Evaluation of the Effectiveness of In-Hospital Exercise Rehabilitation in Middle-Aged and Elderly Patients with Type 2 Diabetes Mellitus Combined with Sarcopenia Effect of Lifestyle Management on Disease Status in Patients with Type 2 Diabetes Mellitus.

OBJECTIVE: The aim of this study was to explore the effects of in-hospital exercise rehabilitation on glucose and lipid metabolism and healthy physical fitness in middle-aged and elderly patients with type 2 diabetes mellitus (T2DM) combined with sarcopenia, and to provide a reference for the effective implementation of exercise rehabilitation for middle-aged and elderly patients with T2DM combined with sarcopenia in healthcare institutions. METHODS: This study retrospectively included 122 patients with T2DM combined with sarcopenia treated at the General Hospital of Ningxia Medical University from August 2017 to August 2020 and randomly divided into a control group and an experimental group. The control group was given conventional treatment and the experimental group was given exercise rehabilitation in the hospital for 12 weeks to compare the indexes related to glucose and lipid metabolism and healthy fitness in the two groups. RESULTS: After the intervention, the experimental group showed significant decreases in fasting blood glucose (FBG), glycated hemoglobin (HbA1c), insulin resistance index (HOMA-IR), triglycerides (TG), total cholesterol (TC), low-density cholesterol (LDL-C) and body fat percentage (p < 0.05), while high-density cholesterol (HDL-C), grip strength, lower limb extension, lower limb flexion, peak oxygen uptake were significantly higher (p < 0.05) and were more significant at 12 weeks compared to the 6-week intervention (p < 0.05). However, there were no significant changes in any of the glucose metabolism indicators in the control group before and after the intervention. A two-way repeated measures ANOVA showed that at control baseline levels, HbA1c decreased significantly in the experimental group after both 6 and 12 weeks of intervention compared to the control group (p < 0.05). After 6 weeks of intervention, the experimental group showed a significant decrease in body fat percentage and a significant increase in grip strength. After 12 weeks of intervention, the experimental group showed an increase in glycemic control from 33.3% to 73.3%, a significant decrease in body fat percentage and a significant increase in grip strength, lower limb extension and lower limb flexion strength and peak oxygen uptake. CONCLUSION: In-hospital exercise rehabilitation can effectively improve the glycemic and lipid profiles of patients with T2DM combined with sarcopenia and enhance their health fitness, with good clinical rehabilitation effects.

Mechanism of Notch Signal Regulating Alveolar Epithelial Cell Autophagy after Infection with Klebsiella Pneumoniae.

BACKGROUND: The Notch signaling pathway regulates various cellular processes, including cell growth, inflammation response, and autophagy, thereby participating in the occurrence and development of various diseases. The present study aimed to investigate the molecular mechanism of Notch signaling in regulating alveolar type II epithelial cell viability and autophagy after Klebsiella pneumonia (KPN) infection. METHODS: KPN-infected human alveolar type II epithelial cells A549 (ACEII) were constructed. The autophagy inhibitor 3-methyladenine (3-MA) and Notch1 signaling inhibitor (DAPT) were used to pretreat A549 cells for 24 hours, 48 hours, and 72 hours before KPN infection. Real-time fluorescent quantitative PCR (qRT-PCR) and western blot assays were applied to detect the mRNA and protein expressions of LC3 and Notch1, respectively. ELISA was used to detect the levels of INF-γ, TNF-α, and IL-1ß in the cell supernatants. RESULTS: The results showed that KPN-infected A549 cells presented significantly upregulated Notch1 and autophagy-related protein LC3 levels, along with increased IL-1ß, TNF-α, INF-γ levels in a time-dependent manner. Autophagy inhibitor 3-methyladenine (3-MA) counteracted the promotive effects of LC3 and inflammatory cytokine levels in KPN-infected A549 cells; however, 3-MA did not influence Notch1 level. Notch1 inhibitor DAPT could suppress Notch1 and LC3 levels, thereby inhibiting inflammation response in KPN-treated A549 cells in a time-dependent way. CONCLUSIONS: KPN infection can activate the Notch signaling pathway and induce autophagy in type Ⅱ alveolar epithelial cells. Inhibiting the Notch signaling pathway may restrain KPN-induced A549 cell autophagy and inflammation response, shedding new insights for the treatment of pneumonia.

Preparation, characterization, and in-vitro cytotoxicity of nanoliposomes loaded with anti-tubercular drugs and TGF-ß1 siRNA for improving spinal tuberculosis therapy.

