OBJECTIVE: Inter-oocyte cytoplasmic transfer has been performed recently as a means to improve the outcome of assisted reproductive methods and becomes a hotspot. To overview the recent advances and problems in this area in this article, and put forth a new concept of reverse cloning technique based on ooplasmic transfer. DATA SOURCES: An online search of PUBMED was undertaken to identify correlative English articles published from January 1998 to December 2006 by using the keywords of "ooplasmic transfer, mitochondria heteroplasmy,animal cloning". Meanwhile, relevant Chinese articles published between January 1998 and December 2005 were searched in Wanfang database with the same keywords in Chinese. STUDY SELECTION:The articles were primarily screened, and the citations at the end of each reference were reviewed. Inclusion criteria: The articles related to the research of ooplasmic transfer, cloning or reverse cloning. Exclusion criteria: Repetitive studies or Meta analytical articles. DATA EXTRACTION:Totally 126 relevant articles were collected, and 31 of them were in accordance with the inclusion criteria, while 95 of them were excluded because of repetition or obsolescence. In 31 articles met the criteria, 17 studies were relevant to the ooplasmic transfer, 13 studies related to animal cloning and one study related to reverse cloning. DATA SYNTHESIS: Exciting advances have been achieved in human gamete biology, and ooplasmic transfer have been widely applied. Many studies indicated that mitochondria in oocytoplasm affected obviously the fertilization of the oocytes and early embryo development. Lots of experiments have been done on the interaction of cytoplasm and nucleus as well as the mechanism of genomic reprogramming to further the investigation. Meanwhile, the techniques are progressed. Cloning techniques is excellent in application prospect, while there are some problems at the same time. Therefore the concept of reverse cloning technique was proposed based on ooplasmic transfer. CONCLUSION: The mitochondria in oocytoplasm has remarkable effect on the fertilization of the oocytes and early embryonic development. However, ooplasm transfer can lead to mitochondria DNA heteroplasmy, and more work must be done on potential problems. The reverse cloning technique based on the ooplasmic transfer has a good prospect.

Objective To observe the effects of cystamine, a tissue transglutaminase (tTG) inhibitor, on the development of rat liver fibrosis induced by carbon tetrachloride. Methods One hundred male SD rats were randomly divided into control group (n=20), hepatic fibrosis group (n=40) and cystamine group (n=40) . Liver fibrosis model was induced by intraperitoneal injection of carbon tetrachloride. Cystamine (112 mg·kg-1·d-1) was administered by intraperitoneal injection 2 days before injection of carbon tetrachloride. The rats were sacrificed at weeks 4 and 8, and the liver tissues and serum specimens were obtained. The mRNA expression of tTG, smooth muscle-alpha (α-SMA), collagen-Ⅰ and tissue inhibitors of metalloproteinase-1 (TIMP-1) were detected by real time PCR. The protein expression of tTG and α-SMA, liver function and content of hydroxyproline in liver tissues were determined by Western blot. Histological changes of the liver was observed under microscope. The fibrosis conditions of rat liver in each group were evaluated according to the semi-quantita-tive scoring system. All the data were analyzed by one-way ANOVA. Results Eight weeks after the injection of carbon tetrachloride, obvious injury of the liver in liver fibrosis group was observed. The levels of alanine trans-aminase (ALT), total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (1313±157)U/L, (99.9±18.5)μmol/L, (10.9±1.6)μmoL/L, (55±12)μg/g, 145.6±51.2, 130.3±44.6, 211.3±75.1 and 162.4±53.5. After administration of cystamine, the levels of ALT, total bile acid, total bilirubin, hydroxyproline, tTG, α-SMA, collagen-Ⅰ and TIMP-1 were (378±87) U/L, (61.0±12.7) μmol/L, (9.8±1.7) μmol/L, (70±14 ) μg/g, 48.6±12.3, 40.7±12.3, 63.9±16.0, 59.2μ23.4. Conclusion Cystamine can alleviate the carbon tetrachloride-induced rat liver fibrosis by inhibiting the tTG pathway.

