Protective effect of betaine against lead-induced testicular toxicity in male mice.

Lead (Pb) is an environmental toxicant reported to impair male reproductive system. Betaine is a natural product which has promising beneficial effects against oxidative stress. In this experimental study, we evaluated the ameliorative effect of betaine on sperm quality and oxidative stress induced by lead (Pb) in the testis of adult male mice. Sixty male Kunming mice were divided equally into four groups: control group, betaine group (1% in drinking water), lead group (100 mg kg-1  bw-1  day-1 ) and betaine + lead group. In the last group, mice were supplemented with betaine for two weeks prior to the initiation of lead treatment and concurrently during lead treatment for 3 weeks until sacrificed. Our results indicated that in the lead-administrated group, body weights together with sperm count were significantly decreased (p < .05). The numbers of abnormal sperms were found to be higher in lead-treated mice. The activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and catalase (Cat) were significantly reduced, while the level of malondialdehyde (MDA) content was increased in the testis tissue following lead treatment. The mRNA levels of antioxidant-related genes (SOD1, GPX1 and CAT) were significantly decreased in the lead group. Betaine enhanced these parameters in betaine + lead group. In testis histology span, Johnson score was decreased (p < .05) in lead group and co-treatment with betaine increased Johnson score significantly in betaine + lead group. These results indicate that betaine improves sperm quality and ameliorate oxidative damage in testis of mice exposed to lead.

Heme oxygenase 1 regulates apoptosis induced by heat stress in bovine ovarian granulosa cells via the ERK1/2 pathway.

Heat stress can inhibit follicular development in dairy cows, and thus can affect their reproductive performance. Follicular granulosa cells can synthesize estrogen, that affects the development and differentiation of follicles by apoptosis. Heme oxygenase 1 (HO-1/heat shock protein 32) plays an antiapoptotic and cytoprotective role in various cells during stress-induced apoptosis, but little is known about its definitive function in bovine (ovarian) granulosa cells (bGCs). In our study, the roles and mechanism of HO-1 on the heat stress-induced apoptosis of bGCs were studied. Our results show that the expression of HO-1 was significantly increased under heat stress. Moreover, HO-1 silencing increased apoptosis, whereas its overexpression dampened apoptosis by regulating the expression of Bax/Bcl-2 and the levels of cleaved caspase-3. In addition, HO-1 can also play a cytoprotective role by affecting estrogen levels and decomposing heme to produce biologically active metabolite carbon monoxide (CO). Meanwhile, CO significantly increased the level of HO-1, decreased Bax/Bcl-2 levels, and inhibited the activation of extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway. The apoptosis of ovarian GCs can affect the secretion of estrogen and lead to disorder of the ovarian microenvironment, thus affecting the normal function of the ovary. Our results indicate that HO-1 acts as a cytoprotective enzyme and plays a protective role in heat-induced apoptosis of bGCs. In conclusion, HO-1 and its metabolite CO inhibit the apoptosis of bGCs induced by heat stress through the ERK1/2 pathway. The results of this study provide a valuable clue for improving the fertility of heat stressed cows in summer.

SIRT7 depletion inhibits cell proliferation, migration, and increases drug sensitivity by activating p38MAPK in breast cancer cells.

SIRT7 is a member of the sirtuin family of proteins that are known to be associated with tumor development. However, the functional roles and molecular mechanisms underlying the function of SIRT7 in breast cancer cell survival and tumor development remain unclear. Recent studies demonstrated that SIRT7 is upregulated in breast cancer cells and tissues. In the present study, we systematically explored the roles of SIRT7 in the growth of breast cancer cells and tumors both in vitro and in vivo. Our results showed that SIRT7 plays a major role in facilitating cell survival by promoting cell proliferation and inhibiting apoptosis. SIRT7 depletion significantly inhibited cell invasion and wound healing by blocking cell cycle progression and inducing cell apoptosis. Meanwhile, SIRT7 depletion can increase the sensitivity of breast cancer cells to doxorubicin (DOX). Xenograft model studies showed that stable silencing of SIRT7 inhibited tumor growth and enhanced tumor sensitivity to DOX. Further research revealed that p38MAPK is involved in SIRT7-mediated regulation of breast cancer cell proliferation and tumor growth. Taken together, our results showed that SIRT7 plays a critical role in breast cancer cell survival, migration, and tumor growth, and increased the efficiency of DOX treatment both in vitro and in vivo. Therefore, SIRT7 is a promising therapeutic target in breast cancer treatment.

