Purpose:To observe the curative effect and the clinical-benefit-response of patients with advanced pancreatic cancer who used a combine of Gemcitabine,5-FU and PDD.Methods:The treatment with Gemcitabine (800 mg/m 2) was followed by 5-FU(600 mg/m 2) and DDP (30 mg/m 2),administered on days 1,8 and 15 of 28-day schedule. The curative effect was estimated according to the standard of WHO and the clinical benefit response was evaluated at the same time. The standard of CBR included pain,physical states and body weight. Results:Twenty-five (25/29) patients received over two cycles chemotherapy. 6(24%) partial response (PR), 12 (48%) stable disease and 7(28%) progressive disease were observed. According to the standard of CBR, 18(62.1%) patients achieved the clinical-benefit-response level. Therapy was well tolerated. The main toxicities included that the alimentary tract reactions and the marrow depression. Grade 3-4 thrombocytopenia was reported in 8 (27.6%) patients. Conclusions:The combination of GEM, 5-FU and DDP is a moderatedly active treatment with tolerable side-effects. It has definite response rate and high CBR rate in pancreatic cancer.

Objective:To explore the immunosuppression effects and tocolysis of fructus polygoni orientalis on abortion mice induced by LPS.Methods:Mice of Kunming (55 mice) were divided into control group (group A ,10 mice) and experimental group. Experimental group were divided into group B ( intraperitoneal injection LPS ) , group C ( ingestion fructus polygoni orientalis ) , and group D (ingestion fructus polygoni orientalis and intraperitoneal injection LPS ),each group of 15 mice.Then the pregnancy results were observed ,the positive of α-NAE+and the varity of CD 14+and CD204+macrophages ,TNF-αin the uteri were identified by enzyme-histochemistry ,immunohistochemistry and ELISA.Results:The abortion rate and the embryo resorbing rate were all 100%( P<0.01 ) in group B.But there was the decreased abortion rate of 13.33% in group D .The embryo resorbing rate decreased to 10.39%.The number and positive cell area of α-NAE+and CD14+macrophages in the uteri of gestation mice of group B was greatly increased comparing with group A ( P<0.01 ) .These effects outside of myometrium of group D were remarkably increased comparing with group A (P<0.01),but there was no distinct difference in the inside of myometrium and function layer.The number and positive cell area of CD204+macrophages in group C and D was greatly increased comparing with group A and B ( P<0.01 ) .The TNF-αcontents in the uteri of mice in group B were greatly increased comparing with group A (P<0.01),but the positive cell area of CD14,CD204 were close to normal levels in group D.Conclusion:The effect of miscarriage induced by LPS is antagonized by fructus polygoni orientalis through inhibiting the phenotype ,activity and function characteristics of macrophages in the uteri of gravidity mice.

Objective:To explore the adjustment factors to the migration and functional activity on macrophages in the uterine of inflammatory mice induced by LPS.Methods:150 Kunming female mouse were divided into control group (group A),LPS model group (group B),MCP-1 blocking-up group (group C),the mice uterines were extracted separately at hour 1,3,6,12,24.The number of CD14+ macrophages and the expression of CD14 macrophages were detected by Immunohistochemistry,ELISA detects the expression of TNF-α and MCP-1.Results:①Compared with group A,the number of CD14+ macrophages and the expression of CD14 in endometrium,myometrium,perimrtrium of group B were highly significantly increased (P<0.01) at every time points,the endometrium,myometrium of group C were closely to normal level at 1,3,6 h;compared with group B,the number of CD14+ macrophages and the expression of CD14 were highly significantly decreased(P<0.01)at 1,3,6 h of perimrtrium of group C and every time points of endometrium,myometrium of group C.②Compared with group A,the content of TNF-α and MCP-1 were highly significantly increased (P<0.01) at every time points of group B and 12,24 h of group C;compared with group B,the content of TNF-α and MCP-1 of group C were highly significantly decreased(P<0.01) at every time points.Conclusion:The migration of macrophages and the expression of CD14 and TNF-α in the uterine of inflammatory mice induced by LPS were regulated by MCP-1.

Objective To establish a Juema minipig model of myocardial infarction, to evaluate the clinical indi?ces in the model pigs, and to explore the relationship between gene expression and metabolic decompensation. Methods 13 male Juema minipigs were randomly divided into control (Sham, n=5), myocardial infarction (MI, n=5) and normal control (for evaluating the recovery condition after surgery, n=3) groups. In the MI group, the ligation was done at the left descending coronary artery around the 1/3 distance to heart apex. Four weeks after the surgery, the cardiac function and serum biochemistry was analyzed. The histological changes and gene expression profiles in the myocardium in the peri?infarct area were exanimated. Results Ultrasonic images showed that the infarction was formed, the ejection fraction and fraction shortening were significantly reduced in the MI group ( ~32% and ~40% less than those of the sham group). Histological examination showed that myocardial fibers at the peri?infarct area were broken, dissolved, and there was con?nective tissue hyperplasia with increased neutrophil and lymphocyte infiltration. Microarray analysis revealed that two myo?cardial remodeling and pathology mediating pathways, three inflammation?related pathways, and 8 metabolic pathways ( in?cluding fatty acid, amino acid, and glucose metabolic pathways) were significantly changed. Conclusions We have suc?cessfully established a Juema minipig model of myocardial infarction. The less branches of the left descending coronary ar?tery allow us to establish a stable model by surgery with comparable characteristics in the clinic indices. The results of this study provides useful reference characteristics of an animal model with characteristic changes in the peri?infarct area.

