RACGAP1 knockdown synergizes and enhances the effects of chemotherapeutics on ovarian cancer.

Among the three most prevalent cancers affecting the female reproductive system, ovarian cancer (OV) ranks as the second most frequently diagnosed. It is important to investigate the genomic complexity of OV to develop diagnostic and therapeutic strategies. Through the utilization of bioinformatics analysis, it was determined that RacGTPase Activating Protein 1 (RACGAP1) holds significant significance in the field of OV chemotherapeutics, an aspect that has not been thoroughly explored in prior investigations. In our study, a notable increase in RACGAP1 expression was detected in ovarian cancer, demonstrating a robust association with clinicopathological features and patient prognosis. In vivo and in vitro testing revealed that RACGAP1 acts synergistically with chemotherapeutics to enhance their effects on ovarian cancer. Furthermore, an interaction between RACGAP1 and the subunit G2 of the condensin II complex, known as non-SMC condensin II complex subunit G2 (NCAPG2), has been identified. Our findings may provide new insight for improving therapeutic strategies for OV.

Type 2 diabetes mellitus and the risk of male infertility: a Mendelian randomization study.

Objective: To assess the causal effect of type 2 diabetes mellitus (T2DM) on male infertility (MI) and erectile dysfunction (ED) by Mendelian randomization (MR) analysis. Methods: Data for T2DM, MI, and ED were obtained from genome-wide association studies (GWAS) involving 298, 957, 73, 479, and 223, 805 Europeans, respectively. We performed univariate MR analysis using MR Egger, Weighted median (WM) and Inverse variance weighted (IVW) methods to assess causal effects among the three. Through the Genotype Tissue Expression (GTEx) database, single-nucleotide polymorphisms (SNPs) that affect the expression levels of T2DM-related genes were located using expression quantitative trait loci (eQTL). Results: MR analysis showed a significant causal relationship between T2DM and ED (WM, OR: 1.180, 95%CI: 1.010-1.378, P = 0.037; IVW, OR: 1.190, 95%CI: 1.084-1.300, P < 0.001). There is also a significant causal relationship between T2DM and MI (MR Egger, OR: 0.549, 95%CI: 0.317-0.952, P = 0.037; WM, OR: 0.593, 95%CI: 0.400, P = 0.010; IVW, OR: 0.767, 95%CI: 0.600-0.980, P = 0.034). ED may not cause MI (P > 0.05). We also found that rs6585827 corresponding to the PLEKHA1 gene associated with T2DM is an eQTL variant affecting the expression of this gene. Conclusion: T2DM has a direct causal effect on ED and MI. The level of PLEKHA1 expression suppressed by rs6585827 is potentially associated with a lower risk of T2DM.

Establishment and validation of haematological reference intervals for newborns aged 5 to 28 days in Nanjing, China.

BACKGROUND: China lacks a standard for reference intervals (RIs) of complete blood cell (CBC) counts in newborns. This study aimed to determine local haematological RIs for newborns. METHODS: This prospective study was conducted from January 2020 to December 2020 in Nanjing Maternity and Child Health Care Hospital. We collected capillary blood specimens from 497 healthy newborns aged 5-28 days. We calculated the RIs as nonparametric 2.5th to 97.5th percentiles and 90% confidence intervals following the EP28-A3c guideline. We validated the RIs in another 20 specimens from healthy newborns. RESULTS: The RIs for the 18 CBC parameters were: white blood cell count 7.17-15.69 × 109/L; monocytes# 0.52-1.66 × 109/L; Mono% 5.7-14.1; eosinophils# 0.16-1.08 × 109/L; Eos% 1.5-8.9; Lymphocytes# 4.04-8.08 × 109/L; Lymph% 40.1-67.8; Neutrophils# 1.59-6.41 × 109/L; Neut% 18.1-46.7; Basophils# 0.00-0.04 × 109/L; Baso% 0.0-0.4; red blood cell count 3.38-5.89 × 1012/L; haemoglobin (Hb) 116-198 g/L; Hematocrit % 35.2-61.2; mean corpuscular Hb (MCH) 30.9-36.3 pg/L; MCH concentration 310-341 g/L; mean corpuscular volume 94.5-112.2 fL; and platelets 210-610 × 109/L. The RIs had a conformity rate of 90%-100% in the validation specimens. CONCLUSION: We established region-specific RIs for CBC parameters in healthy newborns to help diagnose haematological disease in neonates.