Heat stress impairs mice granulosa cell function by diminishing steroids production and inducing apoptosis.

Ovarian injury can be induced by heat stress. Mice granulosa cells (GCs) are critical for normal ovarian function and they synthesize a variety of growth factors and steroids for the follicle. Furthermore, the growth, differentiation, and maturate of theca cells and oocyte are dependent upon the synthesis of GCs. Due to the critical biological functions of GCs, we hypothesized that the apoptosis and dysfunction of GCs could also be induced by heat stress. We analyzed GCs apoptosis and evaluated the expression of apoptosis-related genes (caspase-3, Bax, Bcl-2) after heat treatment. Radio immunity assay was used to measure the secretion of 17ß-estradiol (E2) and progesterone (P4). RT-PCR was used to evaluate the expression of steroids-related genes (Star, CYP11A1, CYP19A1). Our data suggested that heat stress inhibited GCs proliferation, induced GCs apoptosis, decreased E2 and P4 secretion, reduced the steroids-related genes mRNA expression. Besides, our results indicated that heat treatment-induced apoptosis of GCs through the mitochondrial pathway, which involved caspase-3 and Bax. The reduction in steroids secretion and mRNA expression of Star, CYP11A1, and CYP19A1 might also play a role in heat-induced GCs apoptosis and ovarian injury.

Heat shock protein 32/heme oxygenase-1 protects mouse Sertoli cells from hyperthermia-induced apoptosis by CO activation of sGC signalling pathways.

Heat shock protein 32 (Hsp32)/heme oxygenase-1 (HO-1) may be a key enzyme for the protection of cells against stress. Its anti-apoptotic effect has been attributed to its product, carbon monoxide (CO), in many types of cells. However, whether its anti-apoptotic mechanism plays a role in Sertoli cells (SCs) is not yet clear. We hypothesise that Hsp32/HO-1 and CO generated from it provide survival advantages in SCs by preventing apoptosis under heat exposure. After treatment of cultured SCs with hyperthermia and/or Hsp32/HO-1 activater hemin, apoptosis was measured valuated by annexin V-FITC and caspase-3 activation. We have also analysed the Hsp32/HO-1-derived CO content of cultured media and cyclic guanosine monophosphate (cGMP) production by enzyme-linked immunosorbent assay (ELISA). Hyperthermia induced SCs apoptosis, while preincubation with hemin suppressed SC hyperthermia-induced apoptosis. Hyperthermia and/or hemin increase Hsp32/HO-1 gene expression and the production of CO, which, in turn, stimulates the generation of cGMP. The results suggest that Hsp32/HO-1 is a protective factor in heat-stressed SCs, and that CO generated from Hsp32/HO-1 is involved in the anti-apoptotic pathway.

Percutaneous kyphoplasty in the treatment of Kümmell disease in lumbar scoliosis: A case report.

BACKGROUND: Due to mechanical imbalance in the spine, elderly scoliosis patients tend to develop vertebral fracture nonunion, i.e., Kümmell disease, when osteoporotic vertebral compression fractures occur. However, accompanying vertebral rotational deformities make surgical procedures challenging risky. Such patients are usually compelled to undergo conservative treatment and there are very few reports on minimally invasive surgeries for them. We first-time report a patient with Kümmell disease and lumbar scoliosis treated with percutaneous kyphoplasty (PKP) under O-arm guidance. CASE SUMMARY: An 89-year-old female was admitted to the hospital due to delayed low back pain after a fall. She was diagnosed with Kümmell disease based on physical and radiologic examinations. The patient experienced severe scoliosis and subsequently underwent O-arm-guided kyphoplasty, resulting in a significant alleviation of low back pain. CONCLUSION: PKP has good efficacy in treating Kümmell disease. However, surgical risks are elevated in scoliosis patients with Kümmell disease due to the abnormal anatomical structure of the spine. O-arm assisted operations play a crucial role in decreasing surgical risks.

