Abstract: Objective　The method of "molecular docking - molecular dynamics - in vitro pharmacology" was used to screen and optimize small molecule polypeptide compounds that can block PD-1/PD-L1 binding and have high anti-tumor activity. Methods Using the 5-peptide compound database constructed by our team and the three-dimensional structure data of the PD-L1 protein downloaded from the protein database (PDB), the peptide compounds obtained by flexible molecular docking using Molecular Operating Environment software. The molecular dynamics calculation and analysis of the compounds with the highest GWVI/WSA (generalized Born volume integral/weighted surface area) free energy change value (ΔG) were carried out, including the root mean square deviation (RMSD) of the position change of the weight atom and the interaction energy (the sum of Lennard-Jones potential and Coulombic energy). The blocking effect of ligand compounds on PD-1/PD-L1 binding was analyzed by the homogeneous time-resolved fluorescence (HTRF) technique. The co-culture system of Jurkat T lymphocytes and melanoma B16-F10 cells was established to explore the effect of compound ligands on the killing effect of T cells on tumors and the effect on the secretion level of IL-2 in the co-culture supernatant. Results The selected polypeptide compounds were analyzed by molecular dynamics, and the binding of RGGHA to RGGHH and PD-L1 was stable. There are many kinds of interactions between RGGHA and PD-L1，which can compete with PD-1 ligands to bind Asp122, Try123 and Lys124 sites of the PD-L1 protein and block PD-1/PD-L1 signal transduction. HTRF experiments showed that the binding inhibition rate of RGGHA to PD-1 and PD-L1 was 58.38%, and that of RGGHH was 42.73%. In addition, we established a co-culture system of Jurkat T lymphocytes and melanoma B16-F10 cells to explore the immunostimulatory effect and mechanism of peptides. The results show that RGGHA and RGGHH can significantly increase the secretion level of IL-2, improve the killing ability of T cells against tumor cells and activate the tumor immune microenvironment. Conclusions The study found that the polypeptide compounds RGGHA and RGGHH can effectively block the PD-1/PD-L1 interaction and reactivate the anti-cancer immune response, which can be used as lead compounds for new drug development.

Objective:To explore the clinical effects of scalp flaps pedicled with superficial temporal artery parietal branch in repairing facial destructive burn wounds.Methods:A retrospective observational study was conducted. From January 2016 to December 2021, 15 patients with facial destructive burns who met the inclusion criteria were admitted to Zhengzhou First People's Hospital, including 11 males and 4 females, aged 22 to 79 years. Two patients were complicated with unilateral eyeball destructive burns, two patients were complicated with unilateral auricle defects, eight patients were complicated with lip and cheek defects, and three patients were complicated with lip, cheek, and unilateral nasal alar defects. The burn wound areas ranged from 9 cm×6 cm to 13 cm×10 cm. The scalp flaps pedicled with superficial temporal artery parietal branch, with the area of 10 cm×7 cm to 15 cm×11 cm, were designed, excised, and transferred for repairing burn wounds. The secondary wounds at the donor sites were repaired with medium-thickness scalp grafts. According to patient's needs, the hairs grew at the facial transplanted flap were removed by laser at 2 weeks after the flap was completely viable, or the expanded scalp flap was used to treat the secondary alopecia in the flap donor area of the head at 3 months after the primary wound repair. The survival of the flap/skin graft and the wound healing of the donor and recipient areas after the primary wound repair were recorded. During the follow-up, the appearance of the flap, the scar hyperplasia at the suture site, the repair effect of facial functional parts, the treatment effects of laser hair removal and secondary alopecia treatment at the flap donor site were observed; the patient's satisfaction with the overall repair effect was inquired.Results:After the primary wound repair, all the flaps transplanted to the burn wounds and the skin grafts transplanted to the secondary wounds of the flap donor sites survived well, and the wounds at the donor and recipient sites of flap healed well. The color, texture, and thickness of flap were basically the same as those of normal facial skin, and the scar at the suture site was slight during 3 to 18 months of follow-up period after the primary wound repair. In 11 patients complicated with lip defects, the oral integrity, and the opening and closing functions of mouth were restored with the mouth opening being 2.0-2.5 cm and no microstomia; all the patients could carry out basic language communication, 8 of them could take regular food, and 3 of them could take soft food. The wounds in two patients with unilateral eyeball destructive burns were repaired. In 2 patients complicated with auricle defects, the wounds were repaired, and the external auditory canals were normal. In 3 patients complicated with unilateral nasal alar defects, their noses had poor appearance with reduced nostrils. No hair growth was observed in the facial flap sites after treatment of laser hair removal in 8 patients. Five patients were successfully treated with expanded scalp flaps for secondary alopecia in the flap donor area of the head. The patients were all satisfied with the overall repair effect.Conclusions:The scalp flap pedicled with superficial temporal artery parietal branch has abundant blood supply and is suitable for repairing the wounds in facial destructive burns. It is easy to transfer and can better restore the appearance and function of the recipient area with minimal damage to the flap donor area, which is worthy of clinical promotion.