[Strategy of acetabular anteversion in total hip arthroplasty with lumbar degenerative kyphosis].

OBJECTIVE: To investigate how to place the anteversion of acetabular prosthesis more reasonably in patients with lumbar degenerative kyphosis. METHODS: A total of 122 patients with degenerative kyphosis of lumbar spine who underwent total hip arthroplasty from December 2017 to October 2019 were included and divided into experimental group and control group, 61 cases in each group. In experimental group, there were 25 males and 36 females, with a median age of 67.0 years;the median course of disease was 46.0 months;the functional pelvic plane with acetabular anteversion was set according to different types of pelvic anterior plane bracket. In control group, there were 27 males and 34 females, with a median age of 67.0 years;the median course was 42.0 months;in control group, the anteversion was set by the traditional method. The patients were followed up for 3 months. The operation time and blood loss were recorded. The incidence of infection and dislocation within 3 months was counted. Harris score before and 3 months after operation was recorded. Functional anteversion angle of standing position was measured 3 months after operation. RESULTS: Compared with control group, there was no difference in operation time and blood loss between the two groups (P=0.918, 0.381);there was no infection between two groups within 3 months after operation;there was 1 case of hip joint dislocation in the control group and no dislocation in experimental group. There was no significant difference in Harris score before and after operation. Three months later, reexamination of pelvic standing radiographs showed that the number of patients with functional anteversion of acetabular prosthesis outside the safe area was less in experimental group thanin control group (P=0.048), and the functional anteversion angle of acetabular prosthesis was more concentrated in the range of 15 ° to 20 ° in experimental group (P<0.001). CONCLUSION: According to the preoperative evaluation and classification of patients, better functional anteversion of acetabular prosthesis can be obtained with the help of pelvic anterior plane reference bracket in hip arthroplasty with lumbar degenerative kyphosis.

[Anatomical and tensile mechanical analysis of hip joint capsule repair in total hip replacement].

OBJECTIVE: To explore the tensile mechanics and anatomical characteristics of the posterior hip capsule, and provide biomechanical and anatomical evidence for capsule repair in total hip replacement. METHODS: Six bone-capsule-bone specimens were obtained from posterior hip joint of fresh frozen cadavers. The maximum strain, load, elastic modulus and load strain curves of the capsule ligament complex specimens were recorded by Instron Universal Material Testing Machine. Twelve cadaveric hip specimens were dissected to the capsule. The tensile strain of normal capsule and conventionally reconstructed capsule at 90 degrees of hip flexion were documented. The suture area of the posterior capsule was divided into nine sections, and the thicknessof different sections was measured and compared. Posterior capsule of the cadavers was repaired in conventionally way and anatomical way separately and simulated rehabilitation was conducted. The effect of rehabilitation on the repaired capsule was observed. RESULTS: The load-strain curve of capsule ligament complex conforms to rheological and viscoelastic characteristics. The maximum tensile strain of the complex was (39.21±5.23)%, the maximum load was (142.06± 34.15) N, the tensile strength was (1.65±0.38) MPa, and the elastic modulus is (14.23±5.62) MPa. At 90 ° hip flexion, the tensile strain of repaired capsule was higher than that of normal capsule, and the difference was statistically significant (P< 0.05). Tensile strain of conventionally reconstructed capsule is:upper part (37.0±4.9)%, middle part ( 53.3±1.1)%, lower part (68.3±6.2)%, tensile strain of normal capsule is:upper part (17.0±2.6)%, middle part (24.1±1.4)%, lower part (26.0± 4.3)% . The thickness of the posterior joint capsulein different sections is statistically significant (P<0.05), and capsule at 0.5cm proximal to the femoral insertion is suitable for suture. There the average thickness of capsule is:upper part (3.48 ± 0.11) mm, middle part (2.36 ± 0.09) mm, lower part (1. 59±0.24) mm. The posterior inferior joint capsule is thinnest at (1.42± 0.02) cm proximal to the femoral insertion, and sutures should be avoided here. After simulating rehabilitation, avulsion occurred in the lower part of the posterior capsule repaired conventionally (10/12), and the anatomically repaired capsule remained intact. CONCLUSION: The lower part of conventionally repaired capsule is overstretched and tends to fail. Anatomically repaired capsule conforms to tensile mechanics and is helpful to reduce the failure rate of repair.

