Objective: To explore the effect of deep dermal tissue dislocation injury on skin fibrosis in pig, in order to provide some theoretical basis for burn scar treatment. Methods: The experimental research method was applied. Six 2-month-old female Duroc pigs were taken. Fifteen operative areas on the right dorsum of pigs on which medium-thick skin grafts and deep dermal tissue slices were cut and re-implanted were included into dermal in situ reimplantation group, and fifteen operative areas on the left dorsum of pigs on which medium-thick skin grafts and deep dermal tissue slices were cut and the deep dermal tissue slice was placed under the fat layer were included into the dermal dislocation group. The hair growth in the operative areas on post-injury day (PID) 7, 14, and 21 and the cross-sectional structure on PID 14 were observed in the two groups. On PID 7, 14, and 21, the skin thickness (the distance from the epidermis to the upper edge of the fat), the dermal thickness (the distance from the lower edge of the epidermis to the upper edge of the fat, excluding the fibrotic tissue thickness between the dermis and the fat), and the fibrosis tissue thickness of the dermis-fat interface (from the lower edge of the deep dermis to the upper edge of the fat in dermal in situ reimplantation group and from the lower edge of the superficial dermis to the upper edge of the fat in dermal dislocation group) in the operative areas were measured and compared between the two groups; the fibrotic tissue thickness at the dermal cutting interface (from the lower edge of the superficial dermis to the upper edge of the deep dermis) in the operative areas in dermal in situ reimplantation group was measured and compared with the fibrotic tissue thickness at the dermal-fat interface. Sirius red staining was performed to observe and compare the type Ⅰ and Ⅲ collagen content in the dermal-fat interface in the operative areas between the 2 groups and between the dermal cutting interface and dermal-fat interface in the operative areas in dermal in situ reimplantation group. Immunohistochemical staining was performed to observe the positive expressions of proliferating cell nuclear antigen (PCNA), transforming growth factor β₁ (TGF-β₁), fibroblast growth factor 2 (FGF-2), and hepatocyte growth factor (HGF) in the operative areas in the two groups. The sample number was 6. Data were statistically analyzed with independent sample t test. Results: On PID 7, 14, and 21, the hairs in the operative areas in dermal in situ reimplantation group were denser than those in dermal dislocation group. On PID 14, the skin cross section in the operative areas in dermal dislocation group showed a "sandwich"-like structure, while the skin cross section in the operative areas in dermal in situ reimplantation group had normal structure. On PID 7, 14, and 21, the skin thickness in the operative areas in dermal dislocation group was (4 234±186), (4 688±360), and (4 548±360) μm, respectively, which was close to (4 425±156), (4 714±141), and (4 310±473) μm in dermal in situ reimplantation group (P>0.05); the dermal thickness in the operative areas in dermal dislocation group was significantly thinner than that in dermal in situ reimplantation group (with t values of -9.73, -15.85, and -15.41, respectively, P<0.01); the fibrotic tissue thickness at the dermal-fat interface in the operative areas in dermal dislocation group was significantly thicker than that in dermal in situ reimplantation group (with t values of 14.48, 20.58, and 15.67, respectively, P<0.01); there was no statistically significant difference between the fibrotic tissue thickness at the dermal-fat interface and the dermal cutting interface in the operative areas in dermal in situ reimplantation group (P>0.05). On PID 7, 14, 21, the type Ⅲ collagen content in the dermal-fat interface in the operative areas in dermal dislocation group was increased significantly compared with that in dermal in situ replantation group (with t values of 2.65, 0.61, and 7.39, respectively, P<0.05 or P<0.01), whereas there were no statistically significant differences in the type Ⅰ collagen content at the dermal-fat interface in the operative areas between the 2 groups (P>0.05) and the type Ⅰ and Ⅲ collagen content between the dermal-fat interface and the dermal cutting interface in the operative areas in dermal in situ reimplantation group (P>0.05). On PID 7, 14, and 21, PCNA, TGF-β₁, FGF-2, and HGF were positively expressed in the superficial dermis and adipose tissue in the operative areas in dermal dislocation group, while PCNA, TGF-β₁, FGF-2, and HGF were positively expressed in the superficial dermis, deep dermis, and adipose tissue in the operative areas in dermal in situ reimplantation group. Conclusions: Inadequate intrinsic thickness of dermal tissue is the key factor causing fibrosis, and the biological purpose of fibrosis is to "compensate" the intrinsic thickness of the skin. Besides, adipose tissue may also be an important component of fibrotic skin repair.
Swine ; Female ; Animals ; Dermis/pathology* ; Proliferating Cell Nuclear Antigen/metabolism* ; Fibroblast Growth Factor 2 ; Cross-Sectional Studies ; Fibrosis ; Skin Diseases/pathology* ; Collagen/metabolism*

