Objective To evaluate the effects of shRNA-TGF-β1 plasmid on Smads signal transduction of rat renal allograft.Methods A Sprague-Dawley to Wistar rat orthotopic transplant kidney-sclerosis accelerated model was constructed and transfected with short hairpin RNA-TGF-β1 based on the hydromechanics.The recipients were divided into three groups:group T(plasmid group)injected with shRNA-TGF-β1;group H(vacant plasmid group)injected with vacant plasmid;group Y(simply transplantation group)injected with no plasmid.In group J(sham-operated group)only right kidney was removed with no transplantation as control group.Transplanted kidneys and blood samples were collected at the first,second and third month after transplantation.The blood urea nitrogen(BUN)and serum Cr were tested by enzyme-linked immunoadsordent assay.The gene transcriptional level of TGF-β1 and Smad3/7 was detected by RT-PCR,and the protein variations of TGF-β1 and phosphorylated Smad3/7 were examined by Western blotting.Results At each test time point,the BUN and serum Cr were significantly higher in the plasmid group than in the sham-operated group(P<0.05 or P<0.01),but obviously lower than in the vacant plasmid group and simply transplantation group(P<0.05 or P<0.01).The expression of TGF-β1 as well as phosphorylated Smad3 was significantly higher in the plasmid group than in the sham-operated group(P<0.05 or P<0.01),but obviously lower than in the vacant plasmid group and simply transplantation group(P<0.05 or P<0.01).However,the expression of phosphorylated Smad7 was significantly lower in the plasmid group than in the sham-operated group(P<0.05 or P<0.01),but obviously higher than in the vacant plasmid group and simply transplantation group(P<0.05 or P<0.01).Conclusion Short hairpin RNA-TGF-β1 plasmid could significantly improve the renal function of rat renal allografts probably by downregulating phosphorylated Smad3 and upregulating phosphorylated Smad7,leading to the inhibition of TGF-beta 1 promoting fibrosis role and delay of the allograft fibrosis.

Objective To explore the mechanism of Niaochangshu capsule in the treatment of postmenopausal overactive bladder, through observing its influence on bladder weight and the expression of eNOS and AQP1 of ovariectomized female rats. Methods Female SD rats were divided into blank group, model group, Nilestriol group and Niaochangshu group. Rats were removed ovaries except the blank group. The treatment groups were given corresponding drugs, blank group and model group were given normal saline by gavage. After 4 weeks, the bladders' weight and thickness were detected, the expressions of eNOS and AQP1 in serum and bladder tissue were determined by ELISA, and NO by spectrophotometry. Results Ovariectomy resulted in decreased bladder weight, bladder mucosal and muscular atrophy, and opposite changes showed after given Niaochangshu. The expressions of eNOS and NO in bladder and serum were decreased significantly after ovariectomy, while increased by given Niaochangshu capsule or nylestriol (P<0.05), and there was significant difference between Niaochangshu group and Nilestriol group (P<0.05). The expression of AQP1 was decreased in the model group, and increased after given nylestriol or Niaochangshu capsule. While the expression of AQP1 in bladder had no significant difference among the four groups. Conclusion Niaochangshu capsule can reverse bladder mucosal and muscular atrophy caused by estrogen deficiency, and increase the content of eNOS in serum and bladder, thus play the role in the treatment of postmenopausal overactive bladder.

This study prospectively examined the intranasal distribution of nasal spray after nasal septal correction and decongestant administration. A cohort of 20 patients was assessed for the distribution of nasal spray before and after nasal septum surgery. Sprays were dyed and administered one puff per nostril when patients hold their head up in an upright position. Before and after decongestant administration, the intranasal distribution was semi-quantitatively determined by nasal endoscopy. The results showed that the dyed drug was preferentially sprayed onto the nasal vestibule, the head of the inferior turbinate, the anterior part of septum and nasal floor. As far as the anterior-inferior segment of the nasal cavity was concerned, the distribution was found to be influenced neither by the decongestant nor by the surgery (P>0.05). However, both the decongestant and surgery expanded the distribution to the anatomical structures in the superior and posterior nasal cavity such as olfactory fissure, middle turbinate head and middle nasal meatus. No distribution was observed in the sphenoethmoidal recess, posterior septum, tail of inferior turbinate and nasopharynx. It was concluded that nasal septum surgery and decongestant administration significantly improves nasal spray distribution in the nasal cavity.