BACKGROUND: Tuberculosis (TB) represents a bacterial infection affecting many individuals each year and potentially leading to death. Overexpression of transforming growth factor (TGF)-ß1 has a primary immunomodulatory function in human tuberculosis. This work aimed to develop nanoliposomes to facilitate the delivery of anti-tubercular products to THP-1-derived human macrophages as Mycobacterium host cells and to evaluate drug efficiencies as well as the effects of a TGF-ß1-specific short interfering RNA (siRNA) delivery system employing nanoliposomes. METHODS: In the current study, siTGF-ß1 nanoliposomes loaded with the anti-TB drugs HRZ (isoniazid, rifampicin, and pyrazinamide) were prepared and characterized in vitro, determining the size, zeta potential, morphology, drug encapsulation efficiency (EE), cytotoxicity, and gene silencing efficiency of TGF-ß1 siRNA. RESULTS: HRZ/siTGF-ß1 nanoliposomes appeared as smooth spheres showing the size and positive zeta potential of 168.135 ± 0.5444 nm and + 4.03 ± 1.32 mV, respectively. Drug EEs were 90%, 88%, and 37% for INH, RIF, and PZA, respectively. Meanwhile, the nanoliposomes were weakly cytotoxic towards human macrophages as assessed by the MTT assay. Nanoliposomal siTGF-ß1 could significantly downregulate TGF-ß1 in THP-1-derived human macrophages in vitro. CONCLUSION: These findings suggested that HRZ-loaded nanoliposomes with siTGF-ß1 have the potential for improving spinal tuberculosis chemotherapy via nano-encapsulation of anti-TB drugs.

[Effect of germ cell Wdr1 deletion on the ovarian function of mice].

OBJECTIVE: To study the effect of Wdr1 deletion in germ cells on ovarian function of mice. METHODS: Oocyte-specific gene knockout mouse model was constructed by crossing Wdr1fl/flfemale mice with Cre recombinase transgenic male mice which was driven by a germ cell-specific promoter. Wdr1fl/-; Ddx4-Cre mice and control mice were sacrificed at 14 days, 28 days and 4 months after birth, whose ovaries were subjected to photography, paraffin sectioning and Hematoxylin-Eosin (HE) staining. The ovarian volume and follicular numbers were recorded at various time points. RESULTS: The ovarian volume of Wdr1 fl/-; Ddx4-Cre mice was slightly lower than that of the controls at 14 days. HE staining showed that primordial follicles, primary follicles and secondary follicles were slightly reduced compared with the control mice at 14 days. The ovarian volume of Wdr1 fl/-; Ddx4-Cre mice was significantly lower than that of the control mice at 28 days and 4 months. HE staining showed that all developmental follicles were significantly reduced compared with the control mice. CONCLUSION: Wdr1 gene deletion in germ cells can influence early ovarian function of mice and lead to premature ovarian failure.

Case report: A case of high virulence and multidrug resistant Klebsiella pneumoniae liver abscess in Ningxia, China.

RATIONALE: Highly virulent multidrug-resistant Klebsiella pneumoniae (KP) is becoming more and more common in clinical practice, especially the rise of carbapenem-resistant KP in clinical practice, resulting in the emergence of KP liver abscess in Ningxia, China. For the prognosis of liver abscess patients, it is particularly important to identify the types of pathogens and identify antibiotics that are sensitive to the pathogens. PATIENT CONCERNS: A 73-year-old man from China presents to our hospital with abdominal pain, jaundice and fever. Patients have no obvious cause of abdominal pain, abdominal distension, and abdominal pain is persistent. Abdominal examination showed hepatomegaly, no tenderness 2 cm from the right costal margin, abdominal distension and other general examinations did not have obvious abnormalities. He had no history of hypertension and diabetes, ERCP was performed for cholangiocarcinoma 1 year before the current visit, and no significant complications occurred. DIAGNOSES: His initial diagnosis was obstructive cholangitis, and computed tomographic images and liver drainage fluid bacterial culture and genetic polymerase chain reaction tests later determined that the patient had KP liver abscess. INTERVENTIONS: Drainage by liver catheter and antibiotic treatment for 7 weeks. OUTCOMES: The patient liver abscess is basically gone. LESSION: It is particularly important to optimize the diagnosis of liver abscess pathogens for timely and effective treatment of patients.

[Study on immune response in ICR mice by immunization with recombinant myophilin vaccine against Echinococcus granulosus].