Acute Disease ; Adult ; Carbon Tetrachloride Poisoning ; diagnosis ; drug therapy ; Humans ; Male

Objective To investigate the distribution of hepatitis B virus (HBV) genotypes in patients with chronic HBV infections and the correlations of genotypes with liver fibrosis and hepatocellular carcinoma (HCC).Methods Serological,virological and pathological data of 190 patients with chronic HBV infections admitted to Hangzhou First People's Hospital during June 2007 and June 2012 were retrospectively analyzed.The series included 62 cases of chronic hepatitis B (CHB),60 cases of liver cirrhosis and 68 cases of HCC.HBV was genotyped by multiplex PCR,and subgenotyped by restriction fragment length polymorphism (RFLP).SPSS 11.0 was used for statistical analysis.Results Among 190 patients,61 were HBV genotype B (32.1％),126 were genotype C (66.3％),and 3 were B + C hybrid.HBV B2 (61/61,100.0％) and C2 (123/126,97.6％) were the major subgenotypes.HBV genotype B was more prevalent in CHB patients (46.8％,29/62) than in liver cirrhosis (20.0％,12/60) and in HCC patients (29.4％,20/68) (x2 =8.73 and 4.16,P＜0.01 or P＜0.05),whereas the prevalence of genotype C was higher in liver cirrhosis and HCC than that in CHB (x2 =9.54 and 4.17,P ＜0.01 or P ＜ 0.05).Patients with genotype C2 had higher serum hyaluronic acid level than with genotype B2 in 3 groups (t =2.685,2.433 and 2.015,P ＜ 0.01 or P ＜ 0.05).CHB patients with C2 infections had higher liver fibrosis grade than those with B2 (x2 =6.726,P =0.010),while there was no statistical difference in liver inflammation grade (x2 =0.601,P ＞ 0.05).HCC patients with B2 infection tended to have larger tumor diameter (≥5 cm) (x2 =7.231,P ＜ 0.01) and those with C2 infection were prone to be more serious cirrhosis (x2 =4.910,P ＜ 0.05).Conclusions HBV genotypes C2 and B2 are predominant in patients with chronic HBV infections in Hangzhou.HCC patients infected with HBV C2 may be complicated with more severe liver fibrosis,and those with HBV B2 infections may tend to have larger liver tumor.

Objective To explore the efficacy of tepronone and folic acid in the treatment of chronic atrophic gastritis (CAG) evaluated by the marking targeting biopsy (MTB).Methods A total of 224 H.pylori negative CAG patients were selected and divided into group A (n 96,tepronone 50 mg/time,folic acid 10 mg/time,three times/day),group B (n=23,tepronone 50 mg/time,three times/day),group C (n=74,unspecific treatment) and group D (n=31,no treatment).The treatment course lasted for one year.The clinical symptoms improvement of each group was observed before and after treatment.The pathological improvement of gastric mucosa by MTB was inspected before and after treatment.The chi square test was performed for the comparison between groups.Results The total efficacy rates of group A,B,C and D were 43.8％ (42/96),39.1％ (9/23),33.8％(25/74) and 32.3％ (10/31) respectively,there was no significant difference between groups (x2 =2.328,P =0.507).For the significant efficacy rate of gastric mucosa pathological improvement,group A was compared with group D,group A was compared with group C and group B was comparedwith group D,the differences were significant (x2 =14.520,14.628 and 8.995,all P＜0.01).In the total efficacy rate of gastric mucosa pathological improvement,group A (49.8％,131/263) was compared with group D (24.2％,16/66),group A was compared with group C (35.9％,66/184)and group B (44.7％,21/47) was compared with group D,the differences were significant (x2 =13.953,8.535 and 5.207,all P＜0.05).Conclusion Teprenone alone or teprenone and folic acid combination can obviously improve pathological changes of CAG patients.