The rapid screening of tumor markers is a challenging task for early diagnosis of cancer. This study aims to use highly sensitive chemiluminescent protein microarray technology to efficiently screen a variety of low abundance tumor related markers. A new material, termed integrated polydimethylsiloxane modified silica gel (iPDMS), was obtained by adding a surface polymerization initiator with olefin end to the conventional polydimethylsiloxane, and fixing into the three-dimensional structure of polydimethylsiloxane by thermal crosslinking through silicon hydrogen bonding. In order to make the iPDMS material resistant to non-specific protein adsorption, a poly(OEGMA) polymer brush was synthesized by surface-initiated atom transfer radical polymerization at the active initiation site. Finally, 20 tumor-related antigens were printed into the specific areas of the microarray by high-throughput spray printing technology, and assembled into 48-well detection microtiterplates of the iPDMS microarray. It was found the VEGFR and VEGF121 autoantibodies that obtained from 8 common tumors (breast cancer, lung cancer, colon cancer, gastric cancer, liver cancer, leukemia, lymphoma and ovarian cancer) can be used as potential tumor markers. The chemiluminescence labeled iPDMS protein microarray can be used for the screening of tumor autoantibodies at early stage.
Adsorption ; Autoantibodies ; Dimethylpolysiloxanes ; Protein Array Analysis ; Silica Gel ; Surface Properties

Plant trichomes are special structures that originate from epidermal outgrowths. Trichomes play an important role in plant defense against pests and diseases, and possess economic and medicinal values. Study on molecular mechanism of plant trichomes will contribute to the molecular design breeding and genetic improvement of crops. In recent years, the regulation mechanism of trichome development has been basically clarified in the model plant Arabidopsis thaliana, while great progresses are also found in other plant species. In this review, we focus on the developmental regulation of trichome formation from gene and phytohormones levels in Arabidopsis and cotton (with unicellular trichomes), as well as in tomato and Artemisia annua (with multicellular trichomes). The research progress associated with trichomes is also introduced in other typical monocotyledons and dicotyledons. Finally, the research and application of plant trichomes are prospected.
Arabidopsis/genetics* ; Gene Expression Regulation, Plant ; Gossypium/genetics* ; Lycopersicon esculentum ; Plant Growth Regulators/metabolism* ; Trichomes/genetics*

BACKGROUND: This study aimed to analyse the epidemiological characteristics of bacillary dysentery (BD) caused by Shigella in Chongqing, China, and to establish incidence prediction models based on the correlation between meteorological factors and BD, thus providing a scientific basis for the prevention and control of BD. METHODS: In this study, descriptive methods were employed to investigate the epidemiological distribution of BD. The Boruta algorithm was used to estimate the correlation between meteorological factors and BD incidence. The genetic algorithm (GA) combined with support vector regression (SVR) was used to establish the prediction models for BD incidence. RESULTS: In total, 68,855 cases of BD were included. The incidence declined from 36.312/100,000 to 23.613/100,000, with an obvious seasonal peak from May to October. Males were more predisposed to the infection than females (the ratio was 1.118:1). Children < 5 years old comprised the highest incidence (295.892/100,000) among all age categories, and pre-education children comprised the highest proportion (34,658 cases, 50.335%) among all occupational categories. Eight important meteorological factors, including the highest temperature, average temperature, average air pressure, precipitation and sunshine, were correlated with the monthly incidence of BD. The obtained mean absolute percent error (MAPE), mean squared error (MSE) and squared correlation coefficient (R) of GA_SVR_MONTH values were 0.087, 0.101 and 0.922, respectively. CONCLUSION From 2009 to 2016, BD incidence in Chongqing was still high, especially in the main urban areas and among the male and pre-education children populations. Eight meteorological factors, including temperature, air pressure, precipitation and sunshine, were the most important correlative feature sets of BD incidence. Moreover, BD incidence prediction models based on meteorological factors had better prediction accuracies. The findings in this study could provide a panorama of BD in Chongqing and offer a useful approach for predicting the incidence of infectious disease. Furthermore, this information could be used to improve current interventions and public health planning.

