Objective To explore the effect of new hardener gathered cinnamyl alcohol (hardening of the oxygen ethyl alcohol)and anhydrous ethanol in the treatment of endometriosis cysts guided by ultrasound.Methods Two groups of cases,a group of 36 cases of endometriosis cyst in anhydrous ethanol sclerotherapy under the guidance of ultrasound(anhydrous ethanol group),another group of 30 cases of endometriosis cyst under the ultrasonic guided into 1% poly cinnamyl alcohol hardening treatment(poly cinnamyl alcohol group).Results Following -up every 3 month,the longest time was 2 years follow -up,the shortest time was 9 months.The anhydrous ethanol group:the to-tal effective rate was 95.0% (excluding a patients discontinue treatment due to severe pain),and the recurrence rate of 12.5% in the first year,second year recurrence rate 17.5%;The poly cinnamyl alcohol group:the total effective rate was 94.2%,the recurrence rate of in the first year was 11.7%,second year recurrence rate was 17.6%.Conclusion Anhydrous ethanol and cinnamyl alcohol hardening treatment of endometriosis cyst accuracy guided by ultrasound is high,the treatment effect,the recurrence rate has no obvious difference,but small poly cinnamyl alcohol irritating to human body,the patient has no obvious pain,easy to accept,and people feel ethanol treatment most patients with pain,with occasional patients discontinue treatment due to severe pain.

Objective: To evaluate the diagnostic significance of CEA specific antibodies in the serum of patients with gastrointestinal malignancies. Methods: The CR4 levels were tested by an.The circulating antibodies against CEA were tested by an enzyme-linked immunoassay (ELISA) .The specificity of the CEA antibodies was carried out by an inhibition assay. Results: The CEA was elevated( ≥15 ng/ml) in 21 patients with gastrointestinal malignancies(30.9%) and in 1 patient with benign gastrointestinal conditions( hyperplastic polys, ulcers, pancreatitis, etc)(3.3%) .Anti-CEA IgG antibodies were detected in 24 patients with gastrointestinal malignancies(35.3%) and in 1 health individual (2.5%).Conduskm: In this study ,a high number of gastrointestinal malignancies patients have circulating CEA specific antibodies in their sera and these antibodies may be used as diagnostic markers.

Objective To observe the prophylactic effects of a HSV-2gD2DNA vaccine in guinea pigs challenged with HSV-2strains.Methods Female guinea pigs were divided into3groups with10each,which was immunized intramuscularly with100?g of pc-gD plasmids(recombinant HSV-2DNA vac-cine),or with pcDNA3blank plasmids,with normal saline as control,respectively.Two booster injections were given on day7and day21.Sera were collected for virus neutralization test on day0,day28,and day56.The animals were challenged with HSV-2strain sav intravaginally,and lesions induced on the external genital skin were scored between day1and day21after challenge.Results The titer of neutralizing anti-body to HSV-2was much higher in the sera from animals immunized by pc-gD plasmids than that from ani-mals immunized by pcDNA3blank plasmids or normal saline.Furthermore,the lesion scores on external genital skin were significantly decreased in pc-gD group than those in other two groups with either primary or recurrent infections.Conclusion The constructed gD2vaccine can efficiently protect guinea pigs from genital infection and reduce recurrent infection induced by latent herpes simplex virus.

Xihuang pill,a famous anticancer traditional Chinese medicine formula,has curative effect in clinical application. It can inhibit the growth and invasion of tumor cells and cancer stem cells,prevent angiogenesis,and reverse the tumor immunosup-pressive microenvironment. This review summarizes the reported Xihuang pill' s anticancer mechanism and clinical application on colorectal cancer,breast cancer,liver cancer,so as to provide a reference for the further research and development on anticancer ap-plication of Xihuang pill.

Objective To explore the preparation of recombinant human cystatin C (Cys C ) and its polyclonal antibody . Methods The gene sequences encoding Cys C were designed according to characteristics of E .coli protein-coding ,and the target gene was artificially synthesized and expressed in E .coli .The target protein was purified by affinity chromatography and was used to immunize the New Zealand white rabbits .Western blot and enzyme-linked immunosorbent assay(ELISA) were adopted to detect the recombinant human Cys C and its polyclonal antibodies in the rabbits serum .Results Purity of prepared recombinant human Cys C was more than 90% .Anti-recombinant human Cys C antibody showed good binding activities and specificity .Conclusion Prepared recombinant human Cys C and its antibody can be used in further experimental research and clinical application .

