Objective:To investigate the role of hyaluronic acid-mediated motion receptor(HMMR)in the malignant progression of esophageal squamous cell carcinoma(ESCC)cells and its potential molecular mechanisms.Methods:8 samples of ESCC tissues and adjacent paracancerous tissues surgically removed at the Fourth Hospital of Hebei Medical University between January 2018 and December 2020,as well as ESCC cells KYSE-30 and KYSE-150,were collected.Western blotting(WB)and immunohistochemistry(IHC)were used to detect the expression of HMMR in ESCC tissues.RNA interference was used to knock down HMMR expression in KYSE-30 and KYSE-150 cells,and qPCR and WB were used to detect the knockdown effect.The effects of HMMR knockdown on the proliferation and invasion abilities of ESCC cells were detected by CCK-8 assay and Transwell assay,respectively.4-nitroquinoline 1-oxide(4NQO)was used to induce carcinogenesis in mice and establish an ESCC model.H-E staining was used to observe the morphological changes of esophagus,and IHC was used to analyze the expressions of HMMR,FAM83D(family with sequence similarity 83 member D),E-cadherin and N-cadherin in tissues of different degrees of carcinogenesis in mice.Results:The expression level of HMMR in human ESCC tissues was significantly higher than that in adjacent paracancerous tissues(all P<0.05).After HMMR knockdown,the proliferation and invasion abilities of KYSE-30 and KYSE-150 cells were significantly reduced(P<0.05 or P<0.01),and the expression level of FAM83D also decreased(all P<0.01).In nude mouse tumor experiment,the body weight of mice in the 4NQO group was lower than that of the control group(all P<0.05).The results of IHC staining showed that HMMR was highly expressed in tumor tissues(P<0.05),and the expression of HMMR in high-grade intraepithelial neoplasia(HGIN)tissues was significantly higher than that in low-grade intraepithelial neoplasia(LGIN)tissues(P<0.001).HMMR was positively correlated with the expressions of FAM83D and N-cadherin(r=0.724,0.870,all P<0.001),and negatively correlated with the expression of E-cadherin(r=-0.714,P<0.001).Conclusion:HMMR is highly expressed in ESCC tissues and may promote the progression of ESCC by up-regulating FAM83D expression.

This study was performed to investigate the variation characteristics and functions of Siglec-9 immune checkpoint in pulmonary tuberculosis.R language was used for bioinformatics analysis of GSE83456 chip data.Total of 48 patients with confirmed pulmonary tuberculosis(PTB)and 46 healthy controls were included.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the mRNA expression of Siglec-9 in peripheral blood;flow cytometry was used to detect the proportion of Siglec-9+CD4+T cells.The levels of TNF-α,IL-6,IFN-γ and IL-4 were detected by Cytometric Bead Array(CBA).Furthermore,the correlation of the proportion of Siglec-9+CD4+T cells with TNF-α and other cytokines were analyzed.Bioinformatics analysis showed that Siglec-9 was significantly increased in patients with PTB(P＜0.001),and was related to TNF-α/NF-κB,IL-6/JAK/STATA3,PI3K/AKT/mTOR signal pathways.Experimental data showed that the proportion of Siglec-9+CD4+T cells was significantly increased in patients with PTB(P＜0.001)and negatively correlated with the levels of TNF-α and other cytokines.In conclusion,the higher levels of Siglec-9 mRNA and Siglec-9+CD4+T cells in PTB may inhibit the function of CD4+T cells and participate in the occurrence and development of PTB.

This study was performed to investigate the variation characteristics and functions of Siglec-9 immune checkpoint in pulmonary tuberculosis.R language was used for bioinformatics analysis of GSE83456 chip data.Total of 48 patients with confirmed pulmonary tuberculosis(PTB)and 46 healthy controls were included.Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the mRNA expression of Siglec-9 in peripheral blood;flow cytometry was used to detect the proportion of Siglec-9+CD4+T cells.The levels of TNF-α,IL-6,IFN-γ and IL-4 were detected by Cytometric Bead Array(CBA).Furthermore,the correlation of the proportion of Siglec-9+CD4+T cells with TNF-α and other cytokines were analyzed.Bioinformatics analysis showed that Siglec-9 was significantly increased in patients with PTB(P＜0.001),and was related to TNF-α/NF-κB,IL-6/JAK/STATA3,PI3K/AKT/mTOR signal pathways.Experimental data showed that the proportion of Siglec-9+CD4+T cells was significantly increased in patients with PTB(P＜0.001)and negatively correlated with the levels of TNF-α and other cytokines.In conclusion,the higher levels of Siglec-9 mRNA and Siglec-9+CD4+T cells in PTB may inhibit the function of CD4+T cells and participate in the occurrence and development of PTB.

Objective:To observe the effect of Gongyanping Capsule on genital tract infected by ureaplasma urealyticum and fertility of mice, and to explore its mechanism.Methods:One hundred female ICR mice were divided into normal control group, model control group, azithromycin group, the low and high-dose group of Gongyanping Capsule according to random number table method. Except for the normal control group, the other groups were infected with ureaplasma urealyticum to establish the reproductive tract inflammation model. The azithromycin group was given 40 mg/kg of azithromycin, the low and high-dose groups were given 50 and 100 mg/kg of Gongyanping Capsule respectively, the normal control group and model control group were given equal volume of normal saline, once a day, for 4 consecutive weeks. The fertility status of each group was recorded. HE staining was used to observe the pathological changes of the vaginal tissues of the mice in each group, and the content of monocyte chemoattractant protein 1 (MCP-1), IL-4, IL-12, TNF-α myeloperoxidase (MPO) and GSH-Px of the mice in each group were determined; RT-PCR and Western blot were used to determine the vagina level of mRNA and protein expressions of TLR4 and NF-κB.Results:Compared with the model control group, the birth time and the number of dead mice in the azithromycin group and the low and high dose groups of Gongyanping Capsule decreased ( P<0.05), and the number of born mice increased ( P<0.05). The level of MCP-1, MPO, IL-4, IL-12, TNF-α decreased ( P<0.05), the level of GSH-Px increased ( P<0.05), the expression of TLR4 mRNA (1.25±0.33, 2.97±0.92, 2.32±0.72 vs. 3.69±1.32), NF-κB mRNA (1.48±0.42, 2.91±0.99, 2.13±0.70 vs. 3.83±1.41) decreased ( P<0.05), the expression of TLR4 (0.63±0.13, 1.32 ± 0.34, 1.04 ± 0.33 vs. 1.63 ± 0.41), NF-κB (0.63 ± 0.14, 1.36 ± 0.32, 1.03 ± 0.30 vs. 1.94 ± 0.58) decreased ( P<0.05), and had a certain dose-dependence. Conclusion:Gongyanping Capsule has obvious therapeutic effect on genital tract mice infected by ureaplasma urealyticum, and can significantly improve the fertility of mice; the mechanism may be related to that Gongyanping Capsule could inhibit the vaginal TLR4/NF-κB pathway in mice.

Objective:To investigate the effect of enhancing the strength of the hamstring on the stability of the knee joint.Methods:Thirty patients with anterior cruciate ligament (ACL) tears were randomly divided into a training group ( n=15) and a control group ( n=15). After the injury′s edema stage, all of the subjects received the standard 6-stage rehabilitation training for ACL injury, including isokinetic exercise, isometric tension and contraction exercise, single or bipedal jumping, proprioception exercises and cardiovascular exercise. On the basis of that standard training, additional hamstring strengthening training was given to the training group. It involved three sessions of weight-bearing flexion of the knee joint six to eight times, at least five times a week for three months. All of the subjects underwent the passive relaxation test (PRT), knee function scoring (Lysholm scores) and weight-bearing MRI before and within 1 month after the training. Anterior shift of the tibia (TAS) was measured using weight-bearing magnetic resonance imaging (MRI). Results:Before the training there were no significant differences between the groups in terms of average PRT or Lysholm scores. After the training, the average PRT score in neither group had improved significantly. The average Lysholm scores of the training and control groups were not significantly different either, though both groups′ averages had improved significantly compared with before the training. The average tibial shifts were also significantly smaller than before the training, with the training group′s average significantly smaller than that of the control group.Conclusion:Increasing hamstring muscle strength can reduce tibial anteversion in the weight-bearing upright position and improve the stability of the knee joint after ACL injury.

Objective To investigate the effect of Salvianolic acid on endoplasmic reticulum stress (ERS) pathway in brain hippocampus of PAP mice. Methods Twenty PAP dual transgenic male mice were selected, they were randomly divided into a PAP mice model group and a Salvianolic acid group, 10 mice in each group; another 10 SPF grade C57BL/6J male mice were selected as a normal control group. In the Salvianolic acid group, 0.9% normal saline solution of Salvianolic lyophilized injection (400 g/L) of dosage 21 mg·kg-1·d-1 was injected intravenously through a tail vein of mice; the PAP mice model and normal control groups were given the same amount of 0.9% normal saline, and the therapeutic course was consecutive 4 weeks in the three groups. At the end of the 4th week, the Morris water maze test was carried out to observe the changes of escape latency, the third quadrant residence time (RTQ), entry angle into water and cross-platform times of mice in each group; amyloid precursor protein (APP) positive cell expression in cerebral hippocampus of mice were detected by immunohistochemistry; Western Blot was used to detect the expression level of PER like endoplasmic reticulum kinase-eukaryon initiation factor 2α-C/EBP homogenous protein (PERK-eIF2α-CHOP) pathway related proteins in hippocampus of mice. Results The escape latency of the PAP mice model group on the 1st to 5th day were significantly longer than those of the normal control group, although a downward trend was observed on the 5th day, it was still significantly longer than that of the model group (seconds: 58.41±2.36 vs. 28.60±10.15); compared with the PAP mice model group, the escape latency of Salvianolic acid group was shorter at each time point, and reached the shortest level on the 5th day (seconds: 31.97±8.36 vs. 58.41±2.36). In the PAP mice model group, the RTQ and the number of crossing platforms were significantly lower than those in the normal control group [RTQ (seconds): 8.27±2.95 vs. 15.97±7.33, numbers of crossing platforms (frequency/90 s): 0.70±0.95 vs. 2.70±0.48]; the entry angle was obviously greater than that of the normal control group [(47.94±4.68)°vs. (32.66±2.55)°, P < 0.05]. Compared with PAP mice model group, in Salvianolic acid group, the RTQ and number of crossing platform were significantly higher [RTQ (seconds): 13.57±1.86 vs. 8.27±2.95, number of crossing platforms (frequency/90 s):1.60±0.97 vs. 0.70±0.47], the entry angle was markedly smaller [(35.46±6.79)°vs. (47.94±4.68)°,P < 0.05]. The positive expression rate of APP and the protein expressions of CHOP, p-eIF2α in PAP mice model group were significantly higher than those in the normal control group [the positive rate of APP: (60.44±6.19)% vs. (21.05±5.87)%, CHOP protein expression (gray value): 3.09±0.07 vs. 1.46±0.09, p-eIF2αprotein expression (gray value): 0.98±0.09 vs. 0.47±0.06, all P < 0.01], the expression of PERK and p-PERK were lower than those in normal control group [PERK (gray value): 0.42±0.06 vs. 0.82±0.11, p-PERK protein expression (gray value): 0.98±0.09 vs. 0.64±0.10, both P < 0.01]; the positive expression rate of APP and protein expressions of CHOP, p-eIF2α in Salvianolic acid group were significantly lower than those in PAP mice model group [positive expression rate of APP: (33.09±10.33)% vs. (60.44±6.19)%, CHOP protein expression (gray value): 1.57±0.12 vs. 3.09±0.07, p-eIF2α protein expression (gray value): 0.80±0.07 vs. 0.98±0.09, all P < 0.01], while PERK and p-PERK expression were significantly higher than those in the model group [PERK (gray value): 0.89±0.12 vs. 0.42±0.06, p-PERK (gray value): 0.78±0.08 vs. 0.98±0.09, both P < 0.01]. Conclusion Salvianolic acid might work through the PERK-eIF2α-CHOP pathway to reduce the retention of APP in the hippocampus tissue of PAP dual-transgenic mice, thereby the learning ability of the mice is improved, and the progression of brain injury delayed.

OBJECTIVETo explore the safety and efficacy of pegylated recombinant human granulocyte colony-stimulating factor (PEG-rhG-CSF) in preventing chemotherapy-induced neutropenia in patients with breast cancer and non-small cell lung cancer (NSCLC), and to provide the basis for clinical application.

METHODSAccording to the principle of open-label, randomized, parallel-group controlled clinical trial, all patients were randomized by 1∶1∶1 into three groups to receive PEG-rhG-CSF 100 μg/kg, PEG-rhG-CSF 6 mg, or rhG-CSF 5 μg/kg, respectively. The patients with breast cancer received two chemotherapy cycles, and the NSCLC patients received 1-2 cycles of chemotherapy according to their condition. All patients were treated with the combination chemotherapy of TAC (docetaxel+ epirubicin+ cyclophosphamide) or TA (docetaxel+ epirubicin), or the chemotherapy of docetaxel combined with carboplatin, with a 21 day cycle.

RESULTSThe duration of grade 3-4 neutropenia in the PEG-rhG-CSF 100 μg/kg and PEG-rhG-CSF 6 mg groups were similar with that in the rhG-CSF 5 μg/kg group (P>0.05 for all). The incidence rate of grade 3-4 neutropenia in the PEG-rhG-CSF 100 μg/kg group, PEG-rhG-CSF 6 mg group, and G-CSF 5 μg/kg group were 69.7%, 68.4%, and 69.5%, respectively, with a non-significant difference among the three groups (P=0.963). The incidence rate of febrile neutropenia in the PEG-rhG-CSF 100 μg/kg group, PEG-rhG-CSF 6 mg group and G-CSF 5 μg/kg group were 6.1%, 6.4%, and 5.5%, respectively, showing no significant difference among them (P=0.935). The incidence rate of adverse events in the PEG-rhG-CSF 100 μg/kg group, PEG-rhG-CSF 6 mg group and G-CSF 5 μg / kg group were 6.7%, 4.1%, and 5.5%, respectively, showing a non-significant difference among them (P=0.581).

CONCLUSIONSIn patients with breast cancer and non-small cell lung cancer (NSCLC) undergoing TAC/TA chemotherapy, a single 100 μg/kg injection or a single fixed 6 mg dose of PEG-rhG-CSF at 48 hours after chemotherapy show definite therapeutic effect with a low incidence of adverse events and mild adverse reactions. Compared with the continuous daily injection of rhG-CSF 5 μg/kg/d, a single 100 μg/kg injection or a single fixed 6 mg dose of PEG-rhG-CSF has similar effect and is more advantageous in preventing chemotherapy-induced neutropenia.

Antineoplastic Agents ; adverse effects ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; Breast Neoplasms ; drug therapy ; Carboplatin ; administration & dosage ; adverse effects ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; Cyclophosphamide ; administration & dosage ; adverse effects ; Epirubicin ; administration & dosage ; adverse effects ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Humans ; Incidence ; Induction Chemotherapy ; Lung Neoplasms ; drug therapy ; Neutropenia ; chemically induced ; epidemiology ; prevention & control ; Polyethylene Glycols ; Recombinant Proteins ; administration & dosage ; Taxoids ; administration & dosage ; adverse effects

Objective To investigate the relationship between the nerve growth factor ( NGF ) induced hippocampal neuroregeneration and homeobox gene Lhx 8.Methods Seventy-two SD rats were divided into control group , transected group, NGF group, transected combined with NGF group after right fimbria-fornix transection and NGF intracerebroventricular injection . Real-time PCR and Western blotting were applied to detect the gene and protein expression of Lhx8 in each group.The choline acetyltransferase ( ChAT)/Lhx8 double labeled cells in subgranular zone ( SGZ) of hippocampus in each group were detected by immunofluorescence .Results The expression of Lhx8 gene and protein in the transected , NGF group and especially in the transected combined with NGF group was obviously higher than in the control group .The number of ChAT/Lhx8 double labeled cells in the NGF group and the transected combined with NGF group was obviously more than in the control group and transected group . Conclusion The hippocampal neuroregeneration which induced by NGF intracerebroventricular injection was associated with the higher expression of Lhx8.

Objective The aim is to observe the role and mechanism of estradiol ( E2 ) on the proliferation of rat hippocampal neural stem cells ( NSCs ) .Methods Twenty hippocampi from embryonic 17-day ( E17 ) SD rats were dissociated and plated into culture flasks with NSCs specific medium containing different concentrations of estradiol .The proliferation and the vitality of NSCs were detected by immunofluorescence against BrdU and MTT assay .The expression of estrogen receptors ( ERαand ERβ) was measured by immunofluorescence staining combined with Nestin double labeling . Results BrdU and MTT assay results showed that the cell number increased when the concentration of estradiol increased from 10 -10 to 10 -8 mol/L.The number of cell proliferation and the viability of cells were best at the concentration of 10 -8 mol/L compared to the other groups .However, when the estradiol concentration was increased from 10-8 to 10 -6 mol/L, the cell proliferative capacity declined gradually .Double immunofluorescence labeling showed that the two types of estrogen receptors ( ERαand ERβ) were expressed in the cultured hippocampal NSCs .Conclusion Estradiol promotes the proliferation of hippocampal NSCs in a certain concentration range , and ERαand ERβmay be involved in the estradiol-induced proliferation .

Objective To investigate the effect of Jagged1 on hippocampal radial glial cells (RGCs) proliferation and neuronal differentiation in vitro.Methods Hippocampal RGCs were cultured in vitro, the agonist Jagged1 and(or) inhibitor DAPT of Notch signaling were added into the culture medium , and then the cells were divided into control group , Jagged1 group, Jagged1 combined with DAPT group and DAPT group .CCK-8 regent was used to detect cells ’ vitality;immunofluorescent was used to detect the number of BLBP /Ki67 double labeled cells and differentiated microtubule associated protein-2(MAP-2) positive cells.Results Cell vitality in Jagged1 group was obviously higher than that of the other groups .The number of BLBP/Ki67 double labeled cells and differentiated MAP-2 positive cells were more than other groups.Conclusion Jagged1 promotes the proliferation and neuronal differentiation of hippocampal RGCs in vitro.