Objective To explore the correlation between cytomegalovirus pneumonia (CMV-IP) and viral load in renal transplant recipients and to find out the threshold value of viral load for predicting CMV-IP. Methods The blood samples of 56 renal transplant recipients were taken weekly for the first 2 months and every 2 weeks for 2-6 months after transplantation.Real-time PCR were used to quantify the plasma CMV DNA.The mean viral loads of CMV-IP group and non-CMV-IP group in each time were compared using Wilcoxon test.Different cut-off values were tested to find the suitable values to predict the CMV-IP. Results Of the 56 recipients,8 (14.3%) developed CMV-IP.The viral loads were near zero in the first 4 weeks in both groups;from week 5 the viral load of CMV-IP group increased gradually and reached the climax at week 8 and then declined,but the load of non-CMV-IP group fluctuated at a low level.During weeks 5-11,the viral loads of CMV-IP group were higher than those of non-VMV-IP group.At 5,7 and 9 weeks,the differences of the viral loads between the 2 groups were significant (P

OBJECTIVE: To observe the effect of different frequencies (2 Hz, 100 Hz) of electroacupuncture (EA) on limb locomotion and the expression of inflammatory factors IL-1β, IL-6, TNF-α in sciatic nerve, and nuclear factor kappa B (NF-κB) in lumber(L)4-L5of spinal cord in rats with sciatic nerve injury (SNI), so as to reveal its mechanisms underlying improvement of SNI. METHODS: A total of 48 SD rats (half male and half female) were equally divided into blank control, model, low frequency (2 Hz) EA and high frequency (100 Hz) EA groups. The SNI model was established by clamping the spinal nerve. EA intervention (2 Hz, 100 Hz, 1 mA), starting on the 8th day after modeling, was applied to "Huantiao" (GB30) on the injured side for 15 min, once daily for 14 consecutive days. The sciatic function index (SFI) was calculated to assess the injured hindlimb recovery with reference to BAIN's and colleagues' methods. Histopathological changes of the sciatic nerve were displayed by H.E. staining. The expressions of IL-1β, IL-6 and TNF-α in the sciatic nerve tissue were detected by immunohistochemistry, and the expression of NF-κB in the spinal cord was detected by using Western blot. RESULTS: After modeling, the SFI level on day 8 was significantly decreased in the model group (P<0.01), and no significant differences were found among the model, low frequency EA and high frequency EA groups before the EA intervention (P>0.05). Following the treatment (at the 22nd day), the SFI values of both low frequency EA and high frequency EA groups were significantly increased (P<0.01), suggesting an improvement of the limb motor function, and the SFI of the low frequency EA group was notably higher than that of the high frequency EA group (P<0.01). In comparison with the blank control group, the expression levels of IL-1β, IL-6, TNF-α in the sciatic nerve and NF-κB protein in the spinal cord were significantly up-regulated (P<0.05). Following EA intervention, the increased expression levels of IL-1β, IL-6, TNF-α and NF-κB proteins were significantly down-regulated in both low frequency EA and high frequency EA groups (P<0.05), and the therapeutic effect of low frequency EA was markedly superior to that of high frequency EA in down-regulating the expression levels of IL-1β, IL-6, TNF-α and NF-κB protein (P<0.05). H.E. staining showed increase of Schwann cells in number, cellular swelling, and disintegration of the axons and myelin sheath, and appearance of vacuolar degeneration in the model group, which was relatively milder in both low frequency EA and high frequency EA groups, particularly in the low frequency EA group. CONCLUSION: EA of GB30 at 2 Hz and 100 Hz can promote the recovery of hindlimb motor function in SNI rats, which is probably related to its function in inhibiting the inflammatory response, and facilitating the repair of the damaged sciatic nerve. 2 Hz EA is better than 100 Hz EA in the therapeutic effect.

OBJECTIVETo observe the effect of Zhibai Dihuang Decoction (ZDD) on mRNA and protein expressions of transient receptor potential family vanilloid subtype 1 (TRPV1) and transient receptor potential family vanilloid subtype 5 (TRPV5) in Ureaplasma urealyticum (UU)-infected rat semens and spermatogenic cells, and to explore the pathomechanism of UU-infected infertility and the intervention of ZDD.