Sperm DNA fragmentation in Chinese couples with unexplained recurrent pregnancy loss.

We aimed to study the association between sperm DNA fragmentation and recurrent pregnancy loss (RPL) in the Chinese population via a retrospective observational study of Chinese couples who had experienced RPL between May 2013 and August 2018. The study population included 461 men from couples with RPL and 411 men from a control group (couples with clinical pregnancy via in vitro fertilization owing to female causes). Routine semen analysis, sperm chromatin analysis, and microscopic (high-power) morphological analysis were performed using semen samples. Semen samples were assessed for volume, sperm count, and motility. The sperm DNA fragmentation index (DFI) was calculated, and the median DFI was obtained. Men were categorized as having normal (37.8%; DFI ≤ 15.0%), moderate (33.6%; 15.0% < DFI < 30.0%), or severe (28.6%; DFI ≥ 30.0%) DNA fragmentation levels. The percentage of men with severe DNA fragmentation was significantly higher in the RPL (42.3%) group than that in the control group (13.1%), whereas the percentage of men with normal levels of DNA fragmentation was significantly lower in the RPL group (22.8%) than that in the control group (54.7%). Subsequent analysis also demonstrated that the sperm DNA fragmentation rate had a moderate reverse correlation with the sperm progressive motility rate (r = -0.47, P < 0.001) and the total motile sperm count (r = -0.31, P < 0.001). We found a positive correlation between RPL and sperm DNA fragmentation. The results suggest that increased sperm DNA damage is associated with RPL.

Intra-articular, single-shot co-injection of hyaluronic acid and corticosteroids in knee osteoarthritis: A randomized controlled trial.

The aim of the present study was to investigate whether the co-injection of hyaluronic acid (HA) and corticosteroids (CS) was superior to HA alone in the treatment of knee OA. A total of 120 participants with symptomatic knee OA were recruited and formed the intention-to-treat population for a 6-month follow-up. In the HA group, patients received a single-shot injection of 4 ml HA. In the HA&CS group, patients received a co-injection of 3 ml compound betamethasone solution and 4 ml HA. Visual analog scale (VAS), Western Ontario and McMaster University Osteoarthritis Index (WOMAC) and knee flexion motion were assessed as primary outcomes. Patients in the HA&CS group exhibited better pain relief and knee function at the time points of week 1, month 1 and month 3 (P<0.05). For the last follow-up at month 6, the values did not differ significantly between these two groups. Patients in both groups exhibited improvement in pain, knee function, and range of motion following injection. For the final follow-up at month 6, the mean VAS score, WOMAC score and knee flexion motion were still superior to that prior to treatment, but the values did not differ significantly. The co-injection of HA and CS provided a rapid improvement in pain relief, knee function, and range of motion, but did not differ significantly from that of HA alone in the long term effect.

Is exclusion of leukocytes from platelet-rich plasma (PRP) a better choice for early intervertebral disc regeneration?

BACKGROUND: Platelet-rich plasma (PRP) is becoming a promising strategy to treat early intervertebral disc degeneration (IDD) in clinics. Pure PRP without leukocytes (P-PRP) may decrease the catabolic and inflammatory changes in the early degenerated intervertebral discs. The aim of this study was to investigate the effects of P-PRP on nucleus pulposus-derived stem cells (NPSCs) isolated from early degenerated intervertebral discs in vitro. METHODS: NPSCs isolated from early degenerated discs of rabbits were treated with P-PRP or leukocyte-platelet-rich PRP (L-PRP) in vitro, followed by measuring cell proliferation, stem cell marker expression, inflammatory gene expression, and anabolic and catabolic protein expression by immunostaining, quantitative real-time polymerase chain reaction, Western blot, and enzyme-linked immunosorbent assay. RESULTS: Cell proliferation was induced by P-PRP in a dose-dependent manner with maximum proliferation at 10% P-PRP dose. P-PRP induced differentiation of NPSCs into active nucleus pulposus cells. P-PRP mainly increased the expression of anabolic genes and relative proteins, aggrecan (AGC), collagen types II (Col II), while L-PRP predominantly increased the expression of catabolic and inflammatory genes, matrix metalloproteinase-1 (MMP-1), MMP-13, interleukin-1 beta (IL-1ß), IL-6, tumor necrosis factor alpha (TNF-α), and protein production of IL-1ß and TNF-α. CONCLUSIONS: Leukocytes in PRP activate inflammatory and catabolic effects on NPSCs from early degenerated intervertebral discs. Hence, P-PRP may be a more suitable therapeutic strategy for early IDD.

