The diagnostic value of four-dimensional ultrasound examination of perineum in the diagnosis of postpartum pelvic floor dysfunction

Objective to explore the diagnostic value of perineal four-dimensional ultrasound (4D-US) in postpartum pelvic floor dysfunction (PFD) disease. 328 postpartum PFD patients diagnosed by clinical pelvic floor palpation from June 2018 to December 2020 were selected as the PFD group, and 328 patients without PFD were selected as the control group. All participants underwent perineal 4D-US, the indicators were statistically analyzed. The results showed that the LAT of left and right, LHLR, LHAP, LHA, resting state and holding the breath in the PFD group were higher than those in the control group, and the difference was statistically significant (P<0.05). From cervix to lower margin of pubic symphysis, bladder to lower margin of pubic symphysis of the pubic symphysis, and from the ampulla of the rectum to the lower margin of the pubic symphysis, the PFD group was larger than the control group, but the result of urethral rotation was reversed, and the difference was statistically significant (P<0.05). The morphologic features of the levator ani muscle and pelvic fissure can be detected early using 4D-US, which is a reliable technique that can be learned in a short period of time

The diagnostic value of four-dimensional ultrasound examination of perineum in the diagnosis of postpartum pelvic floor dysfunction

Objective to explore the diagnostic value of perineal four-dimensional ultrasound (4D-US) in postpartum pelvic floor dysfunction (PFD) disease. 328 postpartum PFD patients diagnosed by clinical pelvic floor palpation from June 2018 to December 2020 were selected as the PFD group, and 328 patients without PFD were selected as the control group. All participants underwent perineal 4D-US, the indicators were statistically analyzed. The results showed that the LAT of left and right, LHLR, LHAP, LHA, resting state and holding the breath in the PFD group were higher than those in the control group, and the difference was statistically significant (P<0.05). From cervix to lower margin of pubic symphysis, bladder to lower margin of pubic symphysis of the pubic symphysis, and from the ampulla of the rectum to the lower margin of the pubic symphysis, the PFD group was larger than the control group, but the result of urethral rotation was reversed, and the difference was statistically significant (P<0.05). The morphologic features of the levator ani muscle and pelvic fissure can be detected early using 4D-US, which is a reliable technique that can be learned in a short period of time

Persistence of the Recombinant Genomes of Woodchuck Hepatitis Virus in the Mouse Model

<div><p>Hydrodynamic injection (HI) with a replication competent hepatitis B virus (HBV) genome may lead to transient or prolonged HBV replication in mice. However, the prolonged HBV persistence after HI depends on the specific backbone of the vector carrying HBV genome and the genetic background of the mouse strain. We asked whether a genetically closely related hepadnavirus, woodchuck hepatitis virus (WHV), may maintain the gene expression and replication in the mouse liver after HI. Interestingly, we found that HI of pBS-WHV1.3 containing a 1.3 fold overlength WHV genome in BALB/c mouse led to the long presence of WHV DNA and WHV proteins expression in the mouse liver. Thus, we asked whether WHV genome carrying foreign DNA sequences could maintain the long term gene expression and persistence. For this purpose, the coding region of HBV surface antigen (HBsAg) was inserted into the WHV genome to replace the corresponding region. Three recombinant WHV-HBV genomes were constructed with the replacement with HBsAg a-determinant, major HBsAg, and middle HBsAg. Serum HBsAg, viral DNA, hepatic WHV protein expression, and viral replication intermediates were detected in mice after HI with recombinant genomes. Similarly, the recombinant genomes could persist for a prolonged period of time up to 45 weeks in mice. WHV and recombinant WHV-HBV genomes did not trigger effective antibody and T-cell responses to viral proteins. The ability of recombinant WHV constructs to persist in mice is an interesting aspect for the future investigation and may be explored for <i>in vivo</i> gene transfer.</p></div

Serum HBsAg and intrahepatic WHcAg expression in mice after HI with pWHV-HBV-SS and pWHV-HBV-MS.