METHODSTotally 45 were randomly selected from 60 4-5 months old SD rats. UU testicular infected animal models were set up after bladder inoculation of UU suspension. The remaining 15 rats were simultaneously injected with normal saline as a normal control group. After a successful modeling, UU infected model rats were randomly divided into the model group, the azithromycin group, and the ZDD group, 15 in each group. Rats in the ZDD group were administered with ZDD at the daily dose of 1 g/kg by gastrogavage. Rats in the azithromycin group were administered with azithromycin suspension at the daily dose of 0. 105 mg/kg by gastrogavage. Equal volume of normal saline was administered to rats in the normal control group and the model group. All medication was performed once daily for 21 successive days. Testes and epididymis were extracted after rats were killed and UU positive rates were compared among all groups. Sperm cells were separated using a mechanical separation technique. Sperm motility parameters were detected using color sperm motion detection system. mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells were determined by real-time quantitative PCR and Western blot.

RESULTSThe UU positive rate was obviously higher in the model group than in the normal control group [(86.7% (13/15 cases) vs. 0] P < 0.05). It was lower in the ZDD group [33.3% (5/15 cases)] and the azithromycin group [26.7% (4/15 cases)] than in the model group (P < 0.05). Compared with the normal control group, class A and B sperms were reduced, the linear velocity and the average velocity were significantly lowered, mRNA and protein expressions of TRPV1 and TRPV5 in spermated genic cells significantly decreased in the model group (P < 0.05). Compared with the model group, class A and B sperms were increased, linear and curve velocities and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group and the azithromycin group (P < 0.05, P < 0.01). Compared with azithromycin group, class A and B sperms were increased, the linear velocity and the average velocity were significantly elevated, mRNA and protein expressions of TRPV1 and TRPV5 significantly increased in the ZDD group (P < 0.05, P < 0.01).

CONCLUSIONZDD could fight against UU infection and elevate semen quality, which might be associated with up-regulating mRNA and protein expressions of TRPV1 and TRPV5 in spermatogenic cells.

Animals ; Calcium Channels ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Infertility ; Male ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Semen Analysis ; Sperm Motility ; Spermatozoa ; TRPV Cation Channels ; metabolism ; Testis ; Ureaplasma Infections ; Ureaplasma urealyticum

Objective To study the effects of curcumin mediated photodynamic therapy (PDT)on the growth and proliferation in human cervical cancer cell line H8 in vitro and in vivo,and to investigate its antitumor mechanisms.Methods The effects of curcumin mediated PDT on proliferation of human cervical cancer H8 cell by MTT assay was used to screen the optimal parameter.Changes in cell morphology were observed by May-Gr ünwald-Farbstoff Giemsa staining.The apoptosis rate was estimated by flow cytometry.The effect of PDT by curcumin on the expressions of Bcl-2,P53 and survivin in H8 cells was detected by fluorescence real-time reverse transcription-polymerase chain reaction (RT-PCR).Forty BALB/C nude mice underwent subcutaneous injection of H8 cell line so as to establish animal models,and then were randomly divided into four equal groups:control group,irradiation alone group,curcumin alone group,curcumin PDT group.HE staining and pathological examination were performed.Immunohistochemical study was conducted to detect the protein expression of the apoptosis inhibiting genes of Bcl-2.Results The proliferation inhibition of H8 cells was obvious after PDT when curcumin 5μmol/L with irradiation 100 J/cm2,and with dose dependent manner.Typical morphologic features of apoptosis appeared characterizedly by marked chromatin condensation,nuclear pyknosis and fragmentation,and the appearance of apoptotic bodies.The total apoptosis rate was higher in PDT group [(47.21 ± 4.11)％]than in control group(1.71 ±0.16) ％ (P＜0.01).The mRNA expression of Bcl-2,P53 and survivin in H8 cells were suppressed significantly.HE staining showed remarkable subcutaneous necrosis in the PDT group.Immunohistochemistry showed remarkable down-regulation of protein expression of Bcl-2(P＜0.01).Conclusions Curcumin-mediated photodynamic therapy has a significant killing effect on H8 cells in vivo and in vitro.Its antitumor effect might be related to induction of Tumor cell apoptosis and suppression of Bcl-2 mRNA and protein expression.

Objective To investigate the tendency of distribution and drug-resistance of the causative organisms of urinary tract infections(UTIs)in Wuhan,and provide reliable evidence for clinical treatment.Methods Analyzed the 5 378 stains of pathogen isolated from the urine of patients in hospital.The bacteria isolates were identified with BD Phoenix-100 while can-dida isolates were identified by color plate.Results A total of 5 378 stains of pathogen had been isolated.There were 2 945 stains (54.8%)of Gram-negative bacteria,1 657 stains (30.8%)of Gram-positive bacteria,776 stains (14.4%)of fungus. The rates of Escherichiacoli resistant to penicillin were highest (>83%),and there were no carbapenem-resistant strains. There were vancomycin and linezolid-resistant Enterococcispp strains,the lowest dection rates of which were 0.3%.The de-tection rate of MRCNS was over 83%.Conclusion Escherichiacoli was the most common pathogens of urinary tract infec-tion,and theβ-lactamase inhibitor complex can be used as empirical treatment of E.coli infections.Thedetection rate of MRCNS increased,which shoud be kept a watchful eye on.