Objective: To analyze the characteristics and treatment of diabetic patients with superficial partial-thickness burn on feet. Methods: Eighty-three patients with superficial partial-thickness burn on 119 feet were hospitalized in our unit from January 2011 to December 2017. The medical records of the patients with 46 men and 37 women, aged 60±11 were retrospectively analyzed. The patients were divided into diabetes group and non-diabetes group according to whether they had diabetes or not, with 41 patients (60 burn feet) in diabetes group and 42 patients (59 burn feet) in non-diabetes group. Patients in diabetes group and non-diabetes group were given systemic treatment and wound dressing change. Thirty-seven diabetic patients whose wounds deepened to deep partial-thickness were divided into eschar shaving group and non-eschar shaving group according to patients′ willingness and the treatment, with 14 patients in eschar shaving group and 23 patients in non-eschar shaving group. Patients in eschar shaving group were given eschar shaving operation at early stage, and patients in non-eschar shaving group were given wound dressing change. The length of hospital stay, hospitalization treatment expenses, pulse of arteria dorsal pedis and posterior tibial artery immediately after admission, deepening of wounds on feet during hospital stay, and rates of wound healing on feet of patients in diabetes group and non-diabetes group were observed and calculated. Pulses of arteria dorsal pedis and posterior tibial artery immediately after admission, deepening of wounds on feet during hospital stay, positive rates of bacteria and fungus in wounds on feet, and rates of wound healing on feet of patients in eschar shaving group and non-eschar shaving group were observed and calculated. Data were processed with chi-square test, t test, Fisher′s exact propability method, and Mann-Whitney U test. Results: The length of hospital stay of patients in diabetes group was (29±20) d, which was significantly longer than that of patients in non-diabetes group [(19±13) d, t=2.730, P<0.01]. The hospitalization treatment expense of patients in diabetes group was (46 988±41 322) yuan, which was significantly more than that of patients in non-diabetes group [(29 106±24 813) yuan, t=2.396, P<0.05]. The pulses of arteria dorsal pedis and posterior tibial artery of patients in diabetes group were significantly weaker than those of patients in non-diabetes group (Z=3.278, 2.194, P<0.05 or P<0.01). The percentages of wounds on feet of patients in diabetes group deepening to deep partial-thickness burn, full-thickness skin defect with bone and tendon exposure were respectively 88.3% (53/60) and 23.3% (14/60), which were significantly higher than those of patients in non-diabetes group [47.5% (28/59) and 1.7% (1/59), χ²=22.867, 12.644, P<0.01]. Rate of wound healing on feet of patients in diabetes group was 78.3% (47/60), which was significantly lower than 100.0% (59/59) of patients in non-diabetes group ( χ²=14.351, P<0.01). There were respectively 21 and 32 feet in patients of eschar shaving group and non-eschar shaving group. There were no significantly statistical differences in pulses of arteria dorsal pedis and posterior tibial artery of patients between eschar shaving group and non-eschar shaving group (Z=0, 0.453, P>0.05). The percentage of wounds on feet of patients in non-eschar shaving group deepening to full-thickness skin defect with bone and tendon exposure was 43.8% (14/32), which was significantly higher than 0 of patients in eschar shaving group ( χ²=12.486, P<0.01). Positive rates of bacteria and fungus in wounds on feet of patients in eschar shaving group was significantly lower than that of patients in non-eschar shaving group (χ²=4.386, P<0.05 ). Rate of wound healing on feet of patients in non-eschar shaving group was 59.4% (19/32), which was significantly lower than that of patients in eschar shaving group [100.0% (21/21), P<0.01]. Conclusions Diabetes patients with superficial partial-thickness burn wounds on feet has long length of hospital stay, high hospitalization treatment expenses. Wounds of the patients are easy to deepen, with low wound healing rate. Eschar shaving at early stage when the wounds deepened to deep partial-thickness burn is a good way to increase wound healing rate and prevent further deepening of wounds.

This study prospectively examined the intranasal distribution of nasal spray after nasal septal correction and decongestant administration.A cohort of 20 patients was assessed for the distribution of nasal spray before and after nasal septum surgery.Sprays were dyed and administered one puff per nostril when patients hold their head up in an upright position.Before and after decongestant administration,the intranasal distribution was semi-quantitatively determined by nasal endoscopy.The results showed that the dyed drug was preferentially sprayed onto the nasal vestibule,the head of the inferior turbinate,the anterior part of septum and nasal floor.As far as the anterior-inferior segment of the nasal cavity was concerned,the distribution was found to be influenced neither by the decongestant nor by the surgery (P＞0.05).However,both the decongestant and surgery expanded the distribution to the anatomical structures in the superior and posterior nasal cavity such as olfactory fissure,middle turbinate head and middle nasal meatus.No distribution was observed in the sphenoethmoidal recess,posterior septum,tail of inferior turbinate and nasopharynx.It was concluded that nasal septum surgery and decongestant administration significantly improves nasal spray distribution in the nasal cavity.

