Multidrug resistant genes are highly expressed in hepatocellular carcinoma that seriousty affects the effect of chemotherapy. Screening of resistant genes from HCC cells and studying its mechanism of drug resistance will be helpful to improve the effecacy of chemotherapy for hepatocellular carcinoma. Here we described an alternative method called cyclical packaging rescue (CPR). First we constructed a retrovirus cDNA library of hepatoma cells and used it to infect fibroblasts. Then we added drugs to screen survival cells. The survival cells, stably integrated helper-free retroviral libraries, were recovered rapidly after transfection with plasmids expressing retroviral gag-pol and env genes. Through this method, retroviral RNAs were directly repackaged into new infectious virions. Recovered retroviral supernatant was then used to reinfect fresh target cells. When performed in concert with selection using functional assays, cDNAs regulating functional responses could be identified by enrichment through multiple rounds of retroviral library recovery and retransmission. Using CPR, we obtained several cDNAs. After a preliminary detection, we found Ribosomal protein S11 (RPS11), Ribosomal protein L6 (RPL6), Ribosomal protein L11 (RPL11), Ribosomal protein L24 (RPL24) possibly had drug resistant function.
Carcinoma, Hepatocellular ; genetics ; pathology ; Cell Line, Tumor ; DNA, Complementary ; Drug Resistance, Neoplasm ; genetics ; Gene Library ; Genetic Vectors ; Humans ; Liver Neoplasms ; genetics ; pathology ; Plasmids ; Retroviridae ; Ribosomal Proteins ; genetics ; metabolism ; Transfection

Objective To elucidate the expression of gankyrin in human gastric cancer cells and it's role in nimesulide induced apoptosis. Methods Four human gastric cancer cell lines including MKN28 (well differentiated), AGS (poorly differentiated), MKN45 (poorly differentiated), and SGC7901(moderately differentiated) were cultured and treated with nimesulide. Nimesulide induced growth inhibition and apoptosis of the cells were detected by methyl thiazolyl tetrazolium assay, and confirmed by flow cytometry. The expressions of gankyrin gene and protein were further assessed by real-time PCR and Western blotting. Results Gankyrin mRNA and protein were detected in all four human gastric cancer cell lines. The proliferations of AGS and SGC7901 cell lines were significantly suppressed by nimesulide in a time-dose dependent manner. When treated with 400 μmol/L of nimesulide for 48 hours, the significant apoptosis was found in AGS cells (23.30%±2.50%) and SGC7901 cells (16.80%±1.55% ) in comparison with controls (0.57%±0.19% and 0.88%± 0.17%, respectively, all P values <0.01). Apoptosis of AGS cells induced by nimesulide was accompanied by a considerably decreased gankyrin expression that was more significant at 24 hours (0.0035±0.0014) and 36 hours (0.0980±0.0160) in comparison with controls (0.4690±0.1190, all P values<0.01). Conclusion Gankyrin expresses in human gastric cancer cell lines and may be involved in nimesulide induced apoptosis of AGS cells.

Objective:To investigate the effect of health belief model-based intervention on post-traumatic stress disorder and post-traumatic growth of parents of children with malignant lymphoma.Methods:A total of 90 malignant lymphoma children parents were randomly divided into study group and control group, with 45 cases in each group. The control group received routine nursing method, the study group carried out health belief model-based intervention. The Posttraumatic stress Checklist-Civilian version (PCL-C) and Post-traumatic Growth Inventory (PTGI) were used to evaluate the effect of intervention, respectively.Results:After intervention, the scores of PLC-C were significantly decreased and PTGI were significantly increased both in the intervention group and control group. After intervention, the scores of arousal symptoms avoidance/numbing, re-experiencing symptoms and total PCL-C scores were (7.26±1.13), (8.80±1.95), (5.67±1.16), (21.73±2.57) points in the study group, which were significantly decreased compared to (8.09±1.63), (9.92±3.05), (6.10±0.68), (24.10±3.78) points in the control group ( t values were 2.070-3.484, all P<0.05 or 0.01). After intervention, the scores of relating to others, personal strength and total PTGI scores were (21.66±4.66), (18.28±5.04), (73.93±8.20) points in the study group, which were significantly increased compared to (18.39±4.39), (16.43±2.71), (66.46±6.20) points in the control group ( t values were 3.415, 2.161, 4.877, all P<0.05 or 0.01). Conclusion:Health belief model-based intervention can effectively alleviate post-traumatic stress disorder and promote posttraumatic growth in parents with malignant lymphoma children.