Conservation tillage facilitates the accumulation of soil organic carbon fractions by affecting the microbial community in an eolian sandy soil.

Conservation tillage (CT) is an important agronomic measure that facilitates soil organic carbon (SOC) accumulation by reducing soil disturbance and plant residue mulching, thus increasing crop yields, improving soil fertility and achieving C neutrality. However, our understanding of the microbial mechanism underlying SOC fraction accumulation under different tillage practices is still lacking. Here, a 6-year in situ field experiment was carried out to explore the effects of CT and traditional tillage (CK) practices on SOC fractions in an eolian sandy soil. Compared with CK, CT increased the particulate OC (POC) content in the 0-30 cm soil layer and the mineral-associated OC (MAOC) content in the 0-20 cm soil layer. Moreover, tillage type and soil depth had significant influences on the bacterial, fungal and protistan community compositions and structures. The co-occurrence network was divided into 4 ecological modules, and module 1 exhibited significant correlations with the POC and MOC contents. After determining their topological roles, we identified the keystone taxa in the network. The results indicated that the most common bacterial taxa may result in SOC loss due to low C use efficiency, while specific fungal (Cephalotrichum) and protistan (Cercozoa) species could facilitate SOC fraction accumulation by promoting macroaggregate formation and predation. Therefore, the increase in keystone fungi and protists, as well as the reduction in bacteria, drove module 1 community function, which in turn promoted SOC sequestration under CT. These results strengthen our understanding of microbial functions in the accrual of SOC fractions, which contributes to the development of conservation agriculture on the Northeast China Plain.

The Signaling Mechanism of TGF-ß1 Induced Bovine Mammary Epithelial Cell Apoptosis.

The present study showed that Transforming growth factor beta 1 (TGF-ß1) can induce apoptosis of bovine mammary epithelial cells. This apoptosis was also observed with phosphorylation of Smad2/3 within 0.5-2 h. Afterwards the signal transferred into the nucleus. Moreover, intracellular free Ca(2+) concentration was significantly elevated as well as Caspase-3 activated and DNA lysised, thereby inducing the programmed cell death. This signaling pathway of TGF-ß1 was blocked by SB-431542 (10(-2) µM) via inhibiting ALK-5 kinase activity, which thus reversed the anti-proliferation and apoptosis effect of TGF-ß1 in mammary epithelial cells. These results indicated that TGF-ß1 induced apoptosis of bovine mammary epithelial cells through the ALK-5-Smad2/3 pathway, which plays an important role in inhibiting survival of mammary epithelial cells. Moreover, intracellular Ca(2+) also played a critical role in TGF-ß1-induced cell apoptosis.

Expression of vascular endothelial growth factor and matrix metalloproteinase-9 in sacral chordoma.

Sacral chordoma is a vessel-rich and infiltrative tumor, but the fundamental knowledge of its biological behavior remains unknown. This study was designed to investigate the expression levels and contributions of vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) in the angiogenesis and recurrence of sacral chordoma and their correlations. An immunohistochemical method was used to investigate the expression of VEGF, MMP-9, and microvascular density (MVD) in 36 patients with sacral chordoma. Their differences in expressions were statistically analyzed and their correlations with angiogenesis and recurrence were evaluated. The mean MVD of sacral chordomas was significantly higher than that of the adjacent normal tissues (P = 0.033). Immunoreactivity for VEGF and MMP-9 was significantly higher in sacral chordoma tissues than in adjacent normal tissues (P = 0.008, P = 0.005). The mean MVD of VEGF and MMP-9 were statistically higher in positive group than in negative group (P = 0.015, P = 0.004), respectively . Moreover, a significant correlation was found between the VEGF and MMP-9 (P = 0.002). The log-rank test revealed that continuous disease-free survival time (CDFS) was significantly shorter in the MMP-9-positive group than in the MMP-9-negative group (P = 0.019), but the difference in the VEGF-positive group and the VEGF-negative group was not statistically significant (P = 0.938). Our data suggest that VEGF and MMP-9 might act with a synergistic effect and can positively regulate the angiogenesis in sacral chordoma. Positive expression of MMP-9 might indicate the local recurrence of sacral chordoma. The result suggests that some specific drugs which inhibit VEGF, MMP-9, or their receptors may have a good therapeutic effect for sacral chordoma.