Objective To summarize the experience with cadaveric renal transplantation for improving the long-term survival rate of the recipients.Methods The clinical data of 1210 cases(773 men and 437 women;age range,6-75 years) of cadaveric kidney transplantation from 1986 to 2003 were analyzed retrospectively,including the resection of the donor's kidneys,surgical techniques,use of immunosuppressants,and complications.The 1210 patients underwent renal transplantation for most of them(1047 cases) suffered from chronic glomerulonephritis.Lymphocytotoxicity test was performed in 1210 cases with all

Here, we presented a method to bacterially express the major structural protein L1 of Human Papillomavirus type 18 (HPV18) as soluble form. We found that the purified L1 could self-assemble to virus-like particles (VLPs). Further, we investigated the immunogenicity and the induced level of neutralizing antibody using these VLPs. First, the genome of HPV18 was cloned from a patient in Xiamen. It was used as template for PCR amplification of HPV18 L1 gene. The resultant DNA fragment was inserted into expression vector pTrxFus and expressed in Escherichia coli GI724. Second, L1 protein was purified by ammonium sulfate precipitation, ion-exchange chromatography and hydrophobic interaction chromatography; and the purified L1 was subjected to self-assembly to form VLPs with the removal of premixed reductant DTT. Finally, the size and morphology of these VLPs was investigated by Dynamic Light Scattering and Transmission Electronic Microscopy as 29.34 nm in hydrated radius and globular particles similar with native HPV18. The half effective dosage (ED50) and maximum level of neutralizing antibody elicitation were measured by vaccinations on mice, rabbit and goat using pseudovirus neutralization cell model. The results showed that the ED50 of HPV18 VLPs is 0.006 microg in mice, and the maximum titer of neutralizing antibody elicited in rabbit and goat is up to 10(7). As a conclusion, we can provide HPV18 VLPs with highly immunogenicity from prokaryote expression system, which may pave a new way for research and development of prophylactic vaccine for HPV18.
Animals ; Capsid Proteins ; biosynthesis ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Goats ; Human papillomavirus 18 ; immunology ; isolation & purification ; Mice ; Oncogene Proteins, Viral ; biosynthesis ; genetics ; immunology ; Rabbits ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Virion ; genetics ; immunology

ObjectiveBased on ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry （UPLC-Q-TOF/MS）， the changes of endogenous markers in rat plasma at the different stage， namely modeling and administration of Shenling Baizhusan （SLBZS）， and the mechanism of SLBZS in the treatment of ulcerative colitis （UC） were studied. MethodIn the modeling stage， rats were randomly divided into normal group， spleen deficiency with dampness retention-UC （SDDR-UC） and pure-UC （P-UC） model group. In the administration stage， SLBZS was given to the above two different model groups. After modeling and administration， rat plasma was collected and determined by UPLC-Q-TOF/MS. The mobile phase was 0.1% formic acid aqueous solution （A）-acetonitrile （B） for gradient elution （in positive ion mode：0-2 min， 99%A； 2-9 min， 99%-73%A； 9-10 min， 73%-44%A； 10-13 min， 44%-38%A； 13-19 min， 38%-28%A； 19-21 min， 28%-2%A； 21-23 min， 2%A； 23-25 min， 2%-10%A； 25-27 min， 10%-99%A； in negative ion mode：0-2 min， 85%A； 2-3 min， 85%-65%A； 3-5.5 min， 65%-44%A； 5.5-8 min， 44%-25%A； 8-10 min， 25%-2%A； 10-16 min， 2%-85%A）. The electrospray ionization （ESI） temperature was 120 ℃ under the positive and negative ion modes， and the acquisition range was 50-1 000. Partial least squares-discriminant analysis （PLS-DA） was used to analyze the changes of endogenous metabolites in the above two different model rats from the different stage. MetaboAnalyst 5.0 was used to analyze the metabolic pathways of these identified metabolites. ResultSixteen potential biomarkers were screened and identified in the modeling stage， among which 11 potential biomarkers were common in the two model rats， which mainly affected the primary bile acid biosynthesis pathway. Twenty-three potential biomarkers were screened and identified during the administration stage， among which 3 potential biomarkers were shared by the two model rats， and SDDR-UC and P-UC model rats had 11 and 9 potential biomarkers， respectively. It mainly affected 6 pathways such as purine metabolism， pentose phosphate pathway， pyrimidine metabolism， retinol metabolism， primary bile acid biosynthesis and steroid hormone synthesis. ConclusionThe primary bile acid biosynthesis pathway appears in the different stage of modeling and administration of UC， showing a dynamic change process. The therapeutic effect of SLBZS on SDDR-UC rats may be related to inhibiting the expression of nuclear transcription factor -κB （NF-κB） signaling pathway， activating farnesoid X receptor （FXR） and enhancing the expression of cytochrome P450.