AIM: To analyze the influencing factors of postoperative dry eye complication after corneal transplantation and to build a nomogram prediction model.METHODS: Clinical data were collected on 117 patients who underwent corneal transplantation at our hospital from March 2021 to February 2023. They were divided into dry eye group(n=96)and non-dry eye group(n=21)according to whether there was a postoperative dry eye. The risk factors of postoperative complication of dry eye after corneal transplantation were analyzed, the nomogram prediction model for predicting postoperative complication of dry eye after corneal transplantation was constructed, and the internal validation of the model and the prediction efficacy were assessed by calibration curves and decision curves, respectively.RESULTS: Comorbid diabetes mellitus, comorbid sleep disorders, comorbid meibomian gland dysfunction, chronic eye drop abuse, chronic corneal contact lens wear, interleukin-6(IL-6), and tumor necrosis factor-α(TNF-α)were the risk factors for the complication of dry eye after corneal transplantation(P<0.05). The nomogram model predicted a C-index of 0.890(95% CI 0.877-0.903). The nomogram model had a threshold >0.07, and the nomogram model provided higher net clinical benefit than the single index in all cases.CONCLUSION: The nomogram model built in this study based on the factors affecting the complication of dry eye after corneal transplantation has a good predictive value for the complication of dry eye after corneal transplantation.

Objective To explore the effect ofpolycyclic aromatic hydrocarbons (PAHs) and p16,FHIT gene CpG island methylation,as well as their interaction in cervical intraepithelial neoplasias.Methods Objects of this study were from a cohort of cervical lesions study in Yangqu county of Shanxi province.All the patients were diagnosed pathologically,that including 83 patients with high-grade cervical intraepithelial neoplasia (CIN Ⅱ/Ⅲ),86 patients with low-grade cervical intraepithelial neoplasia (C1N Ⅰ) and another 91 women under normal cervical (NC) condition.1-hydroxy pyrene in the urine was detected by high performance liquid chromatography (HPLC)while CpG island methylation status of tumor suppressor gene p16 and FHIT were measured by methylation-specifc polymerase chain reaction (MSP).Data were analyzed with Kruskal-Wallis H test,chi-square test and trend of chi-square test.Logistic regression models were used to estimate the odds ratio (OR) and corresponding 95％ confidence intervals (95％CI) between influencing factors and the cervical disease by using the SPSS statistical software (version 20.0).The interaction under study was evaluated by using the generalized multifactor dimensionality reduction (GMDR) model.Results Level of 1-hydroxy pyrene (H=50.743,P＜0.001) and the high exposure rate of 1-hydroxy pyrene (trend x2=20.146,P＜0.001) were gradually increasing along with the severity of cervical intraepithelial neoplasia.The CpG island methylation rates ofpl6,FHIT in CIN] and CIN Ⅱ/Ⅲl group were higher than that in NC group,and gradually increasing along with the severity of cervical intraepithelial neoplasia (trend x2=9.75,P=0.002;trend x 2 =10.39,P=0.001).Results from the GMDR model showed that interaction existed among the high exposure of l-hydroxy pyrene and the CpG island methylation ofpl6,FHIT in CIN Ⅰ and CIN Ⅱ]/Ⅲ group.Conclusion Under the high exposure of 1-hydroxy pyrene and the CpG island methylation of p16,FHIT appeared to have increased the risk of cervical intraepithelial neoplasia and causing synergistic effect in cervical intraepithelial neoplasia.

Objective To explore the effect of Src on cervical cancer cells through ERK signal transduction pathway.Methods Experimental study was carried out in vitro.Cervical cancer cell lines Hela (HPV-positive) and C33A (HPV-negative) were treated with Src kinase inhibitor PP2.Then,the cell cycle and apoptosis of each group were evaluated by using flow cytometry (FCM).Western blotting and Real-time PCR were used to detect the levels of the expression of ERK 1/2,c-Fos and c-Jun mRNA and protein respectively.The database was established and analyzed with SPSS statistical software (version 20.0).Results After down-regulating Src,the cell proliferation was inhibited and cell apoptosis was induced.The proportions of G0/G 1 stage of Hela and C33A cell in cell cycle increased while G2/M and S stages decreased.Meanwhile,the mRNA levels of ERK 1,ERK 2,c-Fos and c-Jun increased.And the expression levels of ERK 1/2,phosphorylated ERK 1/2 (p-ERK 1/2)and phosphorylated c-Fos (p-c-Fos) protein decreased,while c-Jun and phosphorylated c-Jun (p-c-Jun)protein expression increased.In addtion,the change level of Hela cell,p-ERK 1/2 and c-Fos protein were lower than that of C33A cell before and after the Src inhibition.Conclusions Src,involved in regulating the expression of key factors of the ERK signal transduction pathway including p-ERK 1/2 and p-c-Fos,might be capable of promoting the proliferation of cervical cancer cells and inhibiting their apoptosis.The infection with HPV might have adjustable effect on this process.