Risk factors for postoperative wound infections of sacral chordoma after surgical excision.

STUDY DESIGN: A retrospective study, analyzing the risk factors for postoperative wound infections of the sacral chordoma after surgical excision. OBJECTIVE: To determine the preoperative, intraoperative, and patient characteristics that contribute to an increased risk of postoperative wound infection in patients undergoing sacral chordoma resection. SUMMARY OF BACKGROUND DATA: Postoperative wound infection after spinal operations is a dreaded complication. The risk factors have been investigated earlier, but the patients with sacral chordoma may be distinct. METHODS: Between January 1992 and December 2007, 45 patients with sacral chordomas were treated with surgical resection. Data regarding preoperative and intraoperative risk factors for postoperative wound infection were evaluated using univariate analysis and multivariable conditional logistic regression. Odds ratios with 95% confidence intervals and P values were calculated. RESULTS: Of the 45 patients with sacral chordoma, 16 (35.6%) acquired postoperative wound infection. Significant risk factors associated with postoperative wound infection in the univariate analysis included the following: albumin <3.0, previous surgery, operating time, instrumentation, and surgical team. Albumin<3.0, operating time >6 hours, and previous surgery were statistically significant in the multivariable model. CONCLUSIONS: Patients undergoing sacral tumor surgery may be at greater risk for developing wound complications. In this study, it seems that albumin<3.0, operating time >6 hours, and previous surgery may predict those patients that were more prone to developing postoperative wound infection. Using a single surgical team and no instrumentation seems to provide protection against postoperative wound infection in this patient population.

[Relationships of three microsatellite loci with GSH-Px, SOD and Na+/K+-ATP enzyme activities and daily milk yield in different seasons in Holstein cows].

Genetic variation of three microsatellite loci BMS2258, SOD1, and BM723, which were closely correlated with GSH-Px, SOD, and Na+/K+-ATPase genes, was analyzed in 130 Holstein cows by PCR and nondenaturing polyacrylamide gel-electrophoresis. Polymorphic information content, effective number of alleles and heterozygosity of these microsatellite loci were determined. Relationships of the three microsatellite loci with enzyme activities and daily milk yields in Holstein cows were analyzed by least squares linear model. The results showed significant correlations of the three microsatellite loci with their corresponding enzyme activities and daily milk yield in summer and fall (Plt;0.05). The least square means of GSH-Px activities and daily milk yields for BMS2258 (182 bp/164 bp), SOD activities for SOD1 (148 bp/148 bp), and daily milk yields for SOD1 (148 bp/146 bp), Na+/K+-ATPase activities and daily milk yields for BM723 (161 bp/111 bp) were relatively higher. These genotypes were the most favorable genotypes for enzyme activity and daily milk yields in summer and fall, which could be references for marker assisted selection in heat resistance traits in dairy cattle.

[Analysis of HSP70 mRNA level and association between linked microsatellite loci and heat tolerance traits in dairy cows].

The objective of this study was to investigate the variation of HSP70 mRNA level in dairy cows and relationships of its closely linked microsatellite loci with heat tolerance traits. Blood samples were collected from ten healthy Holstein cows with the same age and milking stage at different temperatures-humid-index (THI) (86.2, high temperature; 70.9, critical high temperature, and 56.8, optimum temperature). The mRNA levels of HSP70 of lymphocytes in peripheral blood were analyzed using real-time RT-PCR. The mRNA level of HSP70 was increased with the THI; the mRNA level of HSP70 at high temperature was higher than others (P<0.01). This indicated that the bovine HSP70 gene may act as a potential can-didate gene for response to heat shock. Genetic variation of three microsatellite loci BMS468, BM1258, and BM1815, which were closely linked to HSP70 gene on chromosome 23, was analyzed in 160 Holstein cows with non-denaturing poly-acrylamide gel electrophoresis. The association between these microsatellite loci and heat tolerance traits were analyzed by least square linear model. The results showed that 134 bp/128 bp at BMS468 and 186 bp/148 bp at BM1815 were the most favorable genotypes for HTC, red cell potassium, and decrement rate of milk yield in high temperature (P<0.05); 101 bp/99 bp at BM1258 was the most favorable genotype for decrement rate of milk yield in high temperature (P<0.05).

