Critical Review of Synthesis, Toxicology and Detection of Acyclovir.

Acyclovir (ACV) is an effective and selective antiviral drug, and the study of its toxicology and the use of appropriate detection techniques to control its toxicity at safe levels are extremely important for medicine efforts and human health. This review discusses the mechanism driving ACV's ability to inhibit viral coding, starting from its development and pharmacology. A comprehensive summary of the existing preparation methods and synthetic materials, such as 5-aminoimidazole-4-carboxamide, guanine and its derivatives, and other purine derivatives, is presented to elucidate the preparation of ACV in detail. In addition, it presents valuable analytical procedures for the toxicological studies of ACV, which are essential for human use and dosing. Analytical methods, including spectrophotometry, high performance liquid chromatography (HPLC), liquid chromatography/tandem mass spectrometry (LC-MS/MS), electrochemical sensors, molecularly imprinted polymers (MIPs), and flow injection-chemiluminescence (FI-CL) are also highlighted. A brief description of the characteristics of each of these methods is also presented. Finally, insight is provided for the development of ACV to drive further innovation of ACV in pharmaceutical applications. This review provides a comprehensive summary of the past life and future challenges of ACV.

Menatetrenone versus alfacalcidol in the treatment of Chinese postmenopausal women with osteoporosis: a multicenter, randomized, double-blinded, double-dummy, positive drug-controlled clinical trial.

OBJECTIVE: To evaluate whether the efficacy and safety of menatetrenone for the treatment of osteoporosis is noninferior to alfacalcidol in Chinese postmenopausal women. METHOD: This multicenter, randomized, double-blinded, double-dummy, noninferiority, positive drug-controlled clinical trial was conducted in five Chinese sites. Eligible Chinese women with postmenopausal osteoporosis (N=236) were randomized to Group M or Group A and received menatetrenone 45 mg/day or alfacalcidol 0.5 µg/day, respectively, for 1 year. Additionally, all patients received calcium 500 mg/day. Posttreatment bone mineral density (BMD), new fracture onsets, and serum osteocalcin (OC) and undercarboxylated OC (ucOC) levels were compared with the baseline value in patients of both groups. RESULTS: A total of 213 patients (90.3%) completed the study. After 1 year of treatment, BMD among patients in Group M significantly increased from baseline by 1.2% and 2.7% at the lumbar spine and trochanter, respectively (P<0.001); and the percentage increase of BMD in Group A was 2.2% and 1.8%, respectively (P<0.001). No difference was observed between groups. There were no changes in femoral neck BMD in both groups. Two patients (1.9%, 2/108) in Group M and four patients (3.8%, 4/105) in Group A had new fracture onsets (P>0.05). In Group M, OC and ucOC decreased from baseline by 38.7% and 82.3%, respectively (P<0.001). In Group A, OC and ucOC decreased by 25.8% and 34.8%, respectively (P<0.001). Decreases in serum OC and ucOC were more obvious in Group M than in Group A (P<0.001). The safety profile of menatetrenone was similar to alfacalcidol. CONCLUSION: Menatetrenone is an effective and safe choice in the treatment of postmenopausal osteoporosis in Chinese women.

[Survey on characteristics of menopause of Chinese women with the age of 40-60 years at gynecological clinic from 14 hospitals].

