[Evaluation of the efficacy of GnRH-a pretreatment before total hysterectomy for adenomyosis patients: a propensity score matching analysis].

Objective: To evaluate the efficacy of gonadotropin-releasing hormone agonist (GnRH-a) pretreatment before total hysterectomy for adenomyosis patients with uterine volume ≥12 gestational weeks and moderate or severe anemia. Methods: From January 2018 to March 2023, 689 patients who underwent total hysterectomy for adenomyosis in the First Affiliated Hospital of Zhengzhou University were retrospectively analyzed. According to the preoperative medication, they were divided into study group (127 cases) and control group (562 cases). Patients in the study group underwent GnRH-a pretreatment for 3 cycles before surgery, and the control group received operation directly. SPSS 26.0 software was used to perform 1∶1 matching for the two groups of patients through the propensity score matching method. Matching variables included age, body mass index, gravidity, parity, history of pelvic and abdominal surgery, menstrual cycle, menstrual period, dysmenorrhea score, initial diagnosis of cancer antigen 125 (CA125), uterine volume and hemoglobin value. The dysmenorrhea score, uterine volume, hemoglobin value and CA125 level before and after GnRH-a pretreatment in the study group were compared. And the duration of operation, intraoperative blood loss, postoperative white blood cell count, perioperative blood transfusion cases, postoperative disease rate, duration of hospitalization, total hospitalization cost between the two groups were compared. Results: With propensity score matching, 119 patients in the study group and 119 patients in the control group were finally enrolled in this study. In the study group, before and after the treatment with GnRH-a, the dysmenorrhea score (7.4±1.7 vs 5.6±1.8), uterine volume [(362±160) vs (233±126) cm3], hemoglobin value [(74.1±10.7) vs (102.5±13.5) g/L], and CA125 level [(104±76) vs (64±51) kU/L] were statistically different (all P<0.05). There were statistical differences of operation time [(86±18) vs (116±31) minutes], intraoperative blood loss [(24±9) vs (43±22) ml], white blood cell count after 1 day of operation [(9.80±0.10)×109/L vs (9.90±0.10)×109/L], number of perioperative blood transfusion case [5.9% (7/119) vs 61.3% (73/119)], postoperative disease rate [5.0% (6/119) vs 16.0% (19/119)], hospitalization duration [(7.1±1.6) vs (7.9±1.6) days], and total hospitalization cost [(35 323±5 275) vs (37 159±5 640) yuan] between the study group and the control group (all P<0.05). Conclusion: The pretreatment of using GnRH-a before total hysterectomy for adenomyosis patients with uterine volume ≥12 gestational weeks and moderate or severe anemia is not only conducive to improving dysmenorrhea, signs of anemia, reducing uterine volume, but also conducive to the implementation of surgery, reducing intraoperative and postoperative complications, and reducing hospital costs.

Segmental mandibular advancement for moderate-to-severe obstructive sleep apnoea: a pilot study.

Segmental mandibular advancement (SMA) consists of a combination of bilateral sagittal split osteotomy, anterior subapical osteotomy with extraction of the first premolars, and genioplasty, to allow an extended advancement of the mandible for the improvement of tongue base obstruction in moderate-to-severe obstructive sleep apnoea (OSA) and to minimize any unfavourable aesthetic change due to the large jaw advancement. The aim of this pilot study was to evaluate the surgical outcomes and complications following SMA in OSA patients. Twelve patients (nine male, three female) underwent SMA as part or whole of their skeletal advancement procedure for OSA. The apnoea-hypopnoea index (AHI) improved from a mean± standard deviation 42.4 ± 22.0/hour preoperatively to 9.0 ± 17.4/hour at 1 year postoperative. Surgical success (50% reduction in AHI) was achieved in 11 of the 12 patients (91.7%) at 1 year postoperative, while seven patients (58.3%) attained surgical cure (AHI<5/hour). The lowest oxygen saturation (LSAT) increased from a mean 73.3% preoperatively to 78.7% at 1 year postoperative. The airway volume increased from a mean 2.4 ± 1.7 cm3 at baseline to 6.7 ± 3.5 cm3 at 1 year postoperative (P < 0.001). No major complication occurred. This pilot study showed that SMA appears to be safe and effective as part or whole of the skeletal advancement surgery for moderate-to-severe OSA.

[Impact of probiotics on the lung development of Bama minipig after premature birth].