Objective:To investigate the association between levator hiatus area, pelvic organ prolapse quantification (POP-Q) examination and prolapse symptoms.Methods:The prospective multicenter study enrolled 996 female patients between January 2017 and January 2019. All enrolled patients underwent a standard clinical interview, POP-Q examination and transperineal ultrasound examination. Volume data of pelvic floor ultrasound examinations were obtained at rest, during contraction and during maximal Valsalva maneuver. The association between levator hiatus area, POP-Q examination and prolapse symptoms was analyzed. The performance of levator hiatus area on maximal Valsalva for assessing significant POP(POP-Q stage≥2) and prolapse symptoms were also evaluated.Results:There were significant differences of levator hiatus area at rest, during contraction and during maximal Valsalva among patients with different POP-Q stages (all P<0.001). Levator hiatus area during maximal Valsalva showed the highest correlation with abdominal dragging sensation ( r=0.277, P<0.001). The area under the ROC curve (AUC) of levator hiatus area during maximal Valsalva for significant POP (POP-Q stage≥2) was significantly higher than that for prolapse symptoms (AUC: 0.77 vs 0.69, P<0.001). Conclusions:Levator hiatus area on transperineal has moderate correlation with POP-Q examination and their association is stronger than the correlation between ultrasound findings and prolapse symptoms.

Objective:To investigate the feasibility of transperineal ultrasound in quantitative assessment of posterior compartment prolapse among Chinese women.Methods:The prospective multicenter study enrolled 485 women between January 2017 and January 2019. All patients underwent a standard clinical interview, pelvic organ prolapse quantification (POP-Q) examination and transperineal ultrasound examination. Volume data of transperineal ultrasound examinations were obtained at rest and in maximal Valsalva maneuver.Results:The higher POP-Q stage of posterior compartment, the lower rectal ampulla position in maximal Valsalva maneuver (POP-Q stage=0 vs POP-Q stage=1, P<0.001; POP-Q stage=1 vs POP-Q stage≥2, P<0.001), and the greater rectal ampulla hypermobility (POP-Q stage=0 vs POP-Q stage=1, P<0.001; POP-Q stage=1 vs POP-Q stage≥ 2, P=0.007). The rectal ampulla position at rest and in maximal Valsalva maneuver and rectocele depth were correlated with prolapse symptoms ( r=-0.200, P<0.001; r=-0.252, P<0.001; r=0.086, P=0.045). The corresponding cut-off values of rectal ampulla position in maximal Valsalva in diagnosing posterior compartment prolapse (POP-Q stage ≥1) and clinical significant posterior compartment prolapse (POP-Q stage ≥2) were 7.32 mm below the symphysis pubis and 12 mm below the symphysis pubis, respectively, with the area under the ROC curve as 0.75 and 0.85, respectively. Conclusions:The ultrasonic measurements by transperineal ultrasound is significantly associated with POP-Q examination in posterior compartment, and it is demonstrated as a useful tool in quantitative assessment of the severity of posterior compartment prolapse.

Objective To summarize the clinical characteristics, follow-up and prognosis of Kawasaki disease (KD) in infants under 3 months. Method The clinical data of infants under 3 months diagnosed with KD during January 2009 to December 2013 were analyzed retrospectively. Results In a total of 1504 cases diagnosed with KD during during the study period, there were 40 (2.66%) infants under 3 months (30 males and 10 females). Except for fever, the frequencies of the other 5 main clinical manifestations were less than 50%. Laboratory tests suggested inflammatory reactions, thrombocytosis, anemia, and so on. Coronary artery disease was found in 32 cases (80%), including 24 cases of coronary artery dilatation, 8 cases of coronary artery aneurysm and 3 cases of coronary giant aneurysm. By the end of the follow-up period, there were no myocardial ischemia, myocardial infarction and death in all the patients; coronary artery diameter was normal in 37 cases (92.5%); 3 cases of giant coronary artery aneurysm still had coronary artery aneurysm and thrombosis. Conclusion The clinical manifestations of KD in little infants were atypical, the incidence of coronary artery disease is high, and color doppler echocardiography should be performed in time.