Objective:To investigate the correlation between variations in mitochondrial DNA (mtDNA) control region (D-loop) and keloids.Methods:A total of 216 patients with keloids were collected from Department of Dermatology, the First Affiliated Hospital of Kunming Medical University from 2016 to 2019. Total DNA was extracted from peripheral blood samples of all the patients, as well as keloid tissues and perilesional normal skin tissues of 25 patients with keloids. Peripheral blood samples were collected from 299 health checkup examinees without keloids in Health Examination Center, the Affiliated Hospital of Yunnan University, who served as controls. PCR amplification and Sanger sequencing were performed on the mtDNA D-loop region, and mutation sites in each sample were analyzed by comparisons with the revised Cambridge Reference Sequence (rCRS) . Haplogroups were assigned in the 2 groups by using Phylotree-mtDNA tree Build 17. Mutations in the mtDNA D-loop region were compared among keloid tissues, perilesional normal skin tissues and peripheral blood samples. A median-joining network was constructed via network 5.0 software. Binary logistic regression analysis was performed to investigate the correlation between haplogroup frequencies and the occurrence of keloids, and chi-square, t and t′ tests were used to analyze clinical data. Results:Among the 216 patients with keloids, variations in mtDNA D-loop region were classified into 10 haplogroups, including A, B, D, R9, G, M*, M7, M8, M9 and N9, with the haplogroups R9 and M9 showing the highest (21.3%, 46/216) and lowest (0.9%, 2/216) frequencies respectively. The frequencies of haplogroups M7 ( P=0.040, OR=0.248, 95% CI: 0.066 - 0.937) and N9 ( P=0.048, OR=0.191, 95% CI: 0.037-0.986) were significantly lower in the patients with keloids than in the controls. The median-joining network plot showed that the distribution pattern of the haplogroup M7 differed between the patients with keloids and controls. Significantly less number of lesional sites and younger age of onset were observed in the patients with haplogroup M7 compared with those with non-M7 haplogroups ( P=0.000 1, 0.045, respectively) . Conclusion:The haplogroup M7 is correlated with the occurrence of keloids, and may be a potential protective factor for keloid formation.

OBJECTIVETo screen for the inhibitor of vascular endothelial growth factor (VEGF) 165 from random peptide library.

METHODSPositive phage clones were rescued after two rounds of panning and competitive elution. Its affinity activity to KDR was monitored through ELISA, immunohistochemical method, Chicken CAM assay and MTT.

RESULTSFive specific binding positive target molecule phage clones were obtained which were able to bind to cells whose surface had high KDR, among which, clone 3 and 13 could effectively block the vascularization of the chorioallantoic membrane of chick embryo, but they were not inhibitive on the proliferation of high KDR expression cells.

CONCLUSIONThe peptides, being the inhibitors of VEGF, may be useful in the treatment of cancers.

Animals ; Binding Sites ; Endothelial Growth Factors ; antagonists & inhibitors ; metabolism ; Enzyme-Linked Immunosorbent Assay ; Humans ; Intercellular Signaling Peptides and Proteins ; metabolism ; Lymphokines ; antagonists & inhibitors ; metabolism ; Peptide Library ; Peptides ; pharmacology ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

Objective:To investigate whether IL-7-secreting oncolytic herpes simplex virus(HSV)could activate CD8+T cells and inhibit the growth of hepatocellular carcinoma.Methods:The expression of IL-7 was detected by Western blot.The in vitro cleavage of tumor cells by tumor oncolytic virus HSV and HSV-IL-7 were detected by crystal violet staining.The tumor inhibition ability of HSV-IL-7 and HSV were detected in subcutaneous transplanted tumor model.Levels of IL-7,IFN-γ and TNF-α in serum and tumor tissues were determined by ELISA.The infiltration of CD8+T cells in tumor tissues was detected by immunohistochemistry.Flow cytometry was used to detect Granzyme B secretion in CD8+T cells infiltrated by tumor.Results:Tumor cells infected with HSV-IL-7 expressed high level of IL-7.Both HSV and HSV-IL-7 can effectively lyse B16-F10,CT-26 and H22 tumor cell lines in a dose-dependent manner in vitro.HSV-IL-7 could significantly inhibit the growth of H22 hepatoma cells in vivo(P<0.01)and prolong the survival time of tumor-bearing mice(P<0.001).HSV-IL-7 could significantly increase the IL-7 content in tumor sites(P<0.000 1),and effectively increase the number of tumor infiltrating CD8+T cells(P<0.001).HSV-IL-7 significantly enhanced Granzyme B secretion of tumor-infiltrating CD8+T cells and IFN-γ and TNF-α in tumor tissues(P<0.000 1).Conclusion:HSV-IL-7 has well tumor inhibition activity in vivo and in vitro.It also can activate the anti-tumor activity of CD8+T cells in vivo by secreting IL-7,inhibit tumor growth and prolong the survival time of tumor-bearing mice.