OBJECTIVE: To investigate the characteristics of menopause of Chinese women with the age of 40-60 years concerning gynecologic clinics in China. METHODS: From Mar.2008 to Sept.2008, a face-to-face questionnaire was conducted in gynecological clinic in perimenopausal and postmenopausal women in 14 hospitals in China, which included general demographic data, menstrual change process, climacteric symptoms and knowledge about menopause. Modified Kupperman index were used to evaluate climacteric symptoms during the recent week and awareness of hormonal replacement therapy were studied. RESULTS: A total of 1641 women were investigated. The ages of onset of menopause transition, climacteric symptoms and natural menopause were (47 ± 4), (46 ± 4), (49 ± 3) years old respectively. Climacteric symptoms could be found in 78.43% (1287/1641) women during menopausal transition, which were mainly mild to moderate symptoms. The top 5 symptoms were fatigue and weakness (71.48%, 1173/1641), irritability (68.68%, 1127/1641), insomnia (67.65%, 1110/1641), muscle and joint pain (64.11%, 1052/1641) and hot flush (57.90%, 950/1641). The climacteric symptoms were not constant during menopausal transition, usually more severe in late transition and postmenopausal periods, during which the moderate and severe symptoms were 59.1% (189/320) and 51.1% (291/570) respectively. Although most symptoms primarily appeared along with menstruation change, there are about 17.5% (172/981) patients experienced climacteric symptoms before menstruation change occurrence. There were 56.39% (733/1300) women had ever heard (mostly from gynecologist) about hormone replacement therapy from Obstetrician and Gynecologist. CONCLUSIONS: Most of the women during menopausal transition had climacteric symptoms, usually mild and moderate ones. Although most symptoms primarily appeared along with menstruation change, there are other patients' experienced climacteric symptoms before menstruation change occurrence.

Efficacy and safety of 2 g/day of strontium ranelate in Asian women with postmenopausal osteoporosis.

Strontium ranelate is a new effective anti-osteoporotic treatment having a unique mode of action, reducing bone resorption while promoting continued bone formation, with a broad range of anti-fracture efficacy at vertebral as well as peripheral sites. In Phase III studies, it has proven its early and sustained efficacy against vertebral fractures in Caucasians along with a significant increase in lumbar bone mineral density (BMD). The aim of this randomized double-blind study was to demonstrate the efficacy of strontium ranelate (2 g/day) on lumbar spine bone mineral density and the clinical and biological safety in Asian postmenopausal osteoporotic patients compared to placebo over 1 year. Three hundred and twenty-nine eligible women from mainland China, Hong Kong and Malaysia were randomized into the study. The baseline characteristics were similar in the treatment and placebo groups: mean age of 66.2+/-6.5 years, time since menopause 17.6+/-7.2 years. In the Full Analysis Set (FAS, N=302), the mean baseline lumbar L2-L4 BMD was 0.715+/-0.106 g/cm(2) in the strontium ranelate group and 0.708 +/- 0.109 g/cm2 in the placebo group. The mean baseline femoral neck BMD was 0.575+/-0.074 g/cm2 and 0.566+/-0.069 g/cm2 respectively and mean total hip BMD was 0.642+/-0.080 g/cm2 and 0.631 +/-0.088 g/cm2 respectively. The overall compliance was 91.4% in the study drug group, and 97.4% in the placebo group. After 1 year of treatment, the lumbar spine, femoral neck and total hip BMD in the treated group was significantly increased by 3-5% as compared to placebo. Strontium ranelate was well tolerated. The most frequently reported emergent adverse events were comparable in both groups (60.4% versus 60.0%), with majority of them being mild gastrointestinal disorders. There were no clinically relevant changes in laboratory tests, such as blood routine, hepatic and renal function. It is thus concluded that the effects of 2 g/day strontium ranelate on BMD and its safety profile in this cohort of postmenopausal osteoporotic Asian women were consistent with results obtained from Caucasian women in which the efficacy on the reduction in risk of fracture has been proven.

On the sulfation and methylation of catecholestrogens in human mammary epithelial cells and breast cancer cells.