OBJECTIVETo observe the fibrosis of skin after damage to the fat dome structure in skin of pig.

METHODSTotally 4 pieces of skin grafts of intermediate thickness in the size of 5 cm × 5 cm were obtained from both sides beside the spine of back in each of the 4 female red Duroc pigs with pedicle on one side with Humby knife performed by burn specialists, who were rich in clinical experience. These skin grafts were assigned as thin dermis group (TD). Pedicled tissue grafts in the size of 5 cm × 5 cm with the thickness of 1.5 mm were obtained within the wounds resulted from former incision with the same method mentioned above, and these tissue grafts were set as fat dome group (FD). The above-mentioned two groups of skin grafts were sutured back in situ immediately after completion of the former procedures. On post surgery day (PSD) 7, 14, and 21, 5 wounds were respectively selected according to the random number table for gross observation of the surgical areas. Tissue samples were obtained from corresponding surgical area deep to the deep fascia after gross observation at above-mentioned time points. Some of the tissue samples were used for observation of distribution of collagen fibers in the regions of operation of both groups of skin grafts with HE staining, and the breadth of fibrosis was measured; some of the tissue samples were used for observation of distribution of type I or III collagen fibers in the regions of incision of both two groups of skin grafts with Sirius red staining. Data were processed with two independent sample t test.

RESULTSA little scab on the edge of wounds was observed on PSD 7; all the wounds were healed on PSD 14; a few hairs were observed growing in the surgical area on PSD 21. HE staining showed that traces of incision were observed in the superficial layer of dermis and at the junction between dermis and fat dome at each time point; profuse hyperplasia of collagen fibers with parallel and orderly arrangement were observed in the region of incision of skin grafts in groups TD and FD at each time point. The breadth of fibrosis of the region of incision of skin grafts was respectively (251 ± 31), (240 ± 3 7), and (342 ± 69) µm in group TD, (239 ± 36), (286 ± 61), and (332 ± 28) µm in group FD on PSD 7, 14, 21, without significantly statistical difference (with t values respectively 0.750, -1.971, and 0.375, P values above 0.05). Sirius red staining showed that large amount of type III collagen fibers and small amount of type I collagen fibers arranging parallelly were present in the region of incision of skin grafts in groups TD and FD at each time point.

CONCLUSIONSUnder the circumstances of relatively intact restoration of dermal tissue, no excessive fibrosis was observed after simple incisional injury of fat dome in skin of pig.

Animals ; Burns ; surgery ; Dermis ; surgery ; transplantation ; Female ; Fibrosis ; complications ; Graft Survival ; Male ; Skin ; Skin Transplantation ; methods ; Skin, Artificial ; Swine ; Wound Healing

Objective The generation of drug resistance often leads to the failure of the bladder cancer chemotherapy.P-glycoprotein (P-gp) is an ATP-dependent drug efflux pump linked to development of multidrug resistance in cancer cells.The laboratory has successfully established adriamycin-resistant human bladder cancer cell line (pumc-91/ADM) from its parental cell line (pumc-91).According to the drug resistant spectrum analysis,pumc-91/ADM cell line exhibited the characteristics of multi-drug resistance.However,the expression of P-gp in two cell lines was still unknown.In this paper,there was a comparison between pumc-91/ADM and pumc-91 about the differential expression of P-gp.Methods To determine the expression and location of P-gp in pumc-91 and pumc-91/ADM,qRT-PCR,Western blot and immunocytochemistry were applied in the experiment.qRT-PCR was implemented to research the expression of P-gp mRNA in two cell lines (pumc-91/ADM and pumc-91).Western blot was adopted to investigate the expression of P-gp protein in pumc-91 and pumc-91/ADM cell lines.Immunocytochemistry technique was used to explore the cellular location of P-gp and affirm its expression in two cell lines visually.Student's t-test was employed for statistical analysis and P ＜ 0.05 was considered statistically significant.Results qRT-PCR analysis revealed that the expression of P-gp mRNA was upregulated in drug-resistant cell line pumc-91/ADM compared to parental cell line pumc-91.To normalize for differences in the amount of total RNA,GAPDH was selected as an endogenous RNA control.Compared with pumc-91,the expression of P-gp mRNA was upregulated 7.74 fold in pumc-91/ADM (t =11.97,P ＜ 0.05).Consistent with the qRT-PCR result,Western blot confirmed the protein of P-gp expressed differentially in two cell lines.The expression of P-gp protein was significantly increased in pumc-91/ADM compared to pumc-91.According to the results,the differences between pumc-91 and pumc-91/ADM had statistical significance (t =4.35,P＜0.05).Immunocytochemical analysis results demonstrated that P-gp was not only located in cell membrane but also in cytoplasm of the two cell lines.The expression of P-gp in pumc-91/ADM increased distinctly.The difference was statistically significant (t =11.41,P ＜ 0.05).Conclusion Compared with pumc-91,the expression of P-gp in pumc-91/ADM was significantly upregulated.