Brucellosis is a zoonotic infectious disease caused by Brucella infection. So far, animal to animal Brucellosis has not been eradicated, and there is a lack of safe and effective human vaccine. Therefore, "early, combined, sufficient, and full course" drug treatment remains an important strategy in the management of human Brucellosis. The goal of treating brucellosis is to alleviate and shorten the symptom period, reduce complications, relapses, and chronicity. At present, although antibiotic treatment is effective for most patients, there are still some patients who experience treatment failure or later recurrence, so the treatment strategy for brucellosis urgently needs to be optimized. This article elaborates on the treatment principles, clinical treatment status, and future development trends of brucellosis, in order to provide references for optimizing drug treatment methods for brucellosis.

Objective:To investigate the impact of the birth of preterm infants on their family functioning, and to provide a reference for the formulation of targeted intervention programs.Methods:Based on the socio ecosystem theory, phenomenological research methods and objective sampling were used to conduct semi-structured interviews with 16 preterm infants in the neonatal intensive care unit (NICU) of Children′s Hospital of Soochow University from September 2023 to January 2024, who meet the inclusion and exclusion criteria, and the Colaizzi 7-step analysis method was used to analyze the interview data.Results:All 16 caregivers of the NICU preterm infants who participated in the interviews were the infants′ mothers, aged from 26 to 34 years. A total of 3 themes and 10 sub-themes were summarized. The micro system: physical and mental exhaustion coexisted with role growth of caregivers (physical and mental health was affected, positive experience after psychological adjustment, and adaptation to role change). The intermediate system: family relationship mode change (family satisfaction decreased, family difficulties affected family harmony, family communication content changed, family harmony and conflict coexisted, and family conflict resolution was actively resolved). The outer system: social support system needed to be improved (benefited from social support and not yet perceived external support).Conclusions:Medical staff can improve the multiple support system and provide continuous and personalized family support, so as to improve the family adjustment ability and improve the family functioning of preterm infants in the NICU.

Objective To compare the efficacy and safety of leflunomide (LEF) combined with medium/low dose corticosteroids and full dose of corticosteroids in the treatment of IgA nephropathy. Method Primary IgAN patients diagnosed by renal biopsy with 18?65 years old and eGFR≥30 ml·min?1·(1.73 m2)?1 and proteinuria>0.5 g/24 h were enrolled in a prospective controlled clinical study. They were randomly divided into leflunomide combined with medium/low dose corticosteroids (LEF group) and corticosteroids alone (steroid group). The primary outcomes were (1) end stage renal disease or dialysis (2) 50% increase in serum creatinine above the baseline. Secondary outcome was the remission of proteinuria. Results Ninety patients completed the follow?up. The 24?hour proteinuria at baseline were 2.00(1.10, 2.88) g and 1.87(1.13 ,3.08) g in LEF group and steroid group respectively. Compared with baseline, it was significantly decreased in both groups at 6 months [0.30(0.11, 0.93) g, 0.30(0.14, 1.33) g] and 12 months [0.30(0.09, 0.82) g, 0.32(0.14, 0.66) g], (P<0.05). Estimated glomerular filtration rate (eGFR) at baseline, 6 months and 12 months were (80.39 ± 28.56), (87.12±28.70) and (88.20±30.26) ml·min-1·(1.73 m2)-1. It was decreased in steroid group (P<0.05), while no significant difference was detected in LEF group[baseline (87.63 ± 27.35), 6 months (86.91 ± 32.45), 12 months (90.06 ± 30.00) ml·min-1·(1.73 m2)-1, P>0.05]. At 6 and 12 months, there was no significant difference in terms of 24?hour proteinuria, serum creatinine and eGFR (CKD?EPI) between groups (P>0.05). There was no statistically significant difference in adverse events between groups during the treatment (9/40 cases in LEF group and 11/50 cases in steroid group, P>0.05). The average follow?up was 79 months, and there was no difference in the renal prognosis between the two groups. Multivariate Cox regression analysis revealed that serum creatinine at baseline and renal interstitial inflammatory cell infiltration predicted the risk of the progress of IgA nephropathy. Conclusion Leflunomide plus medium/low dose corticosteroids has a similar effect as full dose of corticosteroids in IgA nephropathy and does not increase the risk for adverse events during the treatment.