Objective To explore the effect of umbilical cord mesenchymal stem cells with positive human leukocyte antigen(HLA)-G on inducing the production of regulatory T cells(Treg) in vitro.Methods Umbilical cord mesenchymal stem cells were isolated from umbilical cord of neonates. PEGFP-N1-HLA-G plasmid was transfected into the human umbilical cord mesenchymal stem cells by liposome transfection, as PEGFP-N1-HLA-G group. PEGFP-N1 empty vector plasmid was transfected into the human umbilical cord mesenchymal stem cells, as PEGFP-N1 group. The human umbilical cord mesenchymal stem cells without empty vector under the same conditions were set as blank control group. Markers of the umbilical cord mesenchymal stem cells were detected using flow cytometry. The expression of HLA-G protein in each group of cells was identified by Western Blot. After mixed-culturing with CD4+T cells in peripheral blood of healthy subjects for 24 h and 48 h, the proportion of CD4+CD25+Foxp3+Treg in total T cells of each group was detected by flow cytometry. Results CD45, CD34 and HLA-DR presented negative expression on umbilical cord mesenchymal stem cells, while CD29, CD44 and CD105 presented positive expression. HLA-G protein could be expressed in the PEGFP-N1-HLA-G group, which had statistically significant difference compared with the blank control group and PEGFP-N1 group (both P<0.01). After PEGFP-N1-HLA-G group and CD4+T cells were mixed-cultured for 24 h and 48 h, CD4+CD25+Foxp3+Treg accounted for (15.3±1.9)% and (14.3±2.1)% of the total T cells respectively, both of which presented statistically significant difference compared with the blank control group and PEGFP-N1 group (all P<0.05). Conclusions Umbilical cord mesenchymal stem cells with HLA-G gene modified can effectively induce the production of CD4+CD25+Foxp3+Treg in vitro.

BACKGROUND:There is a very close relationship between osteoporosis and periodontal disease in postmenopausal women, but the mechanism remains unclear. OBJECTIVE:To investigate the expression of nuclear factor-κB in the alveolar bone andinterleukin-17 in the serum and gingiva in the mouse model of osteoporosis caused by ovariectomy. METHODS:Female mice aged 3 months were randomly divided into ovariectomy and sham operation groups. At 6 months after surgery, the mouse models were evaluated histologically on the submandibular bone and thigh bone stained with hematoxylin and eosin. In the submandibular bone, the expression levels of OCN and Runx2 were detected by RT-PCR, and the expression level of nuclear factor-κB was detected by immunohistochemical staining and western blot assay. Besides, the expression level of interleukin-17 in the serum and gingival homogenate was evaluated using Cytometric Beads Array. RESULTS AND CONCLUSION:The thigh bone in the ovariectomy group revealed the thin cortical bone, enlarged marrow cavity, and increased resorption lacunae, as well as fewer, thinner trabeculae with lower density and irregular structure. Compared with the sham operation group, the expression levels of OCN and Runx2 in the alveolar bone were decreased in the ovariectomy group. The activation of nuclear factor-κB (P65)appeared with P65 positive expression in the submandibular bone in the ovariectomy group, and the relative expression level was higher than that in the sham operation group. The serum level of interleukin-17 in the ovariectomy group was higher than that in the sham operation group, but the level in the gingival tissue showed no significant difference between the two groups. These results indicate that estrogen deficiency after ovariectomy can activate nuclear factor-κB signal pathway to play a role in periodontal osteolysis. However interleukin-17 in the local periodontal tissue may not be a key cytokine to damage the periodontal tissue.

