0. 05) , while (0. 62?0. 09)mmol/g in the control group. Both ambroxol and dexamethasone group had significant difference with the control group (P

Objective To evaluate clinical efficacy of multiple regimen combination in treatment of osteoporosis of perimenopausal or postmenopausal women. Methods From Jul. 2008 to Dec. 2009, 109 women with low bone mineral density (BMD) or osteoporosis treated in Department of Obstetrics and Gynecology, Affiliated Second Hospital, Wenzhou Medical College were enrolled randomly into 3 group,including 36 women in Group A managed by osteoform 1000 mg/d + alfacalcidol 0. 25 μg/bid orally, 40 women in group B managed by osteoform 1000 mg/d + alfacalcidol 0. 25 μg/bid + tibolone 1.25 mg/d orally and 33 women in group C managed by ostcoform 1000 mg/d + alfacalcidol 0. 25 μg/bid +bisphosphonates 70 mg/w orally. After 48 weeks BMD on lumbar 1 -4 (L1-4) and left femur were detected by X-ray. Bone alkaline phosphatase(BALP) ,cross linked clelopeptide of type Ⅰ collagen(CTX) and 25-hydroxychole calciferol [25 (OH) D3] was measured by enzyme linked immunosorbent assay (ELISA).Result Seven women (6. 4%, 7/109) were withdrawed form this study, including 2 cases losing follow up in group A, 3 cases stopping treatment in group B, 2 cases giving up treatment due to severe adverse effect (burning in upper abdomen) in group C. (1) Pain relieve: after 48 weeks treatment, women in 3 groups improved symptom of pain significantly, the rates of pain relieve were 85% (29/34)in group A, 92% (34/37) in group B and 94% (29/31) in group C. (2) BMD: BMD was improved significantly in women in 3 groups after treatment. BMD of L1-4 were (0.88±0.15) g/cm2 in group A,(0.89±0.18) g/cm2 in group B and (0.87±0.10) g/cm2 in group C before treatment, and converted to (0.90±0.01) g/cm2 in group A, (0.93±0.09) g/cm2 in group B and (0.91±0.11) g/cm2 in group C after treatment. BMD of left femur were (0.87±0.07) g/cm2 in group A, (0.87±0.07) g/cm2 in group B and (0.85±0. 12) g/cm2 in group C before treatment and converted to (0.90± 0.03) g/cm2 in group A, (0.91±0.08) g/cm2 in group B and (0.89 ±0.12) g/cm2 in group C after treatment. It was shown significantly different BMD between group B or C and group A (P < 0. 01), however, there was no significant different BMD between group B and C (P >0. 05). (3) Index of bone metabolism: BALP were (26±6) μg/L in group A, (26±9) μg/L in group B and (28±7) μg/L in group C before treatment and converted to (22±5) μg/L in group A, (20±9)μg/L in group B and (22±8)μg/L in group C after treatment, which showed statistical difference (P < 0.05). CTX were (0.85±0.20) ng/L in group A, (0.84±0.47) ng/L in group B, and (0. 88 ±0. 11) ng/L in group C before treatment and converted to (0. 81 ±0. 19) ng/L in group A, (0. 77±0.33) ng/L in group B, and (0.82 ±0. 14) ng/L in group C after treatment, which showed statistical difference (P < 0. 05). Conclusions Those 3 regimens combination could be used in treatment of osteoporosis by decreasing bone conversion, increasing bone density, decreasing bone absorption. Regimen A was only suitable for basic therapy,the other two regimens could provide better treatment.

Objective:To investigate the role of Akt signaling pathway in the regulation of breast cancer bone metastasis induced pain behavior in rat.Methods:Model rats were randomly divided into three groups:Model group,Model+GSK690693 group and Model+ Saline group.On PID 13,14 and 21,Model+GSK690693 group rats were intrathecally injected with GSK690693,a specific inhibitor of Akt.Model+Saline group were injected with saline instead.The pain related behaviors were respectively recorded on PID 0,7,14 and 21.The expression ofp-Akt in DRG used for western bloting were examed on PID 21.Results:After the injection of Akt specific inhibitor GSK690693 by intrathecal,In the Model+GSK690693 group,the threshold value of mechanical contraction reflex was increased,the spontaneous pain behavior and the expression of p-Akt in DRG decreased.On PID 14 d and 21 d,the pain behavior of rats in Model+GSK690693 group was significantly different from that of Model+Saline group and Model group (P＜0.01);On PID 21 d,There was significant statistical significance (P＜0.01) on the expression of p-Akt and Model in the ipsilateral Model+GSK690693 of DRG group,which was statistically significant (P＜0.05) compared with the Model+Saline group.Conclusion:Intrathecal injection of Akt specific inhibitor GSK690693 inhibits rat pain related behaviors induced by bone metastasis in rat breast cancer.