<p>(A) Mouse sera were collected after HI with pWHV-HBV-SS (SS) and pWHV-HBV-MS (MS) at the indicated time points and subjected to HBsAg ELISA. SS1-7 and MS1-7 indicated seven individual mice each group. In MS group, two mice of MS4 and MS5 were sacrificed at week 36 after HI, and the mouse splenocytes were taken for ELISpot assay and flow cytometry. The ELISA results were read at OD 450 nm. The cut off value was set as 0.1 and indicated by the dotted lines. (B) Mouse liver samples taken at the indicated time points after HI were detected for WHcAg expression by ICS. A set of representative samples was shown. Magnification: 200×.</p

Detection of the replication intermediates of WHV and the recombinant WHV-HBV genomes in mouse liver.

<p>Mouse liver samples were collected at day 10 after HI with pBS-WHV1.3 (WHV1.3), pWHV-HBV-Sa (Sa), pWHV-HBV-SS (SS), pWHV-HBV-MS (MS) and saline (Mock). The replication intermediates were detected by Southern blot using a full length WHV genome as probe. The signal intensity was quantified by measuring the grey scale using WHV1.3 as the reference. RC DNA: relaxed circular DNA; SS DNA: single stranded DNA.</p

Detection of the surface antigen expression in mouse sera by HBsAg ELISA after HI with pHBsW1-8 and pSaΔP.

<p>Mouse sera were collected at the indicated time points after HI and subjected to HBsAg ELISA. (A) Each group with three mice was hydrodynamically injected with pHBsW1-8 (W1-W8). Mice injected with pHBsBK (pHBs) and saline (mock) were used as positive and negative control, respectively. (B) Eight mice (SaP1-8) were hydrodynamically injected with the mutated pWHV-HBV-Sa, pSaΔP. The results were read at OD 450 nm. The cut off value was set as 0.1 and indicated by the dotted lines.</p

Constructions of recombinant WHV-HBV genomes and transient transfection of recombinant WHV-HBV genomes in hepatoma cells.

<p>(A) The schematic map of recombinant WHV-HBV genomes. pBS-WHV1.3 contained a 1.3 fold overlength WHV genome in pBluescript II SK(+) vector and was used as backbone. The respective WHV genome regions were replaced by the corresponding HBV sequences, shown as black bars. The plasmids pWHV-HBV-Sa, pWHV-HBV-SS, and pWHV-HBV-MS contained HBV sequences encoding HBsAg a-determinant only, major HBsAg and middle HBsAg, respectively. (B) HBsAg expression in the supernatant of the transfected hepatoma cells. Huh7 and HepG2 cells were transiently transfected with plasmids of pBS-WHV1.3 (W1.3), pWHV-HBV-Sa (Sa), pWHV-HBV-SS (SS), pWHV-HBV-MS (MS), pBS-HBV1.3 (H1.3), and pHBc (HBc). The culture supernatants were collected at 24, 48, 72, and 96 hours after transfection for the detection of HBsAg by ELISA. The results were read at OD 450 nm. The cut off value was set as 0.1 and indicated by the dotted line.</p

Serum viral DNA and the expression of viral proteins in mice after HI with pWHV-HBV-Sa.

<p>(A) Six mice (Sa1-Sa6) were hydrodynamically injected with pWHV-HBV-Sa, while four mice (H1.3–1 to H1.3–4) were hydrodynamically injected with pBS-HBV1.3. Mouse sera were collected at the indicated time points after HI and subjected to HBsAg ELISA. The results were read at OD 450 nm. The cut off value was set as 0.1 and indicated by the dotted line. (B) Detection of serum encapsidated viral DNA by PCR. PCR products were visualized by agarose gel electrophoresis. Neg.: Sera from two mice (N1, 2) challenged by saline. (C) Detection of hepatic WHcAg expression by ICS at days 0, 10, 42 and 98 after HI with pWHV-HBV-Sa. A set of representative samples was shown. Magnification: 200×.</p

Serum WHV DNA and intrahepatic viral gene expression after HI with pBS-WHV1.3.

<p>Nine BLAB/c mice (W1.3–1 to 9) were hydrodynamically injected with pBS-WHV1.3. (A) Mouse sera were collected at the indicated time points after HI. Encapsidated WHV DNA in sera was detected by PCR. Products of the qualitative PCR were visualized by agarose gel electrophoresis. Neg.: sera from two mice (N1, 2) challenged by saline. (B) WHcAg and WHsAg expression in liver were detected by ICS. Liver samples from three mice were taken at days 0, 10, 42, and 98 after HI and stained with rabbit antibodies to WHcAg and WHsAg, respectively. A set of representative samples was shown. Magnification: 200×.</p