OBJECTIVE: To investigate the immune response and the protection in mice induced by the recombinant myophilin protein of Echinococcus granulosus. METHODS: Thirty-six male ICR mice of 6-8 weeks old were randomly divided into groups A, B and C each with 12. The mice in the 3 groups were subcutaneously immunized with Eg myophilin protein, blank plasmid protein or PBS, respectively, by 3 times and challenged with protoscoleces of E. granulosus 2wk after the last vaccination. Mice were sacrificed 20wk after the infection, the hydatid cysts were collected for measuring the weight reduction. Spleens were obtained and the splenocytes were separated and cultured in vitro with EgAg or ConA stimulus for 4-5 h. The subsets of CD4+ and CD8+ T cells were measured by FACsort. The proliferation of splenocytes was determined by MTT method with blank plasmid and PBS as control. RESULTS: The average weight of the hydatid cysts in the immunized group decreased by 69.1% in comparison to the blank plasmid and PBS groups. The CD4+ subset [(29.7 +/- 0.9)%] and CD84+ subset [(9.7 +/- 0.8)%] in group A increased significantly than group C, [(11.6 +/- 1.4)%] and [(7.8 +/- 0.2)%] respectively (P < 0.01 or < 0.05). The ratio of CD4+/CD8+ subsets in group A (3.061 +/- 0.015) was also higher than group C (1.487 +/- 0.106) (P < 0.01). Without stimulation, the proliferation of T lymphocytes in group A(0.237 +/- 0.009) was higher than group C (0.159 +/- 0.005) (P < 0.01), with EgAg or ConA stimulus, it was also higher in group A than that of group C (P < 0.01). CONCLUSION: The recombinant myophilin protein of E. granulosus can induce the proliferation of splenocytes and Th1 response in mice, and the CD4+ T cells subset may bear a part in the induced protection against the challenge of protoscoleces.

Nomogram for the Prediction of Intrahospital Mortality Risk of Patients with ST-Segment Elevation Myocardial Infarction Complicated with Hyperuricemia: A Multicenter Retrospective Study.

PURPOSE: This study aimed to establish an accurate and easy predictive model for ST-segment elevation myocardial infarction (STEMI) patients with hyperuricemia, using readily available features to estimate intrahospital mortality risk. PATIENTS AND METHODS: This was a multicenter retrospective study involving the development of risk prediction models for intrahospital mortality among all STEMI patients with hyperuricemia from Zunyi Medical University Chest Pain Center's specialized alliance between January 1, 2016 and June 30, 2020. The primary outcome was intrahospital mortality. A total of 48 candidate variables were considered from demographic and clinical data. The least absolute shrinkage and selection operator (LASSO) was used to develop a nomogram. Concordance index values, decision curve analysis, the area under the curve (AUC), and clinical impact curves were examined. In this study, 489 patients with STEMI were included in the training dataset and an additional 209 patients from the 44 chest pain centers were included in the test cohort. B-type natriuretic peptides, α-hydroxybutyrate dehydrogenase (α-HBDH), cystatin C, out-of-hospital cardiac arrest (OHCA), shock index, and neutrophil-to-lymphocyte ratio were associated with intrahospital mortality and included in the nomogram. RESULTS: The model showed good discrimination power, and the AUC generated to predict survival in the training set was 0.875 (95% confidence interval, 0.825-0.925). In the validation set, the AUC of survival predictions was 0.87 (95% confidence interval, 0.792-0.947). Calibration plots and decision curve analysis showed good model performance in both datasets. A web-based calculator (https://bzxzmu.shinyapps.io/STEMI-with-Hyperuricemia-intrahospital-mortality/) was established based on the nomogram model, which was used to measure the levels of OHCA, neutrophil-to-lymphocyte ratio, shock index, α-HBDH, cystatin C, and B-type natriuretic peptides. CONCLUSION: For practical applications, this model may prove clinically useful for personalized therapy management in patients with STEMI with hyperuricemia.

Klebsiella pneumoniae infection inhibits autophagy by alveolar type II epithelial cells.

To investigate the molecular mechanism underlying the interaction between autophagosomes of alveolar type II epithelial (A549) cells and Klebsiella pneumoniae, an in vitro model of K. pneumoniae-infected A549 cells was established. Western blot analysis and immunofluorescence staining were used to detect the distribution of microtubule-associated protein 1A/1B-light chain 3 (LC3) and the expression of the LC3-phosphatidylethanolamine conjugate (LC3-II). K. pneumoniae-infected A549 cells were treated with different concentrations of an autophagy inhibitor or promoter for different time periods to assess the level of autophagy. Western blot analysis and immunofluorescence staining showed that K. pneumoniae could induce autophagy by A549 cells. With an increase in bacterial concentration and time of infection, autophagy gradually increased. The autophagy inhibitor significantly downregulated, while the promoter upregulated, expression of the autophagy-related protein LC3-II. Autophagy plays an important role in the resistance of alveolar type II epithelial (A549) cells to K. pneumoniae infection.