BACKGROUND: Chinese medicine Gukang prescription has a clear effect on clinical treatment of osteoporosis, but the therapeutic pathway is stil unclear.
OBJECTIVE:To investigate the effects of Chinese medicine Gukang on the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin by regulating core binding factor alpha 1 expression to control the growth and development of osteoblasts.
METHODS:Sprague-Dawley neonatal rats within 24 hours after delivery were used for the separation and culture of osteoblasts. Adult Sprague-Dawley rats were used to prepare drug-containing serum, and then divided into two groups randomly:normal control group and Gukang group. Rats in the normal control and Gukang groups were intragastrical y administrated with extract of Gukang prescription and normal saline based on rat’s body surface area, for 1 consecutive week. Two hours after the last administration, blood samples were taken from the heart. Then the serum was col ected. Osteoblasts at passage 3 were confirmed with alkaline phosphatase assay and digested. After counting and planting, al osteoblasts were divided into two groups and treated with col ected
serum for 72 hours. Proliferative rate of osteoblasts was detected by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test. Secretion of alkaline phosphatase was detected by using enzyme linked immunosorbent assay and corrected with the corresponding absorbance value. mRNA expression of core binding factor alpha 1, receptor activator of nuclear factor kappa B ligand and osteoprotegerin were detected by using reverse
transcription-PCR in al groups.
RESULTS AND CONCLUSION:mRNA expression of osteoprotegerin and core binding factor alpha 1 in the Gukang group was significantly higher than that in the normal control group, but protein and mRNA expression of receptor
activator of nuclear factor kappa B ligand were dramatical y lower in the Gukang group compared with the normal control group (P<0.01). These findings indicate that Chinese medicine Gukang prescription can modulate the expression of
core binding factor alpha 1, thereby adjusting the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin, which may be one of the mechanisms underlying Gukang treatment for osteoporosis.

Objective To investigate the anti-fibrosis effect of a 5-lipoxygenase (5-LO)specific inhibitor AA-861 on liver fibrosis. Methods Liver fibrosis was induced in male Sprague Dawley rats by intraperitoneal injection of carbon tetrachloride(CCI4).AA-861(0.2 mg/100 g/d)was administrated by intraperitoneal injection starting 2 days before the first dosage of CCI4 and rats were killed at weeks 2,4,and 6.Liver specimens were obtained from each animal and fixed with 4%formaldehyde for histological analysis. The Mrna expression of 5-LO,smooth muscle-alpha(α-SMA),Collagen-1,matrix metalloproteinase-2(MMP-2)and tissue inhibitors of metalloproteinase-1(TIMP-1),the protein expression of 5-LO were evaluated by real time PCR and Western blot respectively. Histological analysis was performed by microscopy observation. Fibrosis conditions in rat liver in each group were evaluated according to the semi-quantitative scoring system (SSS), Hyp in rat livers and the hepatic functional biochemistry were also detected. Results Along with the aggravation of liver fibrosis, the gene expression of 5-Lo,α-SMA,Collagen-1,MMP-2 and TIMP-1 increased gradually, and the level of ALT, TBA, Hyp and SSS score also increased gradually. With the administration of AA-861,the Mrna expression of 5-LO,α-SMA,Collagen-1,MMP-2,TIMP-1 and the protein expression of 5-LO decreased remarkably, and the reduction in TIMP-1 Mrna expression was more significant than that in MMP-2.Six weeks after AA-861 treatment, the level of ALT, TBA, Hyp and SSS score were also decreased significantly. Conclusion AA-861 ameliorates CCI4 induced rat liver fibrosis by inhibiting the 5-LO pathway and decreasing the expression of 5-LO,α-SMA,Collagen-1,MMP-2 and TIMP-1.