Objective To investigate the expression of human papillomavirus E2 mRNA in cervical cytologi-cal specimens as well as the role of that in cervical carcinogenesis and its clinical significance in screening and evalat-ing prognosis of cervical lesions.Methods The expression of E2 mRNA in 582 cases of cervical cytological speci-mens with high risk human papillomavirus infection and cytological diagnosis of NILM,ASCUS and LSIL,was detected by RT -PCR.Thereafter,all cases were divided into the four groups by colposcopy and histopathological confirmation, including 414 cases of cervicitis,95 cases of CINⅠ,51 cases of CINⅡ,20 cases of CINⅢ and 2 cases of invasive cervical carcinoma as well.Results The expression of HPV -E2 mRNA decreased dramatically corresponding with pathological upgrading from groups of cervicitis to invasive cervical carcinoma (χ2 =132.72,P <0.05).The sensitiv-ity,specificity,positive predictive value and negative predictive value of HPV -E2 mRNA for screening potential cervical lesions in group of NILM,ASCUS and LSIL were 77.2%,96.8%,75.6%,96.0% and 81.0%,91.7%, 79.1%,92.6% as well as 95.9%,93.4%,94.7%,95.0% respectively.Conclusion Deletion of HPV -E2 induced by genetic disruption played an important role in the early stage of malignant transformation of cervical epithe-lial cells.Therefore,detection of the levels of HPV -E2 mRNA expression might be clinically valuable for the screen-ing and evaluating of prognosis in cervical lesions.

Objective To investigate the role and signalling of microRNA (miR)-182 on regulating high glucose-induced cardiomyocyte hypertrophy.Methods The candidates of miR which might potentially be involved on targeting Rac1 were predicted by applying bioinformatics analysis.The expression of all related candidates miRs was verified by real-time reverse transcription-PCR(RT-PCR)in cardiac tissues of db/db mice and db/m mice.Then the relationship between candidates miR and Rac1 was investigated with Pearson relevant analysis.Neonatal mice cardiomyocytes were cultured and divided into 2 groups:normal glucose group and high glucose group.The level of selected miR and Rac1 in two groups was detected by RT-PCR.Neonatal mice cardiomyocytes were then randomly divided into 4 groups:normal glucose group,selected microRNA mimics control group,high glucose group,high glucose plus selected miR mimics control group.The morphology of cardiomyocyte in each group was detected under light microscope.Furthermore,Rac1,β-MHC and α-SMA expressions were detected in cultured cardiomyocyte treated by high glucose for 48 h after transfecting selected miR mimics by RT-PCR and Western blot.Results A total of 6 miR candidates potentially targeting Rac1 were screened by bioinformatics,which were miR-182,miR-142-3p,miR-140,miR-101a,miR-429 and miR-200b.Among these candidates,miR-182 and miR-142-3p expression was significantly downregulated in cardiac tissues of db/db mice compared with db/m controls (P ＜0.05).MiR-182 was negatively correlated with Rac1 by person analysis (r =-0.891 02).Downregulation of miR-182 and upregulation of Rac1,β-MHC,α-SMA were found in high glucose-induced cardiomyocyte.After transfection of miR-182 mimics,hypertrophic changes were significantly reduced and Rac1 as well β-MHC expression was significantly downregulated in cardiomyocyte incubated with high glucose.Conclusion MiR-182 might be involved in the regulation of high glucose-induced myocardial hypertrophy process via targeting Rac1.