Treatment for Periprosthetic Cyst after Total Hip Arthroplasty: Analysis of Six Cases.

The present study investigates the pathogenesis of periprosthetic cysts after total hip replacement, and explores appropriate treatment appoaches. Six patients with periprosthetic cysts after total hip arthroplasty were treated at the First Affiliated Hospital of Nanjing Medical University between 2009 and 2014. During surgery, it was found that all cysts communicated with the hip and the hip prosthesis could be seen after cyst excision. Four patients simply underwent cyst excision, and light red liquid was found in the cyst. Among them, radiological examination revealed that a part of the hip prosthesis projected from the bone bed in one case. Postoperative pathology revealed a synovial cyst with inflammatory cell infiltration. Prostheses were loosened in two cases, so cystectomy and revision of the prosthesis were performed at the same time. Among the six patients, polyethylene wear particles could be seen in five patients through a pathological polarizing microscope. Out of the four patients who underwent simple cyst excision, two patients experienced cyst recurrence within 1 year after surgery; however, there was no cyst recurrence in the two patients who underwent cyst excision and revision of the prosthesis. The formation of a periprosthetic cyst after hip replacement is likely to be related to polyethylene wear and undesirable prosthesis position; in addition, when treated by simple cyst excision, the rate of recurrence was higher.

[Y chromosome microdeletions: detection in 1 052 infertile men and analysis of 14 of their families].

OBJECTIVE: To investigate the characteristics of father-to-son vertical transmission of Y chromosome microdeletions METHODS: We detected the Y by detection of Y chromosome microdeletions in infertile men and analysis of some of their families. chromosome azoospermia factor (AZF) microdeletions in the peripheral blood of 1 052 infertile males, investigated the paternal relatives of 12 cases of AZFc, 1 case of AZFb and 1 case of AZFb + c microdeletions, and drew the family tree diagrams of the infertile paternal relatives according to the findings. RESULTS: Among the 1 052 infertile patients, 89 (9.73%) were found with Y chromosomal microdeletions, including 56 with AZFc, 6 with AZFa, 5 with AZFb, 14 with AZFb + c, and 8 with AZFa + b + c deletion. The investigation of the 14 patients'families revealed 1 case of AZFb and 1 case of AZFb + c deletion de novo. Among the 12 cases of AZFc deletion, vertical heredity was found in 5 patients with severe oligozoospermia, but not in the other 7 with azoospermia. CONCLUSION: AZFe deletion may be vertically inherited from the father in severe oligozoospermia patients, and it is different from the paternal phenotype, while in azoospermia patients, AZF deletion, whatever type it may be, is less likely to be associated with vertical paternal heredity.

[Comparative anatomical study on subinguinal and ilioinguinal approaches and clinical application in acetabular fracture].

OBJECTIVE: To compare anatomical differences between subinguinal and ilioinguinal approaches for the treatment of acetabular fracture and investigate clinical therapeutic effect of subinguinal approach. METHODS: Seven fresh human bodies were chosen, comparative study were performed on the right and left side on the same specimen. Ilioinguinal approaches were adopted on the left and subinguinal were adopted on the right. Inner part of incision started to sun wild above pubic symphysis at 2 cm, and lateral incision ranged from iliac to anterior superior spine about 5 cm. Length and transverse diameter of the first window exposed and lliopsoas freeness were tested and compared. Fifteen acetabular fracture patients treated through subinguinal approach were compared from May 2010 to August 2012. Among all patients, including 12 males and 3 females aged from 20 to 65 years old with an average of 40.6 years old. Matta criteria were used to evaluate clinical outcomes. RESULTS: Length and transverse diameter of the first window exposed and lliopsoas freeness through subinguinal approach were better than through ilioinguinal approach (P<0.01). In 15 patients with acetabular fracture, 10 patients obtained anatomical reduction and 10 patients got satisfied reduction in accordance with Matta criteria. X-ray results of all patients were excellent. CONCLUSION: Compared with ilioinguinal approach, subinguinal approach could enlarge visualization of the first window and simplify surgical procedure. It is an ideal approach to expose anterior and anterior-medialis wall of acetabulum and anterior hip capsule.