OBJECTIVE: To evaluate the efficacy and safety of thread-dragging through fistula method in treating patients with simple anorectal fistula. METHODS: In this multi-centered, prospective, and randomized controlled clinical trial, 244 patients with simple low or high anorectal fistula were randomly divided into study group (with the method of thread-dragging through fistula) and control group (with the method of incision or thread-drawing). The healing time and curative rate of anorectal fistula, and the integral calculus of clinical symptom and life quality evaluations before and after treatment were all examined. The maximal anal canal squeeze pressure was measured to compare the therapeutic safety between these two groups. The health economical benefits were also assessed to determine which therapeutic method was more economical. RESULTS: The curative rate of simple low and high anorectal fistula were of no significant differences between the study group and the control group. The healing time of simple low anorectal fistula in the study group and the control group were (22.26+/-8.67) d and (31.41+/-11.39) d respectively, while the healing time of simple high anorectal fistula in the study group and the control group were (24.73+/-8.15) d and (32.20+/-12.60) d respectively, and there revealed significant differences between these two groups. Each integral calculus of clinical symptom evaluation in the study group was not obviously different from those in the control group besides the integral calculus of anal sphincter function. The integral calculus of life quality between the study group and the control group of simple low anorectal fistula had no significant differences. The integral calculus of anal sphincter function and confidence in treatment in the study group of high anorectal fistula were better than those in the control group. The hospitalization expense of the study group was remarkably lower than that of the control group. The maximal anal canal squeeze pressure in the study group after treatment was not reduced obviously as compared with that in the same group before treatment, while it was decreased significantly in the control group after treatment as compared with those in the same group before treatment and in the study group after treatment. CONCLUSION: The method of thread-dragging through fistula in treating simple low and high anorectal fistula can shorten the course of the disease, save the hospitalization expenses, improve the life quality of the patients, and protect the anal sphincter function.

OBJECTIVE: To analyze the phenotype and genetic variant in a pedigree affected with inherited protein C (PC) deficiency. METHODS: The proband and her family members (7 individuals from 3 generations) were tested for plasma protein C activity (PC:A), protein C antigen (PC:Ag) content and other coagulation indicators. All of the 9 exons and flanking sequences of the proband's PROC gene were amplified by PCR and sequenced. Suspected variants were verified by reverse sequencing of the proband and her family members. Bioinformatic software was used to analyze the pathogenicity and conservation of the variant site. Swiss-PdbViewer was used to analyze the three-dimensional model and the interaction with the mutant amino acid. RESULTS: The PC:A and PC:Ag of the proband, her grandmother, father and elder brother were decreased to 55%, 52%, 48%, 51% and 53%, 55%, 50%, 56%, respectively. Genetic analysis showed that the four individuals have all carried heterozygous c.1318C>T (p.Arg398Cys) missense mutation in exon 9 of the PROC gene. The score of MutationTaster was 0.991, PROVEAN was -3.72, and FATHMM was -2.49, all predicted it to be a harmful mutation. Phylogenetic analysis also showed that Arg398 was weakly conservative among homologous species. Protein model analysis showed that, in the wild type, Arg398 can form a hydrogen bond with Glu341 and Lys395 respectively, when it was mutated to Cys398, the hydrogen bond with Glu341 disappears and an additional hydrogen bond was formed with Lys395, which has changed the spatial structure of the protein. CONCLUSION The heterozygous missense mutation c.1318C>T (p.Arg398Cys) of the PROC gene probably underlay the decreased PC:A and PC:Ag in this pedigree.
Aged ; Female ; Heterozygote ; Humans ; Male ; Mutation ; Pedigree ; Phenotype ; Phylogeny ; Protein C Deficiency/genetics*

OBJECTIVETo identify the genetic mutation underlying congenital hypofibrinogenamia in a Chinese pedigree.