Prolonged exposure to high level of estrogen is a known risk factor for breast carcinogenesis. In cells, estrogens, in particular estrone (E1) and 17 beta-estradiol (E2), can be converted to catecholestrogens (CEs) which may be oxidized to form CE-semiquinones and CE-quinones that are capable of binding to DNA to induce mutations, followed by carcinogenesis. Whether the body is equipped with protective mechanisms against potentially harmful CEs, therefore, is an important issue. The present study was designed to examine the role of sulfation in the metabolism of CEs. MCF-7 breast cancer cells and MCF 10A human mammary epithelial cells were metabolically labeled with [35S]sulfate in the presence of individual CEs. Analysis of the labeling media showed the generation and release of exclusively [35S]sulfated 2-methoxy-E1 or [35S]sulfated 2- or 4-methoxy-E2 by cells labeled in the presence of 2-OH-E1 or 2- or 4-OH-E2. Whereas both [35S]sulfated 4-methoxy-E1 and [35S]sulfated 4-OH-E1 were detected in the labeling media of cells labeled in the presence of 4-OH-E1. These results indicated a concerted action of catechol-O-methyltransferase (COMT) and the cytosolic sulfotransferase (SULT) enzyme(s) in the metabolism of CEs. Enzymatic assays revealed that, five (SULT1A1, SULT1A2, SULT1A3, SULT1C4, and SULT1E1) of eleven known human SULTs tested could use CEs and methoxyestrogens (MEs) as substrates, with SULT1E1 displaying the strongest sulfating activity.

[Expression of estrogen sulfotransferase (EST) gene and bridging integrator protein-1 (BIN1) gene and their significance in breast tissues].

BACKGROUND & OBJECTIVE: Estrogen sulfotransferase (EST) is an enzyme which metabolizes estrogen into inactive estrogen sulphate. Bridging integrator protein-1 (BIN1) is a novel c-myc-interacting protein with the features of tumor suppressor. EST and BIN1 may be protective in tumorigenesis. This study was to detect the gene expression of EST and BIN1 in breast cancer tissues,and further investigate the carcinogenesis mechanism of breast cancer. METHODS: The mRNA levels of EST and BIN1 in 12 specimens of normal human breast tissue, and 24 specimens of breast cancer were measured by reverse transcriptase-polymerase chain reaction (RT-PCR). RESULTS: EST mRNA and BIN1 mRNA were expressed in all normal human breast tissues, while EST mRNA was decreased in 75.0% (18/24), and lost in 25.0% (6/24) breast cancer specimens; BIN1 mRNA was decreased in 25.0%(16/24), and lost in 66.7.0% (16/24) breast cancer specimens. CONCLUSION: Down-regulation of EST mRNA and BIN1 mRNA may play an important role in the molecular mechanisms of breast carcinogenesis.

[Effects of phytoestrogen alpha-zearalanol on normal human breast].

OBJECTIVE: To study the effects of phytoestrogen alpha-zearalanol (alpha-ZAL) on normal human breast. METHODS: Ten specimens of normal human breast tissues were subcutaneously implanted into 30 athymic nude mice aged 9-10 weeks, one for 3 mice. These mice were then randomly divided into three groups: control group (without hormone treatment, n = 10), 1 mg/kg alpha-ZAL group (n = 10), and 5 mg/kg alpha-ZAL group (n = 10). All breast tissues were taken out 6 weeks later. Immunohistochemistry was used to determine the protein expressions of proliferating cell nuclear antigen (PCNA), inhibiting apoptosis gene Bcl-2, estrogen receptor (ER), and progesterone receptor (PR). Reverse transcription polymerase chain reaction (RT-PCR) was used to measure the expression levels of estrogen sulfotransferase (EST) mRNA and bridging integrator protein-1 (BIN1) mRNA. Morphological features of grafts before and after treatment were also observed. RESULTS: Alpha-ZAL had no significant effects on Bcl-2, PCNA, ER, and PR expression of mammary epithelial cells in graft specimens. Alpha-ZAL upregulated BIN1 mRNA expression in grafts, but had no significant effect on ESTmRNA expression. CONCLUSIONS: Alpha-ZAL does not affect the morphology, proliferating, and apoptosis of epithelial cells in normal human breast tissues implanted into nude mice, but it may increase the gene expression of tumor-inhibiting BIN1, suggesting that alpha-ZAL may have potential proteotive effect on normal human breast.

[Comparison of the effect of fluoxetine combined with hormone replacement therapy (HRT) and single HRT in treating menopausal depression].

