Objective:To observe the therapeutic effect and mechanism of Huzhangyuzhuo decoction on treating chronic pelvic pain syndrome(CPPS).Methods:46 male Wistar rats were randomly divided into control group(n=10) and experiment group(n=36).The CPPS rat model was induced by intraperitoneal injection of diphtheria-pertussis-tetanus vaccine and subcutaneous injection of puri ed prostate protein with FCA suspension.Rats in experiment group were subdivided into model group,Chinese medicine group and western medicine group(n=12).After 30 days of treatment,pathology changes of prostate were observed,and levels of IL-1?,COX-2,PGE2,?-EP in prostate homogenate were measured by ELISA.Results:Huzhangyuzhuo decoction greatly reduced the in ammation of prostate and relieved the structure damage.At the same time,Huzhangyuzhuo decoction can decrease the level of IL-1?,COX-2,PGE2 and increase level of ?-EP.Conclusions:Huzhangyuzhuo decoction might treat CPPS by regulating of cytokines in the prostate,depressing in ammation and relieving structure damage.

Objective: In China, doctors at TCM hospitals and clinics often divide patients with a Western medicine (WM) disease into several syndrome classes from the TCM perspective and treat patients in different classes using different principles. A key problem is how to carry out the classification properly. We propose an evidence-based ap-proach for solving the problem where evidence is obtained by analyzing unlabeled symptom data using latent tree models.Method: In previous work, we have shown how latent tree analysis of symptom data can be used to identify TCM syndrome classes among patients with a WM disease. In the paper, we investigate how to establish classification rules for distinguishing between the classes.Results: We have applied the method to a data set about Vascular Mild Cognitive Impairment that involves 93 symptoms and 803 patients. Nine syndrome types are identified, along with the corresponding classification rules. Conclusions: An evidence-based approach to the TCM patient classification prob-lem has been developed. The approach can be used to answer the following questions about a WM disease: What TCM syndrome classes are there? What are the sizes of the classes? What are the statistical characteristics of each class? How can one differentiate between the different classes?

Objective To construct Cox7a2 fluorescent vector and study its effect on cytochrome C oxidase (COX) activity in mouse Sertoli cell line TM4. Methods The coding region of Cox7a2 was amplified from mouse Sertoli cell line TM4 by RT-PCR. The PCR product was inserted into pEYFP-C1 vector with BamH I and EcoR I restriction site, and confirmed by DNA sequencing. The recombinant fusion protein vector was amplified by transforming into DH5a and transfected into TM4 cells. The protein expression was identified by Western blot. COX activity was measured by spectrophotometer 6, 12, 24 and 48 h after the transfection of recombinant vector into the TM4 cell line. Results The entire coding sequence of Cox7a2 was cloned with 252 bp length. Plasmid pEYFP-C1-Cox7a2 vector was constructed and the positive clones were verified by restriction enzymes digestion and DNA sequencing. The transfection efficiency of the TM4 cell line was about 70% and 37000 D fusion protein was obtained. The COX activities were (0.642±0.051), (0.542±0.049), (0.311±0.021) and (0.216±0.010) U/mg 6, 12, 24 and 48 h after the transfection of recombinant vector in the TM4 cell line. Meanwhile, the COX activities were (0.714±0.064) and (0.653±0.031) U/mg in non-tranfected and naked vector group respectively. Compared with the non-tranfected group, COX activity decreased significantly 12, 24 and 48 h after the transfection. Conclusions The recombint plasmid vector was successfully constructed. Cox7a2 gene has an inhibiting effect on COX activity and may play an important role in the regulation of COX activity in mouse Sertoli cell line TM4.

Objective To evaluate the expression of human vascular endothelial cell growth factors 165 (hVEGF165) gene transfected into fibroblasts by recombinant adenovirus and study the repairing effect of this cells on radiated skin ulcer in rats.Methods The recombinant adenovirus with hVEGF165 was established and transfected to rat primary fibroblasts, and its expression of hVEGF165 in fibroblasts was identified with real-time PCR, immunocytochemistry and Western blot.Twenty four clean grade SD rats of were irradiated locally with 50 Gy γ rays to generate an animal model of radiation skin injury.The hVEGF165-transfected cells were injected to the irradiated site under rat skin 7 d post-irradiation.The therapeutic effects on the irradiated skin wound were evaluated through general observation as well as histological staining of HE.The expression of hVEGF in the irradiated skin tissue with fibroblasts injection was analyzed by Real-time PCR.Results The hVEGF165 gene was overexpressed in the transfected cells and approached to 88 373-fold bigger compared to controls transfected with blank vectors, and an extensive expression of VEGF in the cytoplasm of transfected cells was observed by immunohistochemistry.VEGF protein with the relative molecular mass of 23 000 was also detected in cell lysate by Western blot.The local skin ulcers in rats occurred about two weeks after irradiation.In the hVEGF165-transfected group, the average area of radiation-injured skin was 40.2 mm2, about 57％ less than that of the control group transfected with blank vectors so that the healing time was shorten by 6 days.The relative concentration of hVEGF mRNA in the skin tissue of rats injected with hVEGF165-transfected cells were 5.15-fold and 4.15-fold bigger compared to that of controls (t =3.385,3.220, P ＜ 0.05) at 3 and 7 d after administration.Conclusions The primary fibroblasts transfected with hVEGF165 gene could efficiently release VEGF to the irradiated skin tissue and promote the recovery of irradiation skin ulcers by shortening the healing time and thus enhanced the therapeutic effect on skin wounds.