Objectives: To examine the impact of probiotics on the lung development of preterm birth of Bama pig. Methods: From April 2020 to October 2021, this animal experimental research was performed by setting up preterm (birth at gestation 104 d), full-term (birth at gestation 113 d), preterm with probiotics (birth at gestation 104 d treated with probiotics given at 3 d after birth), and full-term with probiotics (birth at gestation 113 d treated with probiotics given at 3 d after birth) groups and using the preterm Bama minipig model, the body weights were recorded and lung, ileum, and intestinal content samples were collected at birth, 4 days, 9 days, and 21 days after births of the piglets in preterm and full-term groups, the same samples were collected on 9 days after births of the piglets in preterm with probiotics and full-term with probiotics groups. The body weight and radial alveolar counts (RAC) were compared to evaluate the lung development of the piglets. The lengths of ileal villus were compared to evaluate the development of ileum. The composition structures of bacteria in ileum were analyzed by 16 S rRNA sequencing. The statistical analyses between different groups were performed by t test. Results: There were totally 30 piglets (16 female piglets and 14 male piglets) involving 12 piglets in preterm and full-term groups respectively and 3 piglets in preterm with probiotics and full-term with probiotics groups respectively. The body weights of the piglets in preterm group were lower than those in full-term group at 4, 9 and 21 d after birth ((507±27) vs. (694±56) g, (620±35) vs. (1 092±154) g, (1 660±210) vs. (2 960±418) g,t=2.96, 2.99, 2.78, all P<0.05). The alveolarization of the preterm piglets at 9 days after birth was significantly lower than that of the full-term piglets at the equivalent time point (4.00±0.29 vs. 6.11±0.35, t=4.64, P<0.01). The bacteria genus with the highest abundance in ileum were all different between the preterm and the full-term groups at 4, 9 and 21 d after birth (4 d Escherichia-Shigella (26.63%) and Enterococcus (30.48%) respectively;9 d Turicibacter (35.94%) and Lactobacillus (27.33%) respectively;21 d Escherichia-Shigella (28.02%) and Lactobacillus (46.29%) respectively). The heights of ileal villus of the preterm piglets at 9 d after birth were significantly lower than those of the full-term minipigs at the equivalent time point ((297±21) vs. (411±32) µm, t=3.01, P=0.007).There were both no differences in the body weight and alveolarization ((692±36) vs. (767±67) g, 5.44±0.34 vs. 5.89±0.26, t=0.74, 1.04, both P>0.05) between the piglets in preterm with probiotics group and those in full-term with probiotics group. Turicibacter was the dominant genus in the piglets of both preterm with probiotics and the full-term with probiotics groups. The heights of ileal villus of the piglets in preterm with probiotics group were significantly longer that those of the piglets in preterm group ((371±13) vs. (297±21) µm, t=3.04, P=0.006), and were both not significantly different from those of the piglets in full-term with probiotics group and full-term group ((371±13) vs. (338±12) and (411±32) µm, t=1.90, 1.15, both P>0.05). Conclusions: Premature birth could impact the lung alveolarization of piglets. The probiotics could improve the lung alveolarization of preterm minipigs by promoting the development of ileum.

Time series analysis and forecasting of chlamydia trachomatis incidence using surveillance data from 2008 to 2019 in Shenzhen, China.

Chlamydia trachomatis (CT) infection has been a major public health threat globally. Monitoring and prediction of CT epidemic status and trends are important for programme planning, allocating resources and assessing impact; however, such activities are limited in China. In this study, we aimed to apply a seasonal autoregressive integrated moving average (SARIMA) model to predict the incidence of CT infection in Shenzhen city, China. The monthly incidence of CT between January 2008 and June 2019 in Shenzhen was used to fit and validate the SARIMA model. A seasonal fluctuation and a slightly increasing pattern of a long-term trend were revealed in the time series of CT incidence. The monthly CT incidence ranged from 4.80/100 000 to 21.56/100 000. The mean absolute percentage error value of the optimal model was 8.08%. The SARIMA model could be applied to effectively predict the short-term CT incidence in Shenzhen and provide support for the development of interventions for disease control and prevention.

MicroRNA-100 inhibits bone morphogenetic protein-induced osteoblast differentiation by targeting Smad1.