METHODSStandard coagulation tests including the prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), plasminogen activity (PLG:A), D-Dimer (DD) and fibrin degradation products (FDP) were tested with fresh plasma using a STA-R analyzer. The activity of fibrinogen (Fg:C) and fibrinogen antigen (Fg:Ag) were measured respectively with the Clauss method and immunoturbidimetry. All exons and exon-intron boundaries of the fibrinogen Aα-, Bβ-, and γ-chain genes (FGA, FGB and FGG) were amplified by PCR followed by direct sequencing. Suspected mutation was confirmed by reverse sequencing and analyzed with a Swiss-PdbViewer.

RESULTSThe PT level in the proband was normal, while the APTT and TT were slightly prolonged. The functional and antigen fibrinogen levels were both significantly reduced (0.91 g/L and 0.95 g/L, respectively). Similar abnormalities were also found in her father, elder sister, son and niece. The coagulant parameters of her mother were all within the normal range. Genetic analysis has reveled a heterozygous A>C change at nucleotide 5864 in exon 7 of γ gene in the proband, predicting a novel Lys232Thr mutation. The proband's father, elder sister, son and niece were all carriers of the same mutation. Protein model analysis indicated that the Lys232Thr mutation did not disrupt the native network of hydrogen bonds, but has changed the mutual electrostatic forces, resulting in increased instability of the protein.

CONCLUSIONThe heterozygous Lys232Thr mutation identified in the FGG gene probably underlies the hypofibrinogenemia in this pedigree.

Adult ; Afibrinogenemia ; congenital ; genetics ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; China ; Female ; Fibrinogen ; genetics ; Humans ; Male ; Middle Aged ; Molecular Sequence Data ; Pedigree ; Peptide Fragments ; genetics ; Young Adult

OBJECTIVETo explore the molecular pathogenesis and clinical phenotypes in 10 probands with inherited fibrinogen (Fg) deficiency.

METHODSThe diagnosis of hereditary Fg deficiency was validated by prothrombin time (PT), thrombin time (TT), Fg activity (Fg:C) and Fg antigen (Fg:Ag) in plasma. All of the exons and their flanking sequences of the Fg gene were analyzed by direct sequencing. Detected mutations were confirmed by reverse sequencing.

RESULTSThe ranges of Fg:C and Fg:Ag in the 10 probands were 0.52-0.91 g/L and 0.62-2.98 g/L, respectively. Five of the probands had type I disorders, and 5 had type II disorders. Seven point mutations were identified, among which 6 have located in the D region. γThr277Arg, γAsp316His, γTrp208Leu and Lys232Thr were novel mutations, and αArg19Ser was first reported in Chinese. Four probands had the same mutation site (γArg275). As to the clinical manifestation, probands with type I disorders were asymptomatic or with mild or medium symptoms, while those belonged to type II disorders had moderate or serious symptoms. Two probands have carried an Arg275Cys mutation but had different clinical manifestations.

CONCLUSIONMutations of the Fg gene seem to aggregate to the D region of FGG in our region, and Arg275 is a common mutation. However, no correlation has been found between the mutation site and clinical manifestations.

Adolescent ; Adult ; Afibrinogenemia ; blood ; classification ; genetics ; Base Sequence ; Child ; DNA Mutational Analysis ; methods ; Exons ; genetics ; Family Health ; Female ; Fibrinogen ; genetics ; metabolism ; Genotype ; Humans ; Male ; Middle Aged ; Mutation, Missense ; Partial Thromboplastin Time ; Phenotype ; Point Mutation ; Polymerase Chain Reaction ; Prothrombin Time ; Thrombin Time ; Young Adult