[Balloon kyphoplasty for osteoporotic vertebral compression fractures with osteonecrosis].

OBJECTIVE: To explore the clinical efficacy of the percutaneous balloon kyphoplasty for osteoporotic vertebral compression fractures with osteonecrosis. METHODS: The clinical data of 31 patients with osteoporotic vertebral compression fractures associated with osteonecrosis from January 2005 to January 2008 were analyzed retrospectively. There were 13 male and 18 female in this study. The mean age of the patients was 71 years (range from 57 to 84 years). The back pain lasted for 4.2 months (from 1 month to 10 years). Radiography, MRI and CT examination were performed. The patients were treated by percutaneous balloon kyphoplasty and the vertebral body tissue was extracted to perform common pathological examination. The anterior vertebral height was measured on a standing lateral radiograph before operation, after operation (one day after operation) and at the final follow-up. A Visual Analog Scale (VAS) and the Oswestry Disability Index (ODI) were chosen to evaluate pain status and functional activity. RESULTS: The mean follow-up was for 27 months (range, 18 to 48 months). The anterior vertebral body height of fracture vertebra was restored from (34.7 +/- 3.1)% preoperatively to (71.4 +/- 2.3)% postoperatively, and to (70.2 +/- 2.5)% at the final follow-up. There was a significant improvement between preoperative and postoperative values (P < 0.05) and no difference between postoperatively and at the final follow-up (P > 0.05). The VAS was 8.7 +/- 0.4 preoperatively, 2.3 +/- 0.7 postoperatively, and 1.9 +/- 0.2 at the final follow-up; and the ODI was 89.1 +/- 2.7 preoperatively, 31.7 +/- 3.1 postoperatively, and 29.1 +/- 2.7 at the final follow-up. There was statistically significant increment in the VAS and ODI postoperatively compared with preoperatively (P < 0.05), while there was no statistically significant differences between postoperatively and at the final follow-up (P > 0.05). There was a significant increment between preoperative and final follow-up values (P < 0.05). Asymptomatic cement leakage occurred in two cases. New vertebral fracture occurred in one case. CONCLUSION: Balloon kyphoplasty is a safe and effective procedure for osteoporotic vertebral compression fractures with osteonecrosis.

Surgery for thoracic myelopathy caused by ossification of the ligamentum flavum.

Between January 1996 and December 2003, our department treated 16 patients (10 men and 6 women; average age 57.5 years) by performing a laminectomy for thoracic myelopathy caused by ossification of the ligamentum flavum (OLF). We followed up all patients for 36 to 86 months (mean follow-up time, 57.3 months). The mean (+/-standard deviation) Japanese Orthopaedic Association score increased from 5.0+/-1.4 points before the operation to 7.7+/-1.9 points at the last follow-up (p<0.01). The average values for pre-operative and post-operative kyphosis of the involved vertebrae were 5.8 degrees +/-4.1 degrees and 8.8 degrees +/-6.0 degrees , respectively; the mean increase in kyphosis was only 3.0 degrees +/-2.4 degrees . An intraoperative dural tear was the main complication and none of the patients developed severe neurological complications. We conclude that laminectomy was both effective and safe in the treatment of thoracic OLF, but it must be performed with great care because of frequent dural adhesions to the OLF. The increase in kyphosis after the laminectomy was minimal when most of the facet joints were left intact and when the patient followed a back extensor exercise program post-operatively.

SIRT7 Regulates Lipopolysaccharide-Induced Inflammatory Injury by Suppressing the NF-κB Signaling Pathway.