OBJECTIVE: MicroRNAs (miRNAs) act as key regulators of diverse cellular activities by regulating the expression of protein-coding genes. Osteoblast differentiation, a fundamental step in skeletal development, involves the activation of several signaling pathways, including transforming growth factor ß (TGF-ß), bone morphogenetic protein (BMP), and Wnt signaling pathways. MATERIALS AND METHODS: miRNA expression was measured using TaqManRT-PCR. Western blot was used to detect the protein expression of Smad1. Luciferase reporter assay was used to measure the luciferase activity. RESULTS: In this study, we found that miR-100 was expressed in mesenchymal progenitor cell lines; furthermore, its expression was reduced during osteoblast differentiation. Retroviral overexpression of miR-100 decreased Smad1 protein levels, whereas miR-100 inhibition had the opposite effect, suggesting that miR-100 acts as an endogenous attenuator of Smad1 in osteoblast differentiation. CONCLUSIONS: Together, our data demonstrate that miR-100 acts as an important endogenous negative regulator of BMP-induced osteoblast differentiation.

Silencing of NUF2 inhibits proliferation of human osteosarcoma Saos-2 cells.

OBJECTIVE: NUF2 (NUF2, Ndc80 kinetochore complex component), which is essential for kinetochore-microtubule attachment in mitosis, has emerged as a critical mediator of the cell cycle in multiple tumour occurrences. In the present study, we aimed to investigate the role of NUF2 in osteosarcoma, one of the most common primary bone tumours in children and young adults. MATERIALS AND METHODS: Lentivirus-mediated short-hairpin RNA (shRNA) targeting NUF2 (Lv-shNUF2) was employed for evaluation in human osteosarcoma Saos-2 cells. After NUF2 silencing, the proliferation of Saos-2 cells was significantly inhibited, as determined by the MTT assay. RESULTS: The colony forming ability was also significantly decreased in Saos-2 cells infected with Lv-shNUF2. Flow cytometry revealed that downregulation of NUF2 in Saos-2 cells caused a remarkable accumulation of the cell population in the S phase. Furthermore, the expression levels of cell cycle regulators cyclin A and cyclin-dependent kinase 2 (CDK2) were notably decreased, whereas those of cyclin-dependent kinase inhibitors p21Cip1 and p27Kip1, were increased in response to NUF2 knockdown in Saos-2 cells. CONCLUSIONS: Our findings suggest that NUF2 might modulate cell proliferation via cell cycle control in Saos-2 cells. Downregulation of NUF2 by shRNA might be a novel strategy for early treatment of osteosarcoma using molecular-targeting therapy.

Molecularly imprinted polymer doped with Hectorite for selective recognition of sinomenine hydrochloride.

In this work, a new and facile method was introduced to prepare molecularly imprinted polymers (MIPs) based on nano clay hectorite (Hec) for sinomenine hydrochloride (SM) analysis. Hec was firstly dissolved in distilled water in order to swell adequately, followed by a common precipitation polymerization with SM as the template, methacrylic acid as monomer, ethylene glycol dimethacrylate as a crosslinker and 2,2-azobisisobutyronitrile as an initiator. Hec@SM-MIPs were characterized by Fourier transform infrared spectrometer, transmission electron microscopy, scanning electron microscopy, energy dispersive X-ray spectroscopy and X-ray diffraction. The maximum binding capacity of Hec@SM-MIPs, SM-MIPs and non-imprinted polymers (NIPs) (Hec@NIPs) was 57.4, 16.8 and 11.6 mg/g, respectively. The reason for this result may be that Hec@SM-MIPs have more binding sites and imprinted cavities for template molecule. Equilibrium data were described by the Langmuir and Freundlich isotherm models. The results showed that the Hec@SM-MIPs adsorption data correlated better with the Langmuir equation than the Freundlich equation under the studied concentration range. In vitro drug release experiment, Hec@SM-MIPs have a better ability to control SM release than SM-MIPs. Therefore, Hec@SM-MIPs were successfully applied to extraction of SM and used as the materials for drug delivery system.

BMP-2 promotes chondrogenesis of rat adipose-derived stem cells by using a lentiviral system.

Osteoporosis poses a major public health threat in aging societies. Adipose-derived stem cells (ADSCs) are multipotent adult stem cells that have the ability to yield mesenchymal stem cells, and have the potential to undergo osteogenesis and bone regeneration. Bone morphogenetic proteins (BMPs) have been demonstrated to upregulate bone gene expression after mechanical injury and to improve bone injury repair. This study aimed to produce BMP-2 expression in ADSCs by using lentiviral vectors. Subcutaneous adipose tissue from 4-week-old male Sprague-Dawley rats was used. Oil red O staining was used to detect adipocyte formation from ADSCs. Induction of ADSC osteogenesis was confirmed with Alizarin red S staining. The recombinant lenti-hBMP-2/neo was constructed to infect ADSCs, BMP-2 expression was measured by immunoblotting analysis, and cellular alkaline phosphatase levels were examined. We found that >70% of ADSC cells could be induced to differentiate into osteocytes or adipocytes. Under osteogenic induction, ADSCs showed increased intracellular calcium deposition, the formation of calcium tubercles, and the disappearance of cellular structures in calcium tubercles. After infection of ADSCs by lenti-hBMP-2/neo, BMP-2 was expressed after doxycycline induction. We, thus, conclude that ADSCs maintain vigorous growth ex vivo and possess stem cell-like properties. When infected with lenti-hBMP-2/neo, ADSCs can be induced to promote BMP-2 expression.