Asymptomatic spleen-stomach diseases refer to diseases without related symptoms and signs of abdo-minal pain， bloating， diarrhea an others in patients， but showing lesions or pathological changes discovered by modern medical techniques such as endoscopy， CT， MRI. The four examination techniques of traditional Chinese medicine （TCM） are based on symptoms and signs of patients， which are the advantage of TCM but also have certain limitations. In the context of the increasingly modernized diagnosis and treatment in TCM， it is proposed to expand the application of the four examination techniques from three aspects including microcosmic syndrome differentiation， data sharing， and artificial intelligence in asymptomatic spleen-stomach diseases， in order to achieve the goals of dynamically observing the disease process， collecting disease data in multiple dimensions， and intelligently processing disease data. This will strengthen the modern requirements of early diagnosis and treatment in TCM， and highlight the advantages of TCM in “treating disease before it arises and treating the symptoms beforehand”.

Objective : To investigate the regulatory role of ubiquitin-specific protease 10 ( USP10) on the protein expression of Krüppel-like factor 4 (KLF4) and its impact on the proliferation and invasion ability of hepatocellular carcinoma (HCC) cells． Methods : The protein expression differences of USP10 and KLF4 in normal liver cell line L02 and HCC cell lines，including HepG2，HUH7，HCCLM3 were detected by immunoblotting ( Western blot ) methods．HCCLM3 and HUH7 cells were selected，and lentiviral particles overexpressing or silencing USP10 ( oe- USP10 or sh-USP10) was transfected into the cells，and they were designated as the oe-USP10 group and oe-NC group，respectively．Immunoprecipitation ( Co-IP) experiments were conducted to examine whether USP10 could di- rectly interact with KLF4 in HCCLM3 or HUH7 cells．The Co-IP assay was repeated in HCC cells transfected with oe-USP10 or sh-USP10，with the addition of the proteasome inhibitor MG132，which used to detect the ubiquitina- tion level of KLF4 protein in the transfected HCC cells．The pcDNA3. 1 vector containing overexpressed KLF4 or its negative control plasmid (pc-KLF4 or pc-NC) was co-transfected into cells of the sh-USP10 group or sh-NC group. These cells were designated as the sh-NC + pc-NC group，sh-USP10 + pc-NC group，sh-NC + pc-KLF4 group，and sh-USP10 + pc-KLF4 group．The cell proliferation activity of each group was measured using the CCK-8 assay，and the cell invasion ability was assessed using the Transwell assay. Results : Compared to L02 cells，the protein expres- sion of USP10 and KLF4 significantly decreased in HepG2，HUH7，HCCLM3，and other cells (P＜0. 05) ．In HC- CLM3 and HUH7 cells，USP10 protein directly interacted with KLF4．Furthermore，treatment with MG132 resulted in a time-dependent increase in KLF4 protein expression in HCCLM3 and HUH7 cells．Silencing USP10 increased the ubiquitination of KLF4 in HCCLM3 or HUH7 cells，while overexpressing USP10 decreased the ubiquitination level of KLF4 in cells．Compared to the sh-NC + pc-NC group，both the proliferation activity and invasion ability of HCCLM3 and HUH7 cells significantly increased in the sh-USP10 + pc-NC group (P ＜0. 01) ，while they signifi- cantly decreased in the sh-NC + pc-KLF4 group and sh-USP10 + pc-KLF4 group (P＜0. 05) ．Compared to the sh- USP10 + pc-NC group，the proliferation activity and invasion ability of cells significantly decreased in the sh-USP10 + pc-KLF4 group (P＜0. 05) ． Conclusion USP10 can promote the stability of KLF4 protein through deubiquiti- nation in HCC cell lines，thereby inhibiting the proliferation and invasion of tumor cells.