[Multi-ligaments reconstruction with single tendon with for the treatment of acute joint dislocation of the first carpometacarpal].

OBJECTIVE: To explore the clinical outcomes of multi-ligaments reconstruction with single tendon in treating acute joint dislocation of the first carpometacarpal. METHODS: From December 2008 to October 2012, 4 patients with acute dislocation of the first carpometacarpal were treated with single carpi radialis longus tendon to reconstruct periarticular four ligaments, which included dorsal ligament, palmar ligament, dorsal radiocarpal ligament, and intermetacarpal ligament between the first and second. There were 3 males and 1 female aged from 22 to 63 years old with an average of 38.7. X-ray, JAMAR grid strength testing, range of motion of carpometacarpal joint and VAS score were used to evaluate clinical outcomes. RESULTS: All patients were followed up from 6 to 40 months with an average of 19 months. The wound were healed well at stage I. No dislocation of the first carpometacarpal joint and signs of joints degeneration occurred on X-ray at 1, 3 and 6 months after operation. JAMAR grip strength recovered from 60 percent to 90 percent of health wide. The results of ROM showed 3 cases recovered flexion and extension of joints and 1 case limited adduction. VAS score was 0 to 1. CONCLUSION: For acute dislocation of the first carpometacarpal joint, reconstruction four ligaments with single carpi radialis longus tendon can be considered the selected treatment,which can restores joint stability and improve joint function.

Antimicrobial and antifouling hydrogels formed in situ from polycarbonate and poly(ethylene glycol) via Michael addition.

A novel class of antimicrobial cationic polycarbonate/PEG hydrogels are designed and synthesized by Michael addition chemistry. These hydrogels demonstrate strong broad-spectrum antimicrobial activities against various clinically isolated multidrug-resistant microbes. Moreover, they exhibit nonfouling properties and prevent the substrate from microbial adhesion. These antimicrobial and antifouling gels are promising materials as catheter coatings and wound dressings to prevent infections.

The role of gC1qR in regulating survival of human papillomavirus 16 oncogene-transfected cervical cancer cells.

Human papillomavirus 16 (HPV-16) is strongly associated with the development of 50% of cervical cancers. The E6 and E7 proteins encoded by high-risk HPV types are associated with the immune evasion of cervical cancer cells, but the mechanism is poorly understood. The purpose of this study was to investigate whether cells transfected with E6 and E7 expression constructs reduce the expression of the globular heads of the C1q receptor (gC1qR), a mitochondrial surface protein overexpressed in certain cancer cells. First, C-33A cells were transiently transfected with the HPV-16 E6 and E7 oncogenes which resulted in gC1qR inhibition and a reduction in apoptosis. Second, gC1qR overexpression in cells showed that caspase-3 activation and mitochondrial dysfunction were involved in gC1qR-induced apoptosis. Cells transfected with a GFP-gC1qR vector resulted in upregulated gC1qR protein and a gradual increase in the generation of reactive oxygen species (ROS). Additionally, ROS generation and increased Ca2+ influx in mitochondria resulted in the loss of the mitochondrial transmembrane potential. Interestingly, when gC1qR was overexpressed in C-33A cells, apoptosis was significantly inhibited when cells were treated with metformin, which may protect mitochondrial function. These data suggest that gC1qR could play an important role in HPV-16-induced cervical cancer immune evasion depending on its level of expression and subcellular localisation.

Gene transfer using self-assembled ternary complexes of cationic magnetic nanoparticles, plasmid DNA and cell-penetrating Tat peptide.