Cloning, expression, and protective immunity in mice of a gene encoding the diagnostic antigen P-29 of Echinococcus granulosus.

Taeniid tapeworm Echinococcus granulosus is the causative agent of Echinococcosis, an important zoonosis with worldwide distribution. In this study, a diagnostic antigen P-29 was cloned from E. granulosus and expressed in Escherichia coli. Sequence analysis showed that EgP-29 contains 717-bp open reading frame and encodes a protein of 238 amino acid residues with a predicted molecular weight of 27.1 kDa. The recombinant EgP-29 (rEgP-29) could be recognized with antimice sera in Western blotting. The specific antibody was detected by enzyme-linked immunosorbent assay. Mice vaccinated with rEgP-29 and challenged intraperitoneally with E. granulosus protoscoleces revealed significant protective immunity of 96.6% (P<0.05), compared with the control group. Thus, rEgP-29 protein is a promising candidate for an effective vaccine to prevent secondary echinococcosis.

Newborn Screening of Genetic Mutations in Common Deafness Genes With Bloodspot-Based Gene Chip Array.

PURPOSE: This study screens for deafness gene mutations in newborns in the Northwest China population. METHOD: The 9 sites of 4 common deafness genes (GJB2, GJB3, SLC26A4, and mt 12S rRNA) were detected by bloodspot-based gene chip array in 2,500 newborns. RESULTS: We detected mutations of the 4 genes in 101 (4.04%) newborns; particularly, 0.20% detected the double mutations. In the Hui population, 4.58% of the newborns tested positive for mutations, whereas 4.01% of Han newborns tested positive for mutations. The detective rates are as follows: 1.44% for GJB2 235delC, 1.08% for SLC26A4 IVS7-2A>G, 0.48% for GJB2 299_300delAT, 0.28% for SLC26A4 2168A>G, 0.2% for mt 12S rRNA 1555A>G, and 0.16% for GJB3 538C>T. The 31.25% (5/16) of infants with GJB2 235delC, 50% (3/6) with GJB2 299_300delAT, and 25% (3/12) with SLC26A4 IVS7-2A>G showed abnormal hearing when tested; only 1 double mutation case received the hearing test, and this infant showed abnormality in both ears on the hearing test. CONCLUSIONS: High mutation rates in the common deafness genes were detected in newborns in Northwest China. Our study is helpful in understanding the deafness genomic epidemiology and also provides evidence for prenatal and postnatal care as well as policy making on population health in the region.

Molecular Characteristics of Carbapenem-Resistant Enterobacter cloacae in Ningxia Province, China.

The emergence of carbapenem-resistant Enterobacteriaceae (CRE) has become a major public health concern worldwide and a new challenge in the treatment of infectious diseases. The molecular characteristics of Enterobacter cloacae in Ningxia China are unknown. In this study, we reported 10 carbapenem-resistant E. cloacae isolates from the General Hospital of Ningxia Medical University, the largest university hospital in Ningxia between January 2012 and December 2013. Bacteria isolates were identified by Vitek2 compact and the identity of non-duplicate E. cloacae isolates was further confirmed by PCR and sequencing. The drug susceptibility and phenotype identification of these isolates were analyzed by agar dilution method, modified Hodge test (MHT), and EDTA synergy test. Beta-lactamase (bla) genes blaNDM-1 was found in 8 out of 10 isolates. Most isolates harbored multiple resistance genes including blaESBL, blaAmpC, quinolones, aminoglycosides, and disinfectant resistance genes. Pulsed field gel electrophoresis (PFGE) showed that these E. cloacae isolates were grouped into 6 clusters based on a cutoff of 80% genetic similarity. In conjugative assay, 9 out of 10 isolates transferred carbapenem-resistant genes to Escherichia coli. Our study has revealed that NDM-1-producing isolates are the most prevalent carbapenem-resistant E. cloacae in Ningxia. These isolates also carry several other carbapenem-resistant genes and can transfer these genes to other bacteria through conjugation. These findings highlight an urgent need to monitor these isolates to prevent their further spread in this region.

Inhibition of MAPK signaling pathways enhances cell death induced by 5-Aminolevulinic acid-photodynamic therapy in skin squamous carcinoma cells.