OBJECTIVE: To compare the treatment effects between fluoxetine plus hormone replacement therapy (HRT) and HRT alone on menopausal depression. METHODS: A total of 54 women with climacteric symptoms and depression were enrolled and randomly divided into two groups. The fluoxetine plus HRT group received fluoxetine (20 mg, qd, po) and cyclic use of conjugated estrogen 0.625 mg and medroxyprogesterone acetate 5 mg, while HRT group was assigned to receive cyclic use of conjugated estrogen 0.625 mg and medroxyprogesterone acetate 5 mg only. All subjects were interviewed and evaluated with Hamilton Depression score (HAMD) and Kupperman menopause index (KMI) at week 0, 1, 2, 3, 4, 6 and 8 of the treatment. RESULTS: There was no difference between two groups in HAMD and KMI before the trial (P > 0.05). HAMD scores at the 8th week in fluoxetine plus HRT group and HRT group were 3.0 +/- 2.3 and 11.2 +/- 5.8 respectively, being significantly different (P < 0.001). At the 6th week of treatment KMIs in fluoxetine plus HRT group and HRT group were 9 +/- 6 and 14 +/- 7 respectively, also significantly different (P < 0.05). The healing rates of fluoxetine plus HRT group and HRT group were 92% and 48% respectively, which were statistically different by Chi square test (P < 0.001). CONCLUSIONS: The effect of fluoxetine plus HRT on menopausal depression is significantly superior than that of HRT only and the difference becomes more obvious with treatment time. Both groups could ameliorate the climacteric symptoms with the effect of fluoxetine plus HRT more obvious.

Menopausal depression: comparison of hormone replacement therapy and hormone replacement therapy plus fluoxetine.

BACKGROUND: To compare the efficacy and safety of hormone replacement therapy (HRT) combined with fluoxetine, with HRT alone, in post-menopausal women suffering from depression. METHODS: A randomized, open-label, parallel trial was applied. HRT was administered to all patients for 2 cycles, with 14 days of estrogen therapy and 14 days of estrogen plus progesterone. Patients who were randomly assigned to the HRT plus fluoxetine group were given fluoxetine in combination with HRT. Hamilton Depression Rating Scale (HAMD), Kupperman Menopausal Index (KMI), and Clinical Global Impressions scale were used to measure the efficacy. RESULTS: One hundred and twenty-three post-menopausal patients with depression were enrolled in the study. Among them, 120 had at least one post-treatment visit and entered into the statistical analysis. The mean total HAMD scores were significantly lower, and the percentages of HAMD score reductions were higher in the HRT plus fluoxetine Group compared with the HRT Group, after at least 3 weeks of treatment, with an average difference of 5 points at the endpoint. The Clinical Global Impression-Severity and Clinical Global Impression-Improvement scores were significantly different in the 2 groups, in favor of the combination therapy. The mean total KMI was significantly lower in the Combination Group compared with the HRT Group, after at least 6 weeks of treatment, with an average 4.5-point difference between the groups. No statistically significant differences were found in most of the adverse events reported in the Combination Group compared with the HRT group, with the exception of 3 symptoms, i.e., dry mouth, loss of appetite, and abdominal distention. They were mild to moderate in severity. Two patients in the HRT group, but none in the combination group, dropped out due to adverse events. CONCLUSION: HRT plus fluoxetine therapy was effective in the treatment of menopausal depression with a satisfactory safety profile.

[Expression of estrogen sulfotransferase in breast tissues and the regulation of ESTmRNA by estrogen].