Nonviral magnetofection facilitates gene transfer by using a magnetic field to concentrate magnetic nanoparticle-associated plasmid delivery vectors onto target cells. In light of the well-established effects of the Tat peptide, a cationic cell-penetrating peptide, that enhances the cytoplasmic delivery of a variety of cargos, we tested whether the combined use of magnetofection and Tat-mediated intracellular delivery would improve transfection efficiency. Through electrostatic interaction, gene transfer complexes were generated by mixing polyethylenimine-coated cationic magnetic iron beads with plasmid DNA, followed by addition of a bis(cysteinyl) histidine-rich Tat peptide. These ternary magnetofection complexes provided a 4-fold improvement in transgene expression at a dose of 1 microg of plasmid DNA per 20,000 cells over the binary complexes without the Tat peptide and transfected up to 60% of cells in vitro. The enhanced transfection efficiency was also observed in vivo in the rat spinal cord after lumbar intrathecal injection. Moreover, the injected ternary magnetofection complexes in the cerebrospinal fluid responded to a moving magnetic filed by shifting away from the injection site and mediating transgene expression in a remote region. Thus, our approach could potentially be useful for effective gene therapy treatments of localized diseases.

[Fermentation behaviors of recombinant Pichia pastoris under inhibited methanol concentration].

Chemostat culture was performed to characterize the growth, substrate consumption and the hirudin production, and to disclose their interrelations in the fermentation of recombinant Pichia pastoris. The Andrew substrate-inhibited growth model is more suitable than Monod model to simulate the growth of Pichia pastoris on methanol. Therefore, two stationary states can be obtained in the continuous culture at a certain dilution rate because of the substrate inhibition on cell growth. The stationary state could be obtained if only the dilution rate not more than 0.048 h(-1) in the continuous fermentation. The concentrations of cell, methanol and hirudin were constant after 50 h continuous culture with dilution rate at 0.04 h(-1). However, it could not be obtained when the dilution rate more than 0.048 h(-1) because the other stationary point at S > 0.048 h(-1) is unstable. Therefore, it was found that the cell concentration declined and the methanol concentration increased from 2.9 g/L to 18.1 g/L within 18h at dilution rate 0.06 h(-1). Thus, the fed-batch culture with a constant specific growth rate was carried out to disclose the fermentation behavior at high and constant methanol concentration in aid of a methanol sensor. The theoretical maximum specific growth rate, microm = 0.0464 h(-1), was found under critical methanol concentration, Scrit = 3.1 g/L. The growth of P. pastoris was typically methanol-limited at the methanol concentration S < Scrit. It was, however, inhibited at S > Scrit. The maximum specific Hir65 production rate qp was obtained at 0.2 mg/(g x h) when methanol concentration and mu were 0.5 g/L and 0.02 h(-1), respectively. The specific Hir65 production rate qp increased with the increase of mu and S at mu < 0.02 h(-1), and decreased at mu > 0.02 h(-1). The specific methanol consumption rate increased with the increase of S when S < 5 g/L, but decreased when S > 5 g/L. At last, the high Hir65 production rate 0.2 mg/(g x h) was obtained in the fermentation conducted under methanol-limited concentration and mu controlled at 0.5 g/L and 0.02 h(-1), respectively, while the specific methanol consumption rate is low only at 0.04 g/(g x h), showing the potential for the strategy of getting high Hir65 production rate at the low consumption of methanol.

[Effects of different methanol feeding strategy on hirudin production in high-density fermentation by recombinant Pichia pastoris].

Four different methanol feeding modes were evaluated in the hirudin production in high-density fermentation by Pichia pastoris. It was difficult to avoid methanol excessive in the broth with the feeding strategy only based on DO level. On the other hand, the fluctuation in methanol concentration was observed with methanol feeding strategy by off-line gas chromatography. However, the stable methanol concentration was perfectly achieved by the on-line monitoring with methanol sensor. The supply of energy was improved by feeding glycerol at a limited rate as well as methanol in the induction phase. Therefore, the high cell dry weight (162 g/L) and high hirudin activity (2.4 x 10(4) ATU/mL or 1.7 g/L) was obtained in the fed-batch fermentation of recombinant Pichia pastoris by methanol-glycerol mixed feeding.