Clustered regulatory interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 nuclease (Cas9), the third-generation genome editing tool, has been favored because of its high efficiency and clear system composition. In this technology, the introduced double-strand breaks (DSBs) are mainly repaired by non-homologous end joining (NHEJ) or homology-directed repair (HDR) pathways. The high-fidelity HDR pathway is used for genome modification, which can introduce artificially controllable insertions, deletions, or substitutions carried by the donor templates. Although high-level knock-out can be easily achieved by NHEJ, accurate HDR-mediated knock-in remains a technical challenge. In most circumstances, although both alleles are broken by endonucleases, only one can be repaired by HDR, and the other one is usually recombined by NHEJ. For gene function studies or disease model establishment, biallelic editing to generate homozygous cell lines and homozygotes is needed to ensure consistent phenotypes. Thus, there is an urgent need for an efficient biallelic editing system. Here, we developed three pairs of integrated selection systems, where each of the two selection cassettes contained one drug-screening gene and one fluorescent marker. Flanked by homologous arms containing the mutated sequences, the selection cassettes were integrated into the target site, mediated by CRISPR/Cas9-induced HDR. Positively targeted cell clones were massively enriched by fluorescent microscopy after screening for drug resistance. We tested this novel method on the amyloid precursor protein (APP) and presenilin 1 (PSEN1) loci and demonstrated up to 82.0% biallelic editing efficiency after optimization. Our results indicate that this strategy can provide a new efficient approach for biallelic editing and lay a foundation for establishment of an easier and more efficient disease model.
Alleles ; CRISPR-Cas Systems ; DNA End-Joining Repair ; Gene Editing/methods* ; Recombinational DNA Repair

Objective:To summarize and analyze the clinical data of Chinese patients with colony-stimulating factor 1 receptor (CSF1R)-related leukoencephalopathy, and clarify the phenotypic and genetic characteristics of Chinese patients.Methods:Medical history of patients with CSF1R-related leukoencephalopathy diagnosed from April 1, 2018 to January 31, 2021 in the department of neurology of 22 hospitals in China was collected, and scores of Mini-Mental State Examination (MMSE), Montreal Cognitive Assessment Scale (MoCA), magnetic resonance severity scale were evaluated. Group comparison was performed between male and female patients.Results:A total of 62 patients were included, and the male-female ratio was 1∶1.95. The age of onset was (40.35±8.42) years. Cognitive impairment (82.3%, 51/62) and motor symptoms (77.4%,48/62) were the most common symptoms. The MMSE and MoCA scores were 18.79±7.16 and 13.96±7.23, respectively, and the scores of two scales in male patients (22.06±5.31 and 18.08±5.60) were significantly higher than those in females (15.53±7.41 , t=2.954, P=0.006; 10.15±6.26, t=3.328 , P=0.003). The most common radiographic feature was bilateral asymmetric white matter changes (100.0%), and the magnetic resonance imaging severity scale score was 27.42±11.40, while the white matter lesion score of females (22.94±8.39) was significantly higher than that of males (17.62±8.74 , t=-2.221, P<0.05). A total of 36 CSF1R gene mutations were found in this study, among which c.2381T>C/p.I794T was the hotspot mutation that carried by 17.9% (10/56) of the probands. Conclusions:The core phenotypic characteristics of CSF1R-related leukoencephalopathy in China are progressive motor and cognitive impairment, with bilateral asymmetrical white matter changes. In addition, there exist gender differences clinically, with severer cognitive impairment and imaging changes in female patients. Thirty-six CSF1R gene mutations were found in this study, and c.2381T>C/p. I794T was the hotspot mutation.

Arsenic trioxide (ATO) has been identified as an effective treatment for acute promyelocytic leukemia (APL) but is much less effective against solid tumors such as hepatocellular carcinoma (HCC). In the search for ways to enhance its therapeutic efficacy against solid tumors, we have examined its use in combination with a novel derivative of β-elemene, N-(β-elemene-13-yl)tryptophan methyl ester (ETME). Here we report the effects of the combination on cell viability, apoptosis, the cell cycle and mitochondria membrane potential (MMP) in HCC SMMC-7721 cells. We found that the two compounds acted synergistically to enhance antiproliferative activity and apoptosis. The combination also decreased the MMP, down-regulated Bcl-2 and pro-proteins of the caspase family, and up-regulated Bax and BID, all of which were reversed by the p53 inhibitor, pifithrin-α. In addition, the combination induced cell cycle arrest at the G2/M phase and reduced tumor volume and weight in an xenograft model of nude mice. Overall, the results suggest that ETME in combination with ATO may be useful in the treatment of HCC patients particularly those unresponsive to ATO alone.