Objective To evaluate pathogens and antimicrobial resistance of pathogens causing refractory pneumonia in children.Methods Children with refractory pneumonia who admitted to a hospital between May 2008 and December 2014 were performed bronchoscopy,and bronchoalveolar lavage fluid (BALF)were performed bacterial culture and antimicrobial resistance testing.Results 1 693 patients were recruited in the study,273 bacterial isolates were isolated from BALF speci-mens of 226 children,gram-positive bacteria accounted for 38.10% (104/273 ),the main gram-positive bacteria were Streptococcus pneumoniae (n=71)and Staphylococcus aureus (n=23);gram-negative bacteria accounted for 58.24%(159/273),including 44 isolates of Haemophilus parainfluenzae ,28 Klebsiella pneumoniae ,19 Escherichia coli ,and 17 Pseud-omonas aeruginosa ;10 isolates of fungi were also detected,8 of which were Candida albicans .The sensitivity of Streptococ-cus pneumoniae to quinolones,ceftriaxone and cefotaxime were high.Methicillin-resistant Staphylococcus aureus (MRSA) positive rate was 26.32%.ESBLs-producing rate of Haemophilus parainfluenzae and Klebsiella pneumoniae was 32.72% and 62.96% respectively.Conclusion The major pathogens causing refractory pneumonia were Streptococcus pneumoniae and Haemophilus parainfluenzae ,empirical treatment should be conducted accordingly,antimicrobial resist-ance should be considered if therapeutic effect is poor,and targeted therapy should be performed according to cultured re-sults and antimicrobial susceptibility testing result.

Objective To analyze the differences in gene expression between human lung giant cell carcinoma cell strains of high metastatic 95D and low metastatic 95C and to screen lung cancer metastasis-associated genes using cDNA microarray.Methods The mRNAs were extracted from human lung giant cell carcinoma cell strains of high metastatic 95D and low metastatic 95C,and reversely transcribed to the cDNAs and labeled by Cy5-dUTP and Cy3-dUTP to prepare the hybridization probes.The mixed probes were hybridized to the cDNA microarray chip.The information was obtained by managing the cDNA microarray chip with Scan Array 3000 scanner and ImaGene3.0 software.A part of genes whose expressions changed in cDNA microarray analysis were further identified by RT-PCR.Results The cDNA microarray analysis showed that the expressions of 466 genes changed,among which 108 pairs of Double Gene had the same GenBank ID and some of the genes,including Fln29,RUVBL2,C14orf3 et al,were further confirmed by RT-PCR.The results of RT-PCR was coincided well with the cDNA microarray results.Conclusion Many different genes are involved in the metastasis of lung cancer.cDNA microarray technique might be a useful method in screening lung cancer metastasis-associated genes.

Objective:To evaluate the value and identify the prognosic factors of postoperative radiotherapy (PORT) in completely resected stage Ⅲ(pN 2) lung adenocarcinoma patients with epidermal growth factor receptor (EGFR) wild-type who received adjuvant chemotherapy. Methods:Clinical data of 172 patients with stage Ⅲ(pN 2) EGFR wild-type lung adenocarcinoma who underwent radical resection and adjuvant chemotherapy from 2009 to 2016 were retrospectively analyzed. All patients received platinum-based adjuvant chemotherapy combining two drugs for>4 cycles, and divided into the PORT group and the non-PORT group. The survival rate was calculated by Kaplan- Meier method and log-rank test, and multivariate prognostic analysis was performed by Cox’s regression model. Results:Among 172 patients, the median overall survival (OS), 3-year and 5-year OS rates were 40 months, 55.9% and 28.3%, respectively. The median disease-free survival (DFS), 3-year and 5-year DFS rates were 17 months, 24.5% and 13.0%, respectively. DFS was significantly improved in the PORT group (29 months vs. 13 months, P=0.001), whereas OS did not significantly differ between two groups (51 months vs. 38 months, P=0.151). In subgroup analysis, DFS of patients with multistation N 2 or the number of N 2 metastases of≥3 or skip N 2 in the PORT group was significantly longer ( P<0.05), whereas PORT exerted no significant effect on OS ( P>0.05). Conclusions:For patients with completely resected stage Ⅲ(N 2) EGFR wild-type lung adenocarcinoma receiving adjuvant chemotherapy, PORT might increase DFS and have a trend toward longer OS. However, these findings remain to be validated by large sample size investigations.