OBJECTIVE: To detect the expression of estrogen sulfotransferase (EST) in breast tissues, and to investigate its significance in breast carcinogenesis and the relationship between EST mRNA and estrogen. METHODS: Seven specimens of normal human breast tissues from patients with benign breast diseases were implanted subcutaneously into athymic nude mice, one specimen divided into many pieces to be implanted into 2 groups of mice, totalling 14 mice. Then the 14 mice were divided into 2 groups: as control and the other injected intramuscularly with estradiol benzoate (B-E(2)) every other day for 6 weeks. Six weeks after, the implanted tissues were taken out. Reverse transcription polymerase chain reaction (RT-PCR) was used to detect the expression of EST mRNA in 24 specimens of breast cancer, 6 specimens of normal breast tissues around the breast fibroadenoma, all resected during operation, and the specimens of implanted normal breast tissues from the mice (control group and B-E(2) group). The protein expression of estrogen receptor (ER) was detected by immunohistochemistry in the 24 specimens of breast cancer. RESULTS: EST mRNA was expressed in all normal human breast tissues and the expression was increased in the grafts from mice of B-E(2) group. However, the specimens of breast cancer showed decrease or absence of EST mRNA expression. CONCLUSION: EST mRNA expression is decreased or lost in breast cancer tissues. EST mRNA can be regulated by estrogen in normal breast tissues.

[The effect of estrogen benzoate on normal human breast tissue implanted into athymic nude mice].

OBJECTIVE: To investigate the proliferating effect of estradiol benzoate on normal human breast tissue. METHODS: Three specimens of normal human breast tissue were implanted into nine 9-10-week-old intact female athymic nude mice which were randomly divided into group A, B and C. Each specimen was divided into three parts, and each part was implanted into mice of each group of three (A, B and C) respectively. Two weeks later, part of xenografts was taken out from group A as control group, and then group A, B and C were injected muscularly with 3, 6, 9 micrograms estradiol benzoate once daily, respectively. After therapy for 28 days, the grafts were taken out and expression of proliferating cell nuclear antigen (PCNA), estrogen receptor (ER) and proliferating receptor (PR) in grafts were examined using immunohistochemical method. RESULTS: Compared with control group, PCNA was increased in group B and C (P < 0.05), but in group A had no significant change. ER was gradually down regulated by 3, 6 and 9 micrograms estradiol benzoate (P < 0.05); however, PR was increasingly up regulated (P < 0.05). CONCLUSIONS: Certain dosage of estradiol benzoate has proliferating effect on normal human breast tissue.

[A randomised clinical trial to study the effects of raloxifene hydrochloride on bone mineral density, biochemical markers of bone metabolism and serum lipids in postmenopausal women].

OBJECTIVE: To determine the effect of raloxifene hydrochloride (RLX) on bone mineral density (BMD), biochemical markers of bone metabolism and lipid metabolism in Chinese postmenopausal women. METHODS: This was a multicenter, randomized, double blind placebo controlled study in China with a total of 204 postmenopausal women [mean age (60 +/- 5) years (x +/- s) and weight (63 +/- 9) kg (x +/- s)] treated with either RLX 60 mg (n = 102) or placebo (n = 102) daily for 12 months. BMD, serum lipid and bone markers were determined before and after drug administration. RESULTS: Compared to placebo, RLX produced a significant increase in both total lumbar spine and total hip BMD. For the lumbar spine, percentage increase in total BMD was 2.30% with RLX compared to a decrease of 0.08% with placebo (P < 0.001). Corresponding values for total hip BMD were 2.46% increase for RLX and 1.07% for placebo (P < 0.05). For biochemical markers of bone metabolism, serum osteocalcin and C-telopeptide, percentage decrease were 27.6% and 24.0% in raloxifene-treated subjects. Corresponding values in placebo were 10.6% decrease and 15.8% increase (RLX compared to placebo, both P < 0.001). For total cholesterol and low-density lipoprotein cholesterol, percentage decrease were 6.4% and 34.6% in the raloxifene-treated group. Corresponding values in placebo were 1.4% increase and 19.1% decrease (RLX compared to placebo, both P < 0.001). No differences were found for high-density lipoprotein cholesterol or triglyceride levels between the two groups. Only 5 subjects discontinued early due to an adverse event (3 in the RLX group and 2 in the placebo group). CONCLUSIONS: This study confirms that RLX exerts positive effects on the skeleton, increasing BMD and decreasing biochemical markers of bone metabolism, and decreased total cholesterol and low-density lipoprotein cholesterol in postmenopausal women in China.