Objective:To explore the changes in serum levels of miR-499 and miR-362 in patients with advanced non-small cell lung cancer (NSCLC) and their relationship with prognosis.Methods:A total of 103 patients with advanced NSCLC at Shanghai University of Medicine & Health Sciences Affiliated Chongming Hospital from January 2020 to October 2021 were selected as the NSCLC group, and 100 healthy volunteers who underwent physical examinations at our hospital during the same period were selected as the control group. Fluorescent quantitative PCR was used to determine and compare the levels of serum miR-499 and miR-362 in the two groups, and the relationship between the two indexes and different clinical characteristics of NSCLC patients was analyzed. According to the clinical outcome of 2-year follow-up, the patients were divided into survival group and death group, and the levels of serum miR-499 and miR-362 were compared between the two groups. The predictive value of miR-499 and miR-362 levels on the prognosis of advanced NSCLC patients were analyzed using receiver operator characteristic (ROC) curves.Results:The serum miR-499 level in the NSCLC group (0.34±0.10) was lower than that in the control group (1.25±0.21), while the miR-362 level (1.13±0.27) was higher than that in the control group (0.63±0.15) ( t=18.26, P<0.001; t=16.32, P<0.001). There were statistically significant differences in serum miR-499 and miR-362 levels among patients with different degrees of differentiation ( t=11.12, P<0.001; t=16.35, P<0.001), TNM staging ( t=13.64, P=0.002; t=8.73, P=0.010) and lymph node metastasis ( t=10.02, P=0.003; t=9.65, P=0.004). The serum miR-499 level in the death group ( n=77) (0.24±0.06) was lower than that in the survival group ( n=26) (0.35±0.09), while the miR-362 level (1.54±0.32) was higher than that in the survival group (1.08±0.21), with statistically significant differences ( t=8.06, P=0.006; t=8.67, P=0.005). ROC curve analysis showed that the sensitivity of miR-499 and miR-362 in predicting the prognosis of advanced NSCLC patients was 73.46% and 75.85%, respectively, with specificity of 64.42% and 65.61%, AUC of 0.739 (95% CI: 0.662-0.805) and 0.743 (95% CI: 0.640-0.793) ; the sensitivity, specificity, and AUC of serum miR-499 combined with miR-362 in predicting the prognosis of advanced NSCLC patients were 87.63%, 85.34%, and 0.875 (95% CI: 0.698-0.897), respectively; the combined prediction of miR-499 and miR-362 for AUC area was higher than the individual prediction ( Z=4.83, P=0.013; Z=5.17, P=0.009) . Conclusion:Advanced NSCLC patients show significant abnormal serum level of miR-499 and miR-362, and as the severity of the disease progressed, the serum level of miR-499 is downregulated more significantly and miR-362 is upregulated more significantly. The combined detection of miR-499 and miR-362 levels has certain predictive value for the prognosis of advanced NSCLC patients.

Objective:To explore the clinical effect of ALTF transfer with cross-limb bridged "Y" shape vascular anastomosis in repair of complex soft tissue defects in calf.Methods:From August 2010 to September 2020, 33 patients(23 males and 10 females) with complex traumatic soft tissue defects in calf were treated. Preoperative angiography and intraoperative exploration confirmed that there was only 1 main vessel remained in the affected calf or the wound surface and the vessel could not be anastomosed with the vascular pedicle of the ALTF. The size of flap were 16 cm × 8 cm to 25 cm × 18 cm. Two patients received bilobed ALTFs. A "Y" shape anastomosis between the artery of ALTF vascular pedicle and the posterior tibial artery of the contralateral lower limbs was made in all 33 patients to establish the blood supply to the transferred free ALTF. The "Y" shape cross-limb bridged blood vessels at the proximal end of the vascular pedicle artery of the flap were embedded at both ends of the cut-off superior ankle posterior tibial artery of the contralateral lower limb. The vein of the flap was anastomosed with the saphenous vein that associates with the posterior tibial artery. The surface of the suspended blood vessel "bridge" was wrapped with a free skin craft, and the lower limbs were fixed in a straight and parallel position with an external fixation frame. The perfused area of the flap was directly sutured or covered with a free skin craft. The vascular bridge was kept for 3 to 6 weeks before being separated. Outpatient follow-up after discharge.Results:All the patients were entered the postoperative follow-up was 13 months to 7 years, in an average of 25 months. Among the 33 flaps, 31 survived completely, except 1 had necrosis and the other 1 had partial necrosis at the distal end of the flap. The flaps received good blood supply, hence with soft texture and satisfactory appearance. Doppler or DSA was performed after the surgery on the posterior tibial artery of the healthy limb, and the vascular pulsation and patency were found normal. Donor sites for the free skin graft healed well.Conclusion:The ALTF transfer with cross-limb bridged "Y" vascular anastomosis is one of the effective techniques and it was employed in the repair of complex defects of calf soft tissue. It solved the tissue that there was only 1 main vessel or even without a suitable vessel could be anastomosed with the pedicle of the flap.