Molecular cloning, sequence analysis, and function of the intestinal epithelial stem cell marker Bmi1 in pig intestinal epithelial cells.

In the present work, we cloned the full-length cDNA of the pig Bmi1 gene (BMI1 polycomb ring finger oncogene), which has been indicated as an intestinal epithelial stem cell (IESC) marker in other mammals. This paper provides the first report of the function of Bmi1 in pig intestinal epithelial cells and a brief description of its underlying mechanism. Rapid amplification of cDNA ends technology was used to clone the complete pig Bmi1 sequence, and a Bmi1-pcDNA3.1 vector was constructed for transfection into an intestinal porcine epithelial cell line (IPEC-1). The proliferation ability of the cells was estimated using the MTT assay and the EdU incorporation method at different time points after seeding. Cell cycle information was detected by flow cytometry. The mRNA abundances of cell cycle-related genes were also measured. The results indicated that the pig Bmi1 cDNA is 3,193 bp in length and consists of a 981 bp open reading frame, a 256 bp 5´ untranslated region (UTR), and a 1,956 bp 3' UTR. The transcript contains no signal peptides, and there are no transmembrane regions in the pig Bmi1 coded protein, which has a total of 326 AA. The overexpression of the pig Bmi1 in the IPEC-1 cells led to increased cell proliferation and a lower percentage of cells in the G1 and S phases (P < 0.05), along with a higher percentage of cells in the G2 phase (P < 0.05). Furthermore, the gene expression levels of PCNA, Cyclin D1, Cyclin D2, Cyclin B, CDK1, and CDK2 were all elevated (P < 0.05) by Bmi1 overexpression, while the gene expression levels of Cyclin A2 and p21 showed little difference (P > 0.05). Our data suggested that pig Bmi1 can increase the proliferation of IPEC-1 cells by promoting the G1/S transition and the overall cell cycle process.

Ultrastructural localization of laminin and type IV collagen in normal human breast.

The ability of carcinomas to invade and metastasize depends in part on their passage through basement membranes. Two of the major components of basement membranes are type IV collagen and laminin and these have been extensively studied by light microscopical immunocytochemical techniques to investigate alterations in their distribution in human breast carcinomas [1-3]. Breaks in the epithelial basement membrane associated with malignant epithelial tumors have been reported [3, 4]. However, these breaks are based on immunocytochemical observations and have not been correlated with basement membrane morphology. By light microscopical techniques there is no evidence of type IV collagen or laminin between myoepithelial cells or between myoepithelial and epithelial cells and they appear to be restricted to the basement membrane surrounding the entire ductule of the breast. By electron microscopy the basement membrane region contains a linear, homogeneous, electron-dense region (lamina densa) beneath a clear zone (lamina lucida) directly beneath the epithelial and myoepithelial cells of the breast ductule. These two regions constitute the basal lamina. Hemidesmosomes are present on the basal aspect of myoepithelial cells where fine filaments connect these regions to the adjacent basal lamina. The extracellular matrix components laminin and type IV collagen have both been localized in the basement membrane of the normal human breast ductule. Breaks in the continuity of these components occur in breast carcinomas and have been implicated in tumor metastasis. Using a postembedding ultrastructural immunogold technique, laminin and type IV collagen were distributed within the basal lamina surrounding the normal human breast ductule. Both components were present diffusely along the basal lamina and were not localized to particular regions, and neither were present between epithelial and myoepithelial cells. Laminin binding of these cells thus probably occurs only at the basal aspect where they are in contact with the basal lamina and is not involved in the cell-cell adhesion between epithelial and myoepithelial cells. This study provides a basis for further ultrastructural investigations of extracellular matrix components in normal and neoplastic breast tissue.

Antigenic specificity of anti-neutrophil cytoplasmic antibody.