OBJECTIVE: To diagnose the mycotic otitis media correctly and to explore the most adequate treatment for the disease. METHOD: Thirty-six inpatients (39 ears) with mycotic otitis media in Nanjing Drum Tower Hospital from Jan. 2003 to Dec. 2007 were analyzed retrospectively. Morphous of the fungi, the methods and efficacies of the treatment were analyzed respectively. RESULT: According to the fungal cultures, 27 ears were induced by mold fungus and 12 ears were induced by budding fungus. Among these 36 patients (39 ears), myringoplasty accompanied local antifungal cream were applied in one ear, mastoidectomy with canal wall down and/or tympanoplasty accompanied with oral antifungal medication were administrated in 35 ears, only oral antifungal drugs were used in 3 ears (the control ears of the bilateral mycotic otitis media, which was not treated by surgery). All of the patients were followed up for 3 to 36 months, otorrhea occurred in the patients who refused to oral antifungal medication for 3 weeks after the myringoplasty, then dry again by local antifungal cream, but otorrhea recurred 3 times within 2 years. Thirty-five patients (38 ears) acquired dry ear after surgery and/or oral antifungal drugs, but 2 of the 38 ears recurred separately at the fourth and sixth month after their surgeries, then dry again by irrigation with hydrogen peroxide and by administrating local antifungal cream for 3 weeks. CONCLUSION Otologists should elevate suspicion of mycotic otitis media when they meet patients with continuous otorrhea and patients who did not respond to the antibacterial treatment. The diagnosis based on microbiological findings, such as direct microscopy or fungal cultures should be done as soon as possible. If the otomycosis is decided, we suggest that topical treatment should be selected firstly, although most patients in present study were treated by surgery accompanied with oral antifungal medications. If there is obvious bone erosion, surgery is necessary to excise the pathological tissues, minificate the mastoid cavity and close the middle cavity in order to improve the hearing and prevent the infection from the outer ear.
Adult ; Aged ; Chronic Disease ; Female ; Humans ; Male ; Middle Aged ; Mycoses ; diagnosis ; therapy ; Otitis Media ; diagnosis ; microbiology ; therapy ; Retrospective Studies ; Treatment Outcome ; Young Adult

Objective:To explore the effect of extracellular signal-regulated kinase (ERK) inhibitor PD98059 on calpain-related proteins in the brain, and to understand the pathophysiological changes of calpain in cerebral ischemia/reperfusion injury (CIRI).Methods:Forty-two rats were divided into sham operation (Sham) group ( n = 6), model group ( n = 12), dimethyl sulfoxide (DMSO) control group ( n = 12), and PD98059 group ( n = 12) by random number table. The rat model of CIRI induced by cardiac arrest-cardiopulmonary resuscitation (CA-CPR) was reproduced by transesophageal electrical stimulation to induce ventricular fibrillation. In the Sham group, only the basic operations such as anesthesia, tracheal intubation, and arteriovenous catheterization were performed without CA-CPR. The rats in the DMSO control group and PD98059 group were injected with DMSO or PD98059 0.30 mg/kg via femoral vein, respectively, 30 minutes after the restoration of spontaneous circulation (ROSC), and rats in the Sham group and model group were given the same amount of normal saline. The duration of CPR, 24-hour survival rate and neurological deficit score (NDS) after ROSC were recorded. Hematoxylin-eosin (HE) staining and Nissl staining were used to observe the pathological changes of the cerebral cortex. The expressions of phosphorylated ERK (p-ERK), ERK, calpastatin, calpain-1, and calpain-2 were detected by Western blotting. The co-expression of p-ERK and calpain-2 was detected by double immunofluorescence. Results:There were no significant differences in the duration of CPR and 24-hour survival rate among all groups. In the model group, the nuclei of the cerebral cortex were obviously deformed and pyknotic, cells vacuoles and tissues were arranged disorderly, Nissl corpuscles were significantly reduced, NDS scores were also significantly reduced, level of ERK phosphorylation was increased, and calpain-2 protein was significantly up-regulated compared with the Sham group. There was no significant difference in the above parameters between the DMSO control group and the model group. After intervention with PD98059, the pathological injury of brain tissue was significantly improved, Nissl corpuscles were significantly increased, the NDS score was significantly higher than that in the model group [75.0 (72.0, 78.0) vs. 70.0 (65.0, 72.0), P < 0.05], the level of ERK phosphorylation and calpain-2 protein expression were significantly lower than those in the model group [p-ERK (p-ERK/ERK): 0.65±0.12 vs. 0.92±0.05, calpain-2 protein (calpain-2/GAPDH): 0.73±0.10 vs. 1.07±0.14, both P < 0.05], while there was no significant difference in the expressions of calpastatin and calpain-1 in the cerebral cortex among all the groups. Double immunofluorescence staining showed that p-ERK and calpain-2 were co-expressed in cytosol and nucleus, and the co-expression rate of p-ERK and calpain-2 in the model group was significantly higher than that in the Sham group [(38.6±4.3)% vs. (9.2±3.5)%, P < 0.05], while it was significantly lowered in the PD98059 group compared with the model group [(18.2±7.0)% vs. (38.6±4.3)%, P < 0.05]. Conclusions:ERK together with calpain-2 participated in CIRI induced by CA-CPR. PD98059 inhibited the expression of calpain-2 and ERK phosphorylation. Therefore, ERK/calpain-2 may be a novel therapeutic target for CIRI.