Combination of a photosensitizer, 5-aminolevulinic acid (ALA), with photodynamic therapy (PDT) has been widely used to treat skin squamous cell carcinoma (SCC). However, a portion of SCC patients do not respond well to PDT. The molecular reason for this resistance is not clear. We hypothesize that mitogen-activated phosphorylation kinase (MAPK) plays a key role in mediating SCC resistance to PDT. To determine whether inhibition of MAPK signaling enhances the anti-tumor effect of ALA-PDT in SCC. The human squamous carcinoma cell line, SCL-1, was either untreated or treated with various combinations of ALA, PDT light source and inhibitors of MAPK signaling components. ALA-PDT treatment significantly decreased cell viability, increased the percentage of annexin-V positive cells and resulted in formation of apoptotic bodies. ALA-PDT treated cells showed increased levels of p-MEK, p-ERK1/2, p-p38, p-Elk-1, p-JNK and p-c-Jun. Addition of inhibitors for ERK1/2 (PD98059), p38 (SB203580) and JNK (SP60125) reversed the changes and led to a more dramatic decrease in SCL-1 cell viability than seen with ALA-PDT alone. Inhibition of the MAPK pathway enhances the cytotoxic effect of ALA-PDT on SCL-1.

The Role of EGFR/ERK/ELK-1 MAP Kinase Pathway in the Underlying Damage to Diabetic Rat Skin.

BACKGROUND: Diabetes mellitus (DM) is a highly prevalent disease. Atrophy and spontaneous ulcers are the most common cutaneous manifestation of diabetic dermopathy (DD). Before spontaneous ulcers, we believe there is an underlying damage stage although the mechanism is unknown. AIMS: To explore the expression of extracellular signal-regulated kinase1/2 (ERK1/2), its correlated upstream protein epidermal growth factor receptor (EGFR) and its downstream transcription factor E twenty-six (ETS)-like 1(ELK-1)in the damage of the diabetic rat skin, and to explore the role of ERK1/2 on the recessive damage to diabetic rat skin. MATERIALS AND METHODS: Eighty Sprague-Dawley (SD) rats weighing 260-300 g were randomly divided into control and streptozotocin (STZ)-induced diabetes groups. After 0.5, 2, 4, and 8 weeks, the shaved skin specimens from the back of rats in both groups were collected to observe the histological characteristics of the skin, to measure the thickness of the epidermis and the dermis, and to observe the ultrastructure. Immunohistochemistry (IHC) and Western blot techniques were used to detect the expression and activation of ERK1/2, EGFR, ELK-1 in the skin of the rats. RESULTS: There are ultrastructural changes in the DM skin. With the continuance of the diabetes course, the thicknesses of the epidermis and dermis decreased, and the expression of phospho-ERK1/2 (P-ERK1/2), EGFR, and ELK-1 was decreased gradually in the back skin of the diabetes rats. It was significantly lower in 4 and 8 week DM than that of the normal control (P < 0.05). The expression of P-EGFR and P-ERK1/2 in the back skin of the diabetes rats was positively correlated (r = 0.572 P < 0.05), and the positive correlation was also obtained between P-ERK1/2 and P-ELK-1 (r = 0.715, P < 0.05). CONCLUSION: THE PHENOMENON OF RECESSIVE DAMAGE EXISTS IN THE SKIN OF DIABETES RATS, WHICH PROBABLY MAY RELATE TO THE WEAKNESS OF THE SIGNAL TRANSDUCTION: P-EGFR → ERK1/2 → ELK-1.

Correlations between digit ratio and infertility in Chinese men.

BACKGROUND: Digit ratio, especially second to fourth digit ratio (2D:4D) is lower in men than in women. The ratios of digit may be established in utero and is negatively correlated with sperm counts and testosterone in men and positively correlated with estrogen in men and women. AIM: To study whether the digit ratio (especially 2D:4D) are associated with the a+b ratio of sperm number in Chinese populations. METHODS: Photocopies of the two hands of 268 men (controls: 72; patients: 196) and seminal parameters of masturbatory semen samples were collected. RESULTS: The mean values of digit ratio of the controls and the patients all presented a trend as 2D:3D<2D:4D<3D:4D<2D:5D<4D:5D<3D:5D; the patients have higher mean values than controls; significant variances of 2D:3D, 3D:4D (left: P<0.05; right: P ≤ 0.05) and 2D:4D (left: P<0.001; right: P<0.01) were found between two groups; there was a higher percentage of 2D>4D in the patients; the relationship between 2D:4D and a+b ratio of sperm number in the patients was significant (P<0.001). CONCLUSION: Digit ratio, especially 2D:4D in the left hand maybe one of the important markers of infertility in men for early diagnosis.