confirmed that only tumor stage is an independent prognostic factor of OS (P=0.029). Conclusion:PLR has potential clinical value in predicting advanced stage disease and Cox regression confirmed that only tumor stage is an independent prognostic factor of OS.