Objective: To investigate the clinical and immunological laboratory features, mutations in SH2D1A gene and SAP protein expression in four children of two families with X-linked lymphoproliferative disease type 1(XLP-1). Method: Four patients (Family A including Patient 1 and Patient 2, Family B including Patient 3 and Patient 4) and their maternal relatives were enrolled in this study. The clinical manifestation, EBV infection status and chest CT scan were analyzed. The absolute and relative numbers of lymphocyte subsets, T lymphocyte proliferative response, SAP protein expression were assessed by flow cytometry. Quantification of signal joint TCR rearrangementexcision circle (sjTRECs), CDR3 spectratyping of TCRvβ and gene mutation of SH2D1A were detected by PCR based on genomic DNA or cDNA. Result: Four male patients from two families were diagnosed with XLP-1. The ages of disease onset were more than 1 year, more than 1 year, more than 1 month and 6 months. The ages at diagnosis were nine years and ten months, sixteen years and eight months, fourteen years and ten months, four years and nine months. All patients had recurrent infections and EBV infection. Patients 1, 2, and 3 had agammaglobulinemia and Patient 4 had hypogammaglobulinemia. Chest CT scan showed all patients had atelectasis and pneumonia, and Patient 3 had bronchiectasis. Patient 3 was diagnosised as Burkitt lymphoma. For immunological function, all patients exhibited reduced CD4/CD8 ratios, increased numbers of exhausted T lymphocyte, decreased number of NK cell. The numbers of total B lymphocyte and naïve B lymphocyte were normal, but the number of memory B lymphocyte declined in all cases. Four patients′ copy numbers of sjTRECs were low and CDR3 spectratypings of TCRvβ showed mildly skewed. But their T lymphocyte proliferative response was normal. SAP protein expression in four cases were measured by flow cytometry. Two patients from Family A were absent and two patients from Family B showed decreased values. SH2D1A gene sequence analysis showed that the patients of Family A harbored a nonsense mutation (c.163 C>T; p.R55X) in exon 2. Their mother and two sisters were carriers. A missense mutation of SH2D1A gene (c.278 G>A; p.G93D) in exon 3 was found in the patients of Family B. The mother was carrier. Four patients remain survived, Patient 3 gave up treatment, other three patients received IVIG therapy. Conclusion Four patients with XLP-1 from two families characterized by agammaglobulinemia have an extreme vulnerability to Epstein-Barr virus (EBV) infection. The functions of T cell, B cell and NK cell are impaired at different stages. The detection of SAP protein and SH2D1A gene are the key methods for diagnosis of XLP-1.

The biological samples of oral genetic diseases and rare diseases are extremely precious. Collecting and preserving these biological samples are helpful to elucidate the mechanisms and improve the level of diagnose and treatment of oral genetic diseases and rare diseases. The standardized construction of biobanks for oral genetic diseases and rare diseases is important for achieving these goals. At present, there is very little information on the construction of these biobanks, and the standards or suggestions for the classification and coding of biological samples from oral and maxillofacial sources, and this is not conducive to the standardization and information construction of biobanks for special oral diseases. This consensus summarizes the background, necessity, principles, and key points of constructing the biobank for oral genetic diseases and rare diseases. On the base of the group standard "Classification and Coding for Human Biomaterial" (GB/T 39768-2021) issued by the National Technical Committee for Standardization of Biological Samples, we suggest 76 new coding numbers for different of biological samples from oral and maxillofacial sources. We hope the consensus may promote the standardization, and smartization on the biobank construction as well as the overall research level of oral genetic diseases and rare diseases in China.