Objective To investigate the distribution characteristic and drug resistance of respiratory tract pathogens in ICU eld‐erly patients to provide the basis for clinical medication and control of nosocomial infection .Methods The isolation situation and drug resistance of pathogens in ICU elderly patients with respiratory tract infection from January 2012 to December 2014 were ret‐rospectively analyzed .Results Among 501 cases of respiratory tract infection ,350 cases were Gram‐negative bacilli infection ,which were mainly P .aeruginosa and A .baumannii;50 cases were Gram‐positive coccus infection ,which was mainly S .aureus ;101 cases were complicating fungal infection ,which was mainly C .albicans .The resistance of P .aeruginosa to imipenem showed upward trend (P<0 .05) ,A .baumannii had higher resistance to commonly used antimicrobial drugs ,but the drug resistance trend had no obvious change(P>0 .05) .Imipenem‐resistant A .bauman(IRAB) ,ESBLs‐producing E .coli and methicillin‐resistant S .aureus (MRSA) in the elderly patients with respiratory tract infection all exceeded 50% of each constitution ratio .Conclusion Multi‐drug resistant bacteria are usually isolated from ICU elderly patients ,their drug resistance rates maintain a higher level .Therefore clinicians should rationally select antibacterial drugs by combining with the laboratory reports ,increase the prevention and management of multi‐drug resistant bacteria and reduce the nosocomial infection occurrence .

Abstract?AIM: To observe the clinical efficacy of intravitreal Lucentis injection combined with panretinal photocoagulation ( PPR ) and compound trabeculectomy for neovascular glaucoma ( NVG) .?METHODS:A total of 14 cases (14 eyes) with NVG were collected from January to November 2015.All cases were treated with intravitreal lucentis injection, PPR and compound trabeculectomy by turns.Intraocular pressure ( IOP) , visual acuity and the complications at pre-or post-surgery were recorded, respectively.?RESULTS: Followed up for 3-6mo, the average IOP preoperatively was significantly decreased than that detected at post-operation ( 18.00 ±6.70 vs 41.65 ± 4.07mmHg, t=11.288, P<0.05).IOP less than 21mmHg with or without the usage of anti-intraocular pressure drugs was defined as the sign of successful or effective surgery, respectively.At the ultimate follow-up, 11 cases were successful, 2 cases were effective, and the success rate was 79%, effective rate was 14%.Only 1 case was applied cyclocryotherapy due to the uncontrolled IOP. Moreover, the results of visual acuity detection demonstrated that 6 eyes got a better visual acuity, 7 eyes remained the same condition and 1 case got no light perception. Meanwhile, 13 cases showed none iris neovascularization during the follow-up; 1 case got a reappearance of iris neovascularization on the third month, which was then dissolved subjected to the intravitreal lucentis injection in combination with PPR. One case developed post -operative hyphema and absorbed after 1wk. No shallow anterior chamber and eyeball atrophy happened.?CONCLUSION:Intravitreal lucentis injection combined with PPR and compound trabeculectomy is an effective and safe therapeutic strategy for the treatment of NVG.

Objective To explore the clinical value of multi-slice CT enhancement perfusion imaging (MSCTPI) in diagnosis and differential diagnosis of thyroid disease. Methods Thirty-three patients with benign thyroid diseases were enrolled in the benign group, and were divided into subgroups of nodular goiter (n=17) and thyroid adenoma (n=16), while 10 patients with thyroid carcinoma were enrolled in the malignant group. All patients underwent routine CT scanning and MSCTPI with GE LightSpeed 16-detector row CT scanner. Time-density curve (TDC) of common carotid for benign thyroid diseases and thyroid carcinoma was depicted. Perfusion parameters of blood flow (BF), blood volume (BV), mean transit time (MTT) and permeability surface area product (PS) were obtained automatically. All parameters were statistically analyzed among groups. Results TDC showed single peak in common carotid artery, with small peak of speed up and slow down in benign group, while with baseline segment, up above, down segment and horizontal segment in thyroid carcinoma. There was statistical difference between benign and malignant groups in BF, BV, MTT and PS value (P=0.001, <0.001, 0.003 and <0.001, respectively). No significant difference of BV and MTT was found between subunits of benign and thyroid carcinoma (all P>0.05). BF was significantly different in benign and maligant groups (P<0.05), whereas PS in thyroid carcinoma and nodular goiter was significant different (P<0.05). No statistical difference of BF, BV, MTT and PS was detected between nodular goiter and thyroid adenoma. Conclusion MSCTPI can exactly show the blood flow features of thyroid. The analysis of BF, BV, MTT and PS is helpful for differential diagnosis between benign thyroid disease and thyroid carcinoma.