Background: Metabolic diseases pose considerable burden on the healthcare system worldwide, indi-cating the significance of prevention and treatment. In constitution theory of traditional Chinese med-icine, phlegm-dampness constitution (PDC) is the common basis of metabolic diseases. In clinical practice, Huatan Qushi (HTQS) decoction targeting on PDC can effectively improve metabolic indicators. However, its underlying biochemical mechanism still remains unclear.Methods: Eight PDC participants received HTQS decoction for three months. Their blood was collected at baseline and 1 and 3 months after intervention started. Related biomedical indicators were detected. High-throughput sequencing and RT-qPCR were used for validation. Due to the missing data, repeated measures with missing values in mixed models were used. Results: After 3-month treatment, HDL-C level increased (P<.001) and FBG, FINS, and HbA1c all showed decreasing trend at different time points (all P < .05). After miRNA high-throughput sequencing, compared with the baseline, differential miRNAs at 1 and 3 months were screened, and target gene prediction and KEGG pathway enrichment analysis were performed. The results displayed that metabolic disease-related pathways mainly included pathways in cancer, PI3K-Akt signaling pathway, etc. Further, RT-qPCR showed that hsa-miR-1237-3p differed statistically (P =.008). Then we validated the target genes of hsa-miR-1237-3p in the"Pathways in Cancer"pathway including SDF1, AC, CRK, and HGF, also known as upstream target genes of PI3K/AKT pathway. The results showed that two indicators of CRK and HGF were in statistical significance (P=.045 and P=.036, respectively). Conclusion: PDC serves as a common basis for various metabolic diseases. Through adjusting PDC, HTQS decoction can improve biomedical indicators including blood glucose, HbA1c, insulin, and HDL-C. The target pathway is"Pathways in cancer". Specifically, HTQS decoction acts on targets of CRK and HGF by regulating hsa-miR-1237-3p, and probably exerts effects on their downstream PI3K/AKT pathway.

Background::Heterogeneity of tumor cells and the tumor microenvironment (TME) is significantly associated with clinical outcomes and treatment responses in patients with urothelial carcinoma (UC). Comprehensive profiling of the cellular diversity and interactions between malignant cells and TME may clarify the mechanisms underlying UC progression and guide the development of novel therapies. This study aimed to extend our understanding of intra-tumoral heterogeneity and the immunosuppressive TME in UC and provide basic support for the development of novel UC therapies.Methods::Seven patients with UC were included who underwent curative surgery at our hospital between July 2020 and October 2020. We performed single-cell RNA sequencing (scRNA-seq) analysis in seven tumors with six matched adjacent normal tissues and integrated the results with two public scRNA-seq datasets. The functional properties and intercellular interactions between single cells were characterized, and the results were validated using multiplex immunofluorescence staining, flow cytometry, and bulk transcriptomic datasets. All statistical analyses were performed using the R package with two-sided tests. Wilcoxon-rank test, log-rank test, one-way analysis of variance test, and Pearson correlation analysis were used properly.Results::Unsupervised t-distributed stochastic neighbor embedding clustering analysis identified ten main cellular subclusters in urothelial tissues. Of them, seven urothelial subtypes were noted, and malignant urothelial cells were characterized with enhanced cellular proliferation and reduced immunogenicity. CD8 + T cell subclusters exhibited enhanced cellular cytotoxicity activities along with increased exhaustion signature in UC tissues, and the recruitment of CD4 + T regulatory cells was also increased in tumor tissues. Regarding myeloid cells, coordinated reprogramming of infiltrated neutrophils, M2-type polarized macrophages, and LAMP3 + dendritic cells contribute to immunosuppressive TME in UC tissues. Tumor tissues demonstrated enhanced angiogenesis mediated by KDR + endothelial cells and RGS5 +/ACTA2 + pericytes. Through deconvolution analysis, we identified multiple cellular subtypes may influence the programmed death-ligand 1 (PD-L1) immunotherapy response in patients with UC. Conclusion::Our scRNA-seq analysis clarified intra-tumoral heterogeneity and delineated the pro-tumoral and immunosuppressive microenvironment in UC tissues, which may provide novel therapeutic targets.

Objective: To investigate the clinical manifestations, pathological features, and treatment of oral and maxillofacial pyogenic granulomas induced by camrelizumab. Methods: A case of pyogenic granuloma of the gums and lips caused by camrelizumab was reported along with a literature review. Results: After 4 months of treatment with camrelizumab for liver cancer, the patient developed systemic reactive capillary hyperplasia (RCH), followed by multiple masses on the lower lip and gingiva. After periodontal therapy, the masses on the lower lip and the gingiva were removed, and camrelizumab administration was stopped. The pathological result was gingival pyogenic granuloma/granulomatous hemangioma. No new masses were found in the oral cavity during postoperative follow-up. A review of the literature showed that RCH is the most common adverse drug reaction to camrelizumab but it occurs infrequently in the oral cavity. At present, the etiology of RCH has not been clarified, but the research has shown that camrelizumab may trigger tissue proliferation into hemangiomas by activating vascular endothelial cells, and the combined use of camrelizumab is safer than single use. RCH is self-limiting and most cases resolve spontaneously after discontinuation of the drug. If the mass causes dysfunction, surgical excision is feasible. Conclusion Camrelizumab can cause oral and maxillofacial reactive capillary hyperplasia complicated by pyogenic granuloma.