PURPOSE: To determine the antigens recognized by sera containing classic anti-neutrophil cytoplasmic antibodies (c-ANCAs) and perinuclear anti-neutrophil cytoplasmic antibodies (p-ANCAs). METHODS: A total of 160 serum samples (all from a reference laboratory) that were originally collected from different clinics for ANCA tests were examined for c-ANCA and p-ANCA by indirect immunofluorescence (IIF). All positive sera were further tested for reactivity to proteinase 3 (PR3), myeloperoxidase (MPO), lactoferrin (LF), and lysozyme (LZ) by enzyme-linked immunosorbent assay (ELISA). In addition, sera from 110 patients with systemic lupus erythematosus (SLE), 51 patients with rheumatoid arthritis (RA), and 40 healthy subjects were also tested for reactivity to these antigens. RESULTS: HF detected ANCA in 81 (51%) of the 160 clinical serum samples. Of these 81 serum samples, 21 (26%) contained c-ANCA and 60 (74%) contained p-ANCA. P-ANCA was more commonly found in antinuclear antibody (ANA)-positive sera than in ANA-negative sera (p < 0.01). Of the 21 serum samples positive for c-ANCA, 12 (57%) reacted to PR3, four (19%) to LF, four (19%) to LZ, and three (14%) to MPO on ELISA. By contrast, of the 60 sera positive for p-ANCA, 15 (25%) reacted to MPO, 13 (22%) to PR3, eight (13%) to LF, and four (7%) to LZ. The prevalence of ANCA specificities in serum samples from SLE patients were as follows: anti-PR3, 0%; anti-MPO, 1%; anti-LF, 27%; and anti-LZ, 29%. The prevalence of ANCA specificities in serum samples from RA patients were as follows: anti-PR3, 6%; anti-MPO, 2%; anti-LF, 8%; anti-LZ, 4%. CONCLUSION: Sera positive for c-ANCA and p-ANCA reacted to diverse cytoplasmic antigens from neutrophils. P-ANCA was found in 55% of ANA-positive serum samples. LF and LZ were most commonly found in serum samples from patients with SLE.

Synthesis and expression of a gene from kringle-2 domain of tissue plasminogen activator in E. coli.

The DNA fragment corresponding to the tissue plasminogen activator (tPA) sequence 174-262 (Kringle-2 domain) has been synthesized by using the solid phase phosphotriester method. The Kringle-2 domain of human tPA was expressed in Escherichia coli by secretion into the periplasmic space using the Lpp-Lac promoter and PIN-III OmpA2 signal sequence. About two thirds of the expression product was secreted into the periplasmic space, and purified with ammonium sulfate fractionation, affinity chromatography on Lysine-Sepharose, and FPLC-Mono Q exchange chromatography. The amino acid composition observed from the Kringle-2 purified from E. coli is identical with that expected for the 174-262 fragment of human tPA. Radio binding assay shows that the recombinant Kringle-2 domain possesses the activity of fibrin binding.

Expression of Epstein-Barr virus-encoded proteins in nasopharyngeal carcinoma.

Expression of the Epstein-Barr virus (EBV) encoded nuclear antigens (EBNA 1 to 6) and membrane-associated protein (LMP) was investigated by immunoblotting in 83 nasopharyngeal carcinoma (NPC) biopsies and 25 other tumor and normal tissue specimens from the head and neck region. Fifty-eight of the 83 NPC biopsies were large enough to yield parallel data on virus DNA and viral expression. All 16 cases of clinically diagnosed and histologically confirmed NPCs from North Africa contained EBV DNA and expressed EBNA-1. Of 31 clinically diagnosed NPCs from China, 29 contained EBV DNA and 25 of these expressed EBNA-1. One control tissue biopsy from the oropharynx of NPC patients contained EBV DNA, but none expressed EBNA-1. The latent membrane protein (LMP) was detected in 22/31 of the Chinese and in 10/16 of the North African NPC biopsies. None of the NPC biopsies or control tissues expressed detectable amounts of EBNA 2 or any of the other 4 nuclear antigens which are invariably expressed in EBV-transformed B cells. A smaller number of tumors from Malaysia and East Africa exhibited a similar pattern of expression. EBV was rescued from a nude-mouse-passaged North African NPC tumor by co-cultivation of the tumor cells with umbilical cord blood lymphocytes. The tumor expressed EBNA 1 and LMP, but not EBNA 2 or the other 4 EBNAs. The resulting LCLs expressed all 6 nuclear antigens, EBNA 1 to 6 and LMP. Our data suggest that expression of the EBV genome is regulated in a tissue-specific fashion.