GRP78,as endoplasmic reticulum resident chaperone,is highly induced by a variety of tumor microenvironmental stresses,such as hypoxia and glucose deprivation.GRP78 is implicated in tumor cell proliferation,apoptosis,invasion,metastasis and angiogenesis.Recent evidence indicates that GRP78 levels is correlated with pathological grade,stage and prognosis for the majority of solid tumors.In addition,GRP78 plays an important role in drug tolerance and knockdown of GRP78 has been shown to sensitize malignant cells.GRP78 could not only be a good biomarker to predict cancer progression and chemo-responsiveness,but also an appealing target for the development of a more selective chemotherapy.

ner. Furthermore, IL-10 and TGF-β1 showed a synergic effect. Conclusion IL-10 and TGF-β1, the cytokines of regulatory T cells, suppress the formation and bone resorption function of the osteoclasts.

Objective:To construct a RNAi lentiviral vector targeting rat CD80 gene and detect its effect of gene silencing in NRK and IEC6 cells.Methods:The effective sequence of siRNA targeting rat CD80 gene was confirmed in our previous work.Oligo-DNA fragment containing short hairpin frame was synthesized and reannealed,and then cloned into pGCSIL-GFP lentiviral expression vector.PCR and sequencing analysis were made for verifying the positive clones.The virus packaging plasmids were transfected into 293T cells to harvest shRNA lentivirus.After infection in NRK and IEC6 cells,Real-time PCR was performed to determine the expressing level of CD80.Results:PCR and sequencing revealed that shRNA plasmids was correctly constructed.Virus with a titer of 4×10~8 TU/ml was successfully packaged.CD80 expression in NRK and IEC6 cells could be knockdown by virus infection as characterized by 66.9% and 63.5% decrease of CD80 mRNA in NRK and IEC6 cells respectively,compared with negative control lentivirus.Conclusion:The recombinant lentiviral shRNA expressing vector targeting rat CD80 gene has been successfully constructed and packaged.CD80 mRNA could be down-regulated availably in NRK and IEC6 cells.

Objective To observe the clinical effects of cognitive behavioral therapy (CBT) combined with pulmonary rehabilitation on the treatment of patients with pneumoconiosis. Methods A total of 108 patients with pneumoconiosis were selected as the research subject using convenient sampling method. They were randomly divided into control group and CBT group, with 54 cases in each group. Patients in the control group were treated with conventional symptomatic treatment and pulmonary rehabilitation treatment for 12 weeks. While patients in the CBT group were treated with CBT treatment in addition to treatments of the control group. The therapeutic effect was analyzed. Results Before treatment, there was no statistical difference on score of Montreal Cognitive Assessment (MoCA), percentage of forced expiratory volume in one second to the predicted value (FEV₁%), forced expiratory volume in one second to force vital capacity ratio (FEV₁/FVC%), score of the Modified Medical Research Council Dyspnea Scale (mMRC), six minute walk distance (6MWD), Hospital Anxiety and Depression Scale (HADS) subscale scores of anxiety (HADS-A) and depression (HADS-D), and scores of Chronic Obstructive Pulmonary Disease Assessment Test (CAT), St. George's Respiratory Questionnaire (SGRQ), and serum levels of leptin, interleukin-6 (IL-6) and C-reactive protein (CRP) between the two groups (all P>0.05). After treatment, the score of MoCA, FEV₁%, FEV₁/FVC% and 6MWD increased (all P<0.05), while the scores of mMRC, HDS-A, HDS-D, CAT and SGRQ decreased (all P<0.05), and the levels of serum leptin, IL-6 and CRP decreased (all P<0.05) in the CBT group compared with the control group. Conclusion The combined treatment of CBT and pulmonary rehabilitation can better improve the cognitive ability, respiratory function, motor function, negative emotion and quality of life, and alleviate inflammatory response in patients with pneumoconiosis. It is of certain clinical application value.