Mastitis has severely affected the cattle industry worldwide and has resulted in decreased dairy production and cattle reproduction. Although prevention and treatment methods have been implemented for decades, cattle mastitis is still an intractable disease. Sirtuin 7 (SIRT7) is an NAD+-dependent deacetylase that is involved in various biological processes, including ribosomal RNA synthesis and protein synthesis, DNA damage response, metabolism, and tumorigenesis. However, whether SIRT7 participates in inflammation remains unknown. Our results revealed that SIRT7 is downregulated in tissue samples from mastitic cattle. Therefore, we isolated dairy cow mammary epithelial cells (DCMECs) from breast tissues and developed an in vitro model of lipopolysaccharide- (LPS-) induced inflammation to examine SIRT7 function and its potential role in inflammation. We showed that SIRT7 was significantly downregulated in LPS-treated DCMECs. SIRT7 knockdown significantly increased the LPS-stimulated production of inflammatory mediators, like reactive oxygen and nitric oxide, and upregulated TAB1 and TLR4. In addition, SIRT7 knockdown significantly increased the phosphorylation of TAK1 and NF-κBp65 in LPS-treated DCMECs. Moreover, SIRT7 knockdown promoted the translocation of NF-κBp-p65 to the cell nucleus and then increased the secretion of inflammatory cytokines (IL-1ß and IL-6). In contrast, SIRT7 overexpression had the opposite effects when compared to SIRT7 knockdown in LPS-treated DCMECs. In addition, SIRT7 overexpression attenuated LPS-induced DCMEC apoptosis. Taken together, our results indicate that SIRT7 can suppress LPS-induced inflammation and apoptosis via the NF-κB signaling pathway. Therefore, SIRT7 may be considered as a potential pharmacological target for clinical mastitis therapy.

[Analysis of the kinematic parameters of hyperactivated Guinea pig spermatozoa].

OBJECTIVE: To evaluate the role of sperm kinematic parameters in the hyperactivation of Guinea pig spermatozoa, and to confirm the index of their hyperactivated motility. METHODS: Computer-aided sperm analysis (CASA) was used to describe the kinesis parameters of the Guinea pig spermatozoa incubated for 1, 3, 5 and 7 hours. RESULTS: The curvilinear velocity, average path velocity and amplitude of lateral head displacement were increased with time and reached the peak at 5 hours, while the straight linear velocity, linearity, straightness and beat cross frequency were gradually decreased with time and hit the bottom at 5 hours. CONCLUSION: The sperm movement pattern changes greatly before hyperactivation during the capacitation of Guinea pig spermatozoa.

[Diagnosis and surgical treatment of thoracolumbar vertebral Chance fractures caused by a fall].

OBJECTIVE: To explore the diagnosis of Chance fractures of thoracolumbar spine and the clinical efficacy of segmental pedicle screw instrumentation in the treatment. METHODS: The clinical data of 16 patients with Chance fractures from January 2000 to January 2007 were retrospectively analyzed. All patients were treated with segmental pedicle screw instrumentation. Radiology and the Oswestry Disability Index (ODI) were used to evaluate the efficacy. RESULTS: One case associated with open dislocation of ankle and craniocerebral injury, 3 cases with calcaneus fracture and 1 with kidney contusion. All cases were followed up from 7 months to 6 years, with a mean of 2.6 years. There were no operation complications. The anterior vertebral body height of fracture vertebra was restored from preoperative (63.3 +/- 6.8)% to postoperative (92.1 +/- 4.0)%, at the follow-up (90.7 +/- 3.6)%. There was a significant improvement between preoperative and postoperative values (P < 0.01) and no difference between initial postoperative and final follow-up (P > 0.05). The mean ODI was 4.9. CONCLUSIONS: The injuries associated with Chance fracture caused by a fall and its mechanism are different from those with the safety belt injury. The segmental pedicle screw instrumentation can rebuild spinal stabilization and restore spinal alignment in treatment of Chance fractures.

[Determination of painful vertebrae treated by kyphoplasty in multiple-level vertebral compression fractures].