OBJECTIVETo study the effect of Zhibai Dihuang Pill (ZBDHP) on urokinase-type plasminogen activator (uPA) and sperm quality in ureaplasma urealyticum (Uu) infection infertile patients.

METHODSRecruited were 80 infertility patients with Uu infection at Andriatrics Clinics and Department of Reproduction, including 130 cases of positive Uu semen and 50 cases of negative Uu semen. Patients with positive Uu semen were randomly assigned to the observation group (72 cases) and the control group (58 cases) according to the visit sequence. All patients took antibiotics for 2 weeks. Patients in the observation group additionally took ZBDHP, 6 g each time, twice daily. Those in the control group additionally took Vit E (100 mg each time, twice per day) and ATP (40 mg each time, twice per day). The therapeutic course for all was 90 days. Semen parameters and uPA contents of the sperm membrane were detected and comparatively analyzed.

RESULTSThe sperm membrane uPA content, the sperm motility, the sperm viability, and the percentage of normal morphology sperm in Uu positive infected patients were lower than those in Uu negative infected patients with statistical difference (P < 0.05), but with no significant difference in the sperm density between the two groups (P > 0.05). There was no statistical difference in pre-treatment sperm membrane uPA contents and sperm parameters between the two groups (P > 0.05). Compared with before treatment in the same group, the sperm membrane uPA content, the sperm motility, the sperm viability, and the percentage of normal morphology sperm obviously increased in the two groups with statistical difference (P < 0.05). After treatment, the sperm membrane uPA content increased more obviously in the observation group, with statistical difference when compared with the control group (P < 0.05).

CONCLUSIONSInfection with Uu leads to decreased uPA content of sperm membrance and the sperm motility. ZBDHP could effectively treat Uu infected infertility possibly through fighting against Uu damaged sperm membrane and make the sperm membrane uPA content return to normal, and elevate the fertilizability of sperms.

Anti-Bacterial Agents ; administration & dosage ; pharmacology ; therapeutic use ; Communicable Diseases ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Humans ; Infertility ; Infertility, Male ; Male ; Semen ; Semen Analysis ; Sperm Count ; Sperm Motility ; Spermatozoa ; Ureaplasma Infections ; drug therapy ; Ureaplasma urealyticum ; drug effects ; Urokinase-Type Plasminogen Activator ; metabolism

The total RNA was extracted from porcine ovary. Porcine Follistatin cDNA was cloned by RT-PCR. Complete porcine follistatin cDNA coding sequences are presented including 1038 bp of open reading frame. The purified porcine follistatin cDNA was inserted into pGEX-4T-3 vector to construct the prokaryotic fusion protein expression vector. The recombinant expression plasmid was transformed into BL21 (DE3) and expression was induced by IPTG. Protein products were detected by SDS-PAGE and confirmed by Western blotting analysis, which showed that the yield of the Follistatin cDNA was a 63kD protein expression vector. Follistatin protein was expressed in the form of glutathione-S-transferase (GST) fusion protein in E. coli.
Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; Escherichia coli ; genetics ; Follistatin ; chemistry ; genetics ; Molecular Sequence Data ; Phylogeny ; Recombinant Fusion Proteins ; biosynthesis ; Swine

Angiotensin I-converting enzyme (ACE) inhibitory peptides have been shown to have antihypertensive effects and have been utilized for physiologically functional foods and pharmaceuticals. The ACE inhibitory ability of a hydrolysate is determined by its peptide composition. However, the peptide composition of a hydrolysate depends on proteolytic enzyme and the hydrolysis conditions. In this study, the effect of process conditions on the ACE inhibitory activity of rice dregs hydrolyzed with a trypsin was investigated systematically using response surface methodology. It was shown that the ACE inhibitory activity of rice dregs hydrolysates could be controlled by regulation of five process conditions. Hydrolysis conditions for optimal ACE inhibition were defined using the response surface model of fractional factorial design (FFD), steepest ascent design, and central composite design (CCD).
Angiotensin-Converting Enzyme Inhibitors ; analysis ; chemistry ; Combinatorial Chemistry Techniques ; methods ; Drug Evaluation, Preclinical ; Enzyme Activation ; Hydrolysis ; Oryza ; chemistry ; Peptidyl-Dipeptidase A ; chemistry ; Plant Extracts ; chemistry ; Plant Proteins ; chemistry ; Protein Hydrolysates ; chemistry

OBJECTIVETo investigate the protective effect of tea polyphenols against testis injury induced by unilateral testicular torsion/detorsion.

METHODSTwenty-four healthy male Wistar rats were equally randomized into Group I, sham operation, and Groups II and III, subjected to left lateral 720 degrees testicular torsion, followed by detorsion at 6 hours. Intraperitoneal injection of isotonic saline and polyphenols was initiated 30 minutes prior to detorsion and maintained at a low dose for 3 days postoperatively. All the rats were fed under the same condition and sacrificed 5 days later, the left torsional testes harvested for the detection of the levels of malondialdehyde (MDA) and superoxide dismutase (SOD) and the apoptosis of spermatogenic cells by TUNEL.

RESULTSSignificant differences were observed among Groups I, I and III in the levels of SOD in the left torsional testes ([285.00 +/- 22.51], [242.00 +/- 17.62] and [261.00 +/- 10.01] nU/mg, P < 0.05), as well as in the levels of MDA ([1.81 +/- 0.20], [4.34 +/- 0.34] and [2.94 +/- 0.38] nU/mg, P < 0.05). And the apoptosis indexes of spermatogenic cells were 6.64 +/- 1.82, 55.23 +/- 6.46 and 31.84 +/- 5.56 in the three groups, significantly reduced in Group III as compared with Group II (P < 0.05).

CONCLUSIONTea polyphenols has a protective effect against testicular torsion-induced ischemic reperfusion injury in rats.

Animals ; Apoptosis ; Flavonoids ; pharmacology ; Male ; Malondialdehyde ; metabolism ; Phenols ; pharmacology ; Polyphenols ; Rats ; Rats, Wistar ; Spermatic Cord Torsion ; metabolism ; pathology ; Superoxide Dismutase ; metabolism ; Tea ; chemistry ; Testis ; drug effects ; metabolism ; pathology

BACKGROUND:Anti-infective ability determine the success or failure of skin grafting. It is one of the commonly used methods to enhance the anti-infective ability of implants by compounding antibacterial materials with scaffolds. OBJECTIVE:To investigate the effect of porous collagen/silk fibroin scaffolds carrying zinc oxide nanoparticles against infection and inflammation, and to evaluate its effect on wound healing. METHODS:Thirty-two Sprague-Dawley rats with a full-thickness wound on the back skin were randomly divided into two groups. In experimental groiup, porous collagen/silk fibroin scaffolds containing zinc oxide nanoparticles were implanted, while only collagen/silk fibroin scaffolds were implanted in control group. Wound healing was compared between the two groups by measuring residual wound area at 1, 2, 4, 8 weeks post implantation. Hematoxylin-eosin and interleukin 6 immumohistochemical staining were performed at 1, 2, 4 weeks post implantation to observe wound morphology and inflammatory reactions. Meanwhile, expression of interleukin 6 and interleukin 1β was detected by real-time PCR. RESULTS AND CONCLUSION:(1) At 2, 4, 8 weeks post implantation, significantly increased healing rate was observed in the experiment group compared with the control group (P<0.05). (2) Findings from the hematoxylin-eosin staining showed that obvious inflammatory cell infiltration was observed in the control group, but less inflammation with vigorous growth of granulation tissues on the wound surface occurred in the experimental group at 1 week after implantation. Then, the wound repair was basically completed in the experimental group presenting with complete and compact epidermal tissue structure, while scar formation with no skin cover was found in the control group at 4 weeks after implantation. (3) Findings from the interleukin 6 immumohistochemical staining showed that there was interleukin 6 positive expression in both two groups to different extents; at 4 weeks after implantation, the expression of interleukin 6 was remarkably reduced in the control group, but it was still a strong positive expression, while week positive expression of interleukin 6 was observed in the experimental group. (4) Compared with the control group, the mRNA expression of interleukin 6 and interleukin 1β was both lower in the experimental group at 1, 2, 4 weeks after implantation, but there was a significant difference between the two groups at 1 and 2 weeks after implantation (P<0.05). Overall, the porous collagen/silk fibroin scaffold carrying zinc oxide nanoparticles can effectively reduce inflammations following skin injury, and accelerate skin wound healing.