Objective To describe early clinical experience with the new amplatzer ductal occluder Ⅱ (ADO Ⅱ) for transcatheter patent ductus arteriosus(PDA) in children.Methods Twelve children were diagnosed as PDA from Jan.2013 to Apr.2014 in Guangzhou Children and Women's Hospital.All the children who were treated with the ADO Ⅱ had the indication of a successful interventional therapy successfully.The size of device was chosen according to aortographic and transthoracic echocardiography(TTE) results and pulmonary pressure.The device was delivered in a consequent or reverse way depending on the type of PDA,the minimal diameter of PDA and the size of duct ampulla.The device was delivered in a reverse way in ten patients,and two in a consequent way before detaching the device.Another aortogram was performed in order to check the position and form of the device,the velocities of blood flow in left pulmonary artery and the descending aorta though TTE and whether there was a residual shunt.All patients were examined by TTE in 24 hours after surgery and discharged without any complications 2 days later.The patients were programmed for the cardiologic consult including an TTE and electrocardiogram in 1,3,6 and 12 months after discharge.Results Twelve patients(7 male,5 female) with a median age of(1.59 ± 1.10) years(range 0.53-4.47 years),a median weight of (9.52 ± 3.41) kg(range 5.5-18.3 kg),a median pulmonary blood flow/systemic blood flow (Qp/ Qs) of 1.64 ± 0.45 (range 1.33-2.85),a median pulmonary artery systolic pressure (32.50 ± 10.05) mmHg (range 23-58 mmHg,1 mmHg =0.133 kPa),and the minimum (2.40 ±0.68) mm (1.6-3.8 mm),underwent transcatheter ductal closure with the ADO Ⅱ.Device sizes used were 3 mm ×4 mm(n =7),3 mm ×6 mm(n =3),6 mm ×6 mm (n =2),respectively and delivered with 4 or 5 F delivery catheters.The median fluoroscopy time was (6.39 ± 4.16) min(range 3.2-18.2 min).Complete ductal occlusion was achieved by the end of the procedure in 10 patients.The TTE showed good position of the occlusion and the velocities of blood flow in left pulmonary artery and the descending aorta were in a normal range.There was a trivial residual shunt after the surgery of 2 patients.No residual shunt was found after 24 hours in all 12 patients.In 1 case,the patient had a descending aortic obstruction with pressure gradient of 11 mmHg.Three months after surgery,the pressure descended to 10 mmHg by TTE.Complete ductal occlusion without aortic arch or left pulmonary artery stenosis had been identified in other 11 remaining patients on TTE follow-up of 6 months of 3 patients and 12 months of 6 patients.Conclusions The ADO Ⅱ achieves excellent ductal closure rates through low profile delivery systems in small infants and children with moderate and small PDA or morphologically varied PDAs.It is simple in use with few complications.Occlusion design allows closure with arterial or venous approach and delivery with 4 or 5 F delivery catheters.The children who used arterial approach,transthoracic echocardiography TTE is recommended to replace aortic angiography,so as to avoid puncturing the aorta and reduce vascular injury.

Objective To discuss the meaning of continuous intracranial pressure (ICP) monitoring in patients with severe traumatic craniocerebral injury. Methods One hundred and twenty four patients with severe craniocerebral injury treated from August 2004 to February 2011 in our hospital, were enrolled and divided randomly into ICP monitoring group (n = 62) and routine treatment group (n = 62). The patients of ICP monitoring group had adjusted treatment plan according to the changes of ICP at any time, whereas the patients in routine treatment group underwent routine neurosurgical treatment according to the doctors' experience. Results There were 7 cases of acute kidney function failure,and 11 cases of electrolyte disturbances in the ICP group.There were 15 cases of acute kidney function failure, and 25 cases of electrolyte disturbances in the routine group. The complication rate in the ICP group was lower than that in the routine group (x2 =3. 54 and 7.67 for acute kidney function failure and electrolyte disturbances respectively, Ps <0. 01). The days of mannite using were (6±2)dand (15 ±3)d, respectively; the dosage of mannite using were (749 ± 125) g and (1545 ±250) g,respectively. The good recovery and slight disability were 28 and 16 cases in the ICP group, and 13 and 9 cases in the routine group,respectively. The severe disability,vegetative state and death were 9,4 and 8 cases in the ICP group,and 17,7 and 13 cases in the routine group. The days and dosage of mannite using in the ICP group were much less than those in the routine group (t = 8. 32 and 7.41, Ps < 0. 01). The proportion of good recovery and slight disability in the ICP group were higher than those in the routine group(x2 =5. 07 and 3. 55,Ps <0.01). However, the proportion of severe disability, vegetative state and death in the ICP group were lower than those in the routine group (x2 =0.84,0.89 and 1.43, Ps < 0. 01) . Conclusion Continuous ICP monitoring in severe craniocerebral injury shows benefits in directing treatment plan adjustment, reducing complications and improving the prognosis.