Objective To determine the expression and clinical value of neuropilin-1 (NRP-1) in hepatocellular carcinoma (HCC).Methods One hundred and fifty-one cases of HCC tissues and 89 cases of healthy liver tissues were chosen to compare the expression of NRP-1 by immunohistochemistry.Then the relationships between different clinical factors and the expression of NRP-1 were analyzed by univariate and multivariate statistical analysis.Moreover,the survival rates were compared by survival analysis between different expressions of NRP-1 in HCC patients.Results Eleven cases were lost to follow-up or died for non HCC disease,and the effective cases in the final study were 140 cases.The positive expression rates of NRP-1 in HCC and normal liver tissues were 65.00％ and 35.96％ respectively,and the difference was statistically significant (x2 =18.843,P ＜0.001).According to the expression level of NRP-1,140 patients with HCC were divided into negative expression group (n =49) and positive expression group (n =91).Univariate analysis showed that the expression of NRP-1 in HCC was correlated with tumor number (x2 =8.025,P =0.005),TNM stage (x2 =26.467,P ＜ 0.001),differentiation degree (x2 =15.296,P ＜ 0.001),portal vein invasion (x2 =9.054,P =0.003) and hepatic vein invasion (x2 =5.928,P =0.015).Multivariate statistical analysis showed that TNM stage (OR =1.392,95％ CI:1.121-1.730,x2 =8.950,P =0.003),differentiation degree (OR =1.469,95％ CI:1.102-1.958,x2 =6.862,P =0.009),portal vein invasion (OR =1.829,95％ CI:1.157-2.893,x2 =6.665,P =0.010) and hepatic vein invasion (OR =2.161,95％ CI:1.172-3.987,x2 =6.084,P =0.014) were important factors for NRP-1 expression.The median survival time of NRP-1 negative HCC patients was significantly longer than that of positive group (44 months vs.23 months),and the difference was statistically significant (x2 =21.922,P ＜0.001).Conclusion NRP-1 is over-expression in HCC tissue and related to the malignant progress of HCC,and this suggests poor prognosis in patients with HCC.

Objective To observe the clinical effect ofTongfu-Xiefei decoction for the patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD) with phlegm-heat syndrome.Methods A total of 78 AECOPD patients were selected from June 2015 to December 2016 in the respiratory department of our hospital, and the patients were randomly divided into two groups according to the random number table method, 39 patients in each group. The control group was treated with convertional western medicine and placebo decoction. Treatment group was treated with convertional western medicine and true decoction. Both groups were treated for 2 weeks and fellowed up for 4 weeks. The symptoms score, SGRQ scores, blood gas analysis were observed and analyzed. The clinical curative effect and adverse evevts were observed.Results After treatment, the symptom cough (2.34 ± 1.11vs. 2.67±1.05,t=2.277), expectoration (1.91 ± 1.41vs. 2.35 ± 1.83, t=2.106), asthmoid (1.73 ± 1.15vs. 2.04 ± 1.53,t=2.214) scores in the treatment group were significantly lower than the control group (P<0.05). The SGRQ scores (42.47 ± 17.24vs. 59.95 ± 17.58,t=2.112) in the treatment group was significantly lower than the control group (P<0.05). The total effective rate in the treatment group was 91.9% (34/37) and 84.4% in the control group (27/32). The difference between the 2 groups was statistically significant (χ2=4.578,P=0.040).Conclusions TheTongfu-Xiefei decoction can effectively alleviate the symptoms and signs, improve the blood gas analysis index, and the long-term curative effect for AECOPD patients with phlegm-heat syndrome.

Objective To investigate the risk of hepatitis B virus (HBV) infection in the children born to HBV‐infected parent (s) after in vitro fertilization (IVF) .Methods From January 2008 to December 2013 , in the reproductive medicine center of the Third Affiliated Hospital of Zhengzhou University ,the medical history of 221 mothers by IVF were collected .Inclusion criteria were at least one of the parents was positive for hepatitis B surface antigen (HBsAg) ,and HBV DNA<1 × 104copies/mL . Hepatitis B vaccination status and HBV infection status of children were followed by telephone .Results One hundred and seventy‐two cases were followed up successfully ,and 49cases were lost .Two hundred and twenty‐four cases of children were HBsAg negative ,but one of them showed hepatitis B virus e antibody and hepatitis B virus core antibody positive .Conclusions The vertical transmission risk of HBV infection of children born to HBV‐positive patent(s) by IVF did not increase .