OBJECTIVE: To explore how to determine painful vertebrae treated by kyphoplasty in multiple-level osteoporotic vertebral compression fractures and clinical outcome. METHODS: From October 2002 to June 2005, 51 consecutive procedures with kyphoplasty were performed on 35 patients with multiple-level osteoporotic vertebral compression fractures. There were 51 painful vertebrae among 120 vertebral compression fractures. The painful vertebra was determined by the signal intensity changes in MR images, combined with radiography and local percussion pain before operation. Only painful vertebrae were treated by kyphoplasty. Preoperative, postoperative and final follow-up visual analog scale (VAS) and radiographic findings such as vertebral height and Cobb angle were recorded and analyzed. RESULTS: All patients tolerated the procedure well with immediate relief of their back pain after kyphoplasty and they can walk at 1-3 days after the procedure. There were 3 vertebrae (3/51) occurred asymptomatic extravasation of cement. 31 cases were followed up for mean 16.2 months (range 6-44 months). VAS reduced from preoperative 8.7 to final follow-up 2.1 (P <0. 01). At final follow-up the vertebral height had a recovery rate of 59.17%, and the mean Cobb angle was improved 10.1 degrees. There was a significant improvement between preoperative and final follow-up values (P < 0.01). CONCLUSIONS: The painful vertebra can be determined by signal intensity changes in MR series images in multiple-level osteoporotic vertebral compression fractures. Selecting painful vertebrae to be treated by kyphoplasty can make patients with multiple-level VCFs gain an excellent result.

[Effects of different donor cells on the development of nuclear-transferred porcine embryos].

This study was undertaken to systematically examine the effects of different donor cells and numbers of passages on the development of nuclear-transferred porcine embryos, so as to establish a preliminary procedure for porcine cloning. Porcine oocytes obtained at slaughter were matured in vitro for 40-44 h and then enucleated in manipulation medium containing 5 micro g/mL cytochalasin B. Fibroblast cells (FC), oviduct epithelial cells (OEC), granulosa cells (GC) and cumulus cells (CC) after 3-9 passages in 10% FBS-supplemented culture medium were either treated by serum starvation (0.5% FBS for 2-9 days), 0.1 micro g/mL aphidiconlin (APD) for 1 day and 0.5% FBS for 2-9 days or left untreated in complete medium for 2-9 days. They were transferred into enucleated oocytes by microinjection or electric fusion (100 V/mm, 30 ms and 1 pulse). Reconstituted embryos were activated with a combination of calcium ionophore A23187 or electric pulse and 6-DMAP, and cultured for 6 days, to evaluate their cleavage and embryonic development. The cleavage rate of embryos reconstructed with FC and GC pretreated with 0.1 micro g/mL APD + 0.5% FBS were significantly higher than that of serum starvation group and control group (P<0.01). There was a significant difference in the cleavage rate and embryonic development among embryos derived from GC, CC and FC, OEC pretreated with 0.1 micro g/mL APD + 0.5% FBS. The cleavage rate of embryos reconstructed with GC by electrofusion was significantly higher than that by microinjection (P<0.05), but no difference was found in the proportion of embryos that developed to blastocysts. About 75% to 85% of GC at 3 and 6 passages, and FC at 6 and 10 passages had a normal karyotype, and resulted in similar cleavage rate and blastocyst development. These results indicate that: (1) FC and GC can be cultured up to 9 passages and maintain a relatively stable karyotype; (2) Treatment of donor cells with 0.1 micro g/mL APD prior to nuclear transfer can improve the efficiency of somatic cell nuclear transfer in buffalo but serum starvation is inefficient in our system; (3) Both FC and GC cells can be used as the donor karyoplasts for nuclear transfer, and their efficiency is not influenced by the culture passages. (4) The development of reconstructed embryos by electrofusion is higher than that by microinjection, but there is no difference in the overall efficiency between the two methods.

[CatSper and sperm hyperactivation].

Elevation of sperm Ca2+ seems to be responsible for an asymmetric form of motility called hyperactivation, which is first seen near the time of fertilization. CatSper family proteins are putative cation channels expressed exclusively in the membranes of the sperm flagellum. Hyperactivation requires CatSper proteins, which presumably serve as the route of entry for Ca2+ that operates directly on the flagellar axoneme to increase waveform asymmetry. In this article, the structure of mouse CatSper, the role it plays in sperm hyperactivation and the unsolved problems are described.