Objective:To explore the clinical phenotype and genetic etiology of a child with Developmental epileptic encephalopathy type 104 (DEE 104).Methods:A child who had presented at the Children′s Medical Center of the Affiliated Hospital of Guangdong Medical University in February 2021 for recurrent seizures over 1 month was selected as the study subject. Clinical data of the child was collected. Peripheral blood samples of the child and his parents were collected and subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing.Results:The child, a five-month-old male, had presented with frequent focal seizures with severe developmental retardation from infancy. Physical examination showed emaciation, microcephaly, oblique palpebral fissures, Stahl′s ears, and hypotonia in the limbs. Electroencephalogram revealed multi-focal sharp waves, slow waves and slow spinal waves. Cranial magnetic resonance imaging revealed enlargement of bilateral lateral ventricles and the third ventricle, along with widening of brain sulci, fissure and cisterna. WES revealed that he had harbored a heterozygous c. 2401C>T (p.His801Tyr) missense variant of the ATP6V0A1 gene. Sanger sequencing showed that both of his parents were of the wild type. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was predicted to be likely pathogenic (PS2+ PM2_Supporting+ PP3). The proband was diagnosed with DEE 104. Early treatment with sodium valproate has failed, but the child had become seizure free after the addition of levetiracetam and topiramate. He still had abnormal EEG discharges and severe psychomotor retardation. Combining our case and a review of literature, DEE104 is mainly caused by de novo heterozygous variants of the ATP6V0A1 gene with an autosomal dominant inheritance. The patients may show refractory epilepsy and severe global developmental delay from infancy. Conclusion:The c. 2401C>T (p.His801Tyr) variant probably underlay the DEE104 in this child.

Objective:To evaluate the oncologic outcomes of different laparoscopic radical hysterectomy.Methods:From January 2011 to December 2014, the laparoscopic operation cases of cervical cancer at stage Ⅰb1, Ⅰb2, Ⅱa1 and Ⅱa2, including the histologic subtypes of squamous-cell carcinoma, adenocarcinoma and adenosquamous carcinoma, were collected in five clinical centers. The data were divided into two groups according to the surgical procedures, that is, modified laparoscopic-vaginal radical hysterectomy (mLVRH) and total laparoscopic radical hysterectomy (TLRH). The overall survival rate (OS), disease-free survival rate (DFS) at 5 years were retrospectively analyzed in this study.Results:There were 674 cases in total, including 377 cases of mLVRH, 297 cases of TLRH. (1) The OS at 5 years: the mLVRH was 96.1% and the TLRH was 92.0%, and the mLVRH was higher than that of TLRH （ P=0.010）. Stratify analysis， including stage of disease （Ⅰb1 and Ⅱa1）， histologic subtypes （squamous-cell carcinoma, adenocarcinoma）， lymph node metastasis， revealed that, ① Stage of disease: in stage Ⅰb1, the OS at five years of mLVRH was higher than that in TLRH group （98.6% vs 93.6%, P=0.012）. In stage Ⅱa1, there was significant difference between the two groups, the OS at five years of mLVRH and TLRH were 93.6% and 77.6% （ P=0.007）. ② Histologic subtypes: for the OS at five years of squamous-cell carcinoma, mLVRH and TLRH were 96.1% and 92.3%, and there was significant difference （ P=0.046）； for adenocarcinoma, the OS at five years were 91.0% and 88.6%， and there was no difference between two groups （ P=0.230）. ③ Lymph node metastasis： the mLVRH and TLRH with lymph node metastasis, the OS at five years were 98.6% and 96.4%; the mLVRH and TLRH without lymph node metastasis, the OS at five years were 89.3% and 80.8%. There were no significant differences between the two groups,respectively ( P=0.156， P=0.093）. (2) The DFS at 5 years: there was no significant difference between mLVRH and TLRH (94.1% vs 90.9%, P=0.220). Stratify analysis for stage of disease, the mLVRH group was higher than that in the TLRH group in stage Ⅰb1 (97.0% vs 92.8%, P=0.039). However, for stage Ⅱa1, there was no significant difference between mLVRH and TLRH group （88.2% vs 75.8%, P=0.074）. Conclusions:The results of this retrospective study indicated that different laparoscopy surgical procedures had diverse oncologic outcomes. The OS at 5 years of the mLVRH is superior to the TLRH. The DFS at 5 years in Ⅰb1 stage, the mLVRH is higher than the TLRH. Therefore, the modified laparoscopy is still an alternative surgery for early cervical cancer patients when